BLD 434 Completed Learning Objectives
BLD 434 Completed Learning Objectives BLD 434
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This 158 page Study Guide was uploaded by Jacob Decker on Sunday October 11, 2015. The Study Guide belongs to BLD 434 at Michigan State University taught by Kathleen Hoag in Fall 2015. Since its upload, it has received 302 views. For similar materials see in Biomedical Sciences at Michigan State University.
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tuesday schedule have any questions about new material talk about it 0 old material overview 0 practice exams overview 0 go through all questions POLL httpwwwmayomedicallaboratoriescomtest catalogClinicalandlnterpretivel61480 Case Discussion Question Set 2 1 Interpret the test results your team received Do you require any confirmatory tests Please ask for the results of those tests now Use the same team order sheet you used on case day 1 2 After reviewing all of the laboratory results are you able to make a definitive diagnosis What is the most likely diagnosis BLD 434 Case 4 SS15 Patient Background Elena Johns is a 20year old student attending the University of Michigan Elena is a sophomore working on her degree in Art amp Design She recently visited the campus student health clinic because of muscle stiffness and soreness sore throat low grade fever and extreme fatigue She has not felt well for several weeks She has also noticed some recent weight loss but attributes this to not having enough energy to cook for herself recently She s missed more classes than usual because she can t seem to get out of bed in the morning Although her symptoms were tolerable she recently developed a rash on her stomach and upper arms that finally prompted the visit to the clinic The examining physician asked Elena about her major her hobbies and her daily activities She was also asked about her medical history whether she had any tattoos and whether she used illicit drugs of any kind Elena indicated that she lives in the dorms on campus and that she normally enjoys doing a lot of outdoor activities especially hiking in the State Park system trails Elena commented that her W when she is outside walking across campus especially The only prescribed medication she takes on a regular basis is oral contraceptives When asked directly about her sexual activities Elena stated that she is intimate with her boyfriend and that they do not use condoms during intercourse since she is on oral contraceptives She has one tattoo that she got during the first semester of her freshmen year to celebrate her independence and being liberated from her overbearing parents This helped us with Lyme and with SLE Syph Confirmatory Test FTAABS Fluorescent Treponemal Antibody Absorption Test Lwnm SLE because of the lower WBC Confirmatory Test FANA followed by ENA WBC x10A3mircoliter RR 45115 182 Neutrophils RR 5070 44 Lymphocytes RR 1842 45 Atypical lymphocytes noted 30 of lymphocytes Case Discussion Question Set 1 1 Given Elena s physical symptoms and history list some possible diseases she could have and why you are considering each one Secondary Syphilis fatigue rash loss of appetite Syphilitic aseptic meningitis SLE pg 225 symptoms RULED OUT Lyme disease RULED OUT Mononucleosis POSITIVE 2 For case days 1 and 2 combined you will be allowed to choose a maximum of 6 laboratory tests from the following list that you can order on Elena Circle the tests that you want performed ordered on day 1 When you have team consensus on which tests you would like to order bring your team sheet to a facilitator to receive the laboratory test results You will be allowed to choose additional confirmatory testing to be done at any time as long as you do not exceed six total laboratory tests orderedreceived Pay attention to which tests are screening tests and which are confirmatory and ask for them in the appropriate order IMPORTANT Keep in mind that you can only order up to 6 tests total including confirmatory tests that you will be able to request on day 2 of this case studyit might be wise to reserve one or two for this purpose Circle the tests you would like to order ON DAY 1 Only one order sheet per TEAM Acute Hepatitis Panel lgM antiHAV lgM antiHBc HBsAg HCV RNA Serum AntiDNase B Antibody Serum AntiEBNA Antibody lgG Serum AntiViral Capsid Antigen VCA lgM Serum Antinuclear Antibody IFA FANA on Hep2 substrate Serum Complement C3 Serum Complete Blood Cell Count with Differential Whole blood Erythrocyte Sedimentation Rate Whole blood ESR Human Chorionic Gonadotropin hCG Quantitative Pregnancy Serum Infectious Mononucleosis Rapid Test Serum Lyme C6 Peptide Enzyme immunoassay EIA lgMlgG Serum Lyme Disease Antibody lmmunoblot Serum separate lgM and lgG Rapid Plasma Reagin RPR Serum Rheumatoid Factor RF Serum Latex agglutination immunoturbidometric Syphilis Antibody lgG Serum Treponema pallidum antigen multiplex flow immunoassay Treponema pallidum Particle Agglutination TPPA Serum Justify each test you order write your justification on the back of this page 1 Interpret the test results your team received Do you require any confirmatory tests Please ask for the results of those tests now Use the same team order sheet you used on case day 1 VGA 2 After reviewing all of the laboratory results are you able to make a definitive diagnosis What is the most likely diagnosis MONO FBLD 434 Case 3 SS15 Case History Mary Kettering began experiencing After several days of noticeable Mary called her physician s office and made an appointment Mary had always previously been relatively healthy She had been obese in childhood but lost weight in middle age and had become quite fit Other than that She had just recently retired after 30 years working at the Secretary of State office She was looking forward to winters in Florida playing tennis with friends and working in her garden Mary is divorced has three grown sons and four grandchildren She currently lives in a condominium with her cat Picasso Mary was from a large family She has six siblings three male and three female She was the youngest child Her siblings were all active and healthy with no reported joint painproblems The only common ailment in the family is which four of the six siblings have had or currently have on her scalp when she was in her teens and again in her early 40 s Mary has never been hospitalized She delivered her three sons at home with the assistance of a midwife Since retirement Mary has been teaching yoga at a local YMCA When asked about her current symptoms Mary described the n a day by day basis The equally painful on both sides and it is always most severe in the morning It has become difficult for her to get out of bed in the morning and it can take several hours before she feels better and can move more easily Yoga helps with the pain In addition to the joint pain Her feet and hands also get cold quickly turning purple or white in air conditioned places Despite the pain and stiffness These symptoms have now been present for almost two months and the pain is becoming nearly unbearable Complete Blood Cell Count with Differential lndices Test Result Reference Range WBC x103 lpL 81 40 120 RBC x1061L 461 350 555 Hemoglobin gdL 133 120 150 Hematocrit 382 360 450 MCH pg 289 270 330 MCHC gdL 348 310 370 MCV fL 829 80 100 Platelets x103 lpL 173 150 400 Neutrophils 614 490 810 Lymphocytes 243 140 410 Monocytes 108 0 110 Eosinophils 21 0 60 Basophils 06 0 30 Rheumatoid Factor 9 lUmL Reference lt 15 lUmL Cyclic Citrullinated Peptide CCP AB lgG lt16 Units Reference range Negative lt20 Weak Positive 2039 Moderate Positive 4059 Strong Positive gt59 3315 Case 3 Question Set 1 1 Compare and contrast the methodology specificity and sensitivity of the RF and Cyclic Citrullinated Peptide CCP AB lgG assays Which of these laboratory tests performed on this patient is most specific for the diagnosis of Rheumatoid Arthritis Explain your reasoning a Rheumatoid Factor RF i 80 of patients with RA have a positive RF screen test ii Screening tests for RF are agglutination tests with either RBC or latex particles coated with human lgG 1 Only lgM RF will cause agglutination 2 Not specific to RA patients with syphilis Lupus hepatitis tuberculosis infectious mononucleosis and Sjogren s syndrome can have a positive RF iii lgG RF and lgA RF tested by EIA or nephelometry are much more disease specific and present in nearly 100 of patients b AntiCCP antigyclic gitrullinated peptide is new screening test often paired with RF for new diagnosis of RA with 98 specificity to RA i Citrulline is a nonstandard amino acid created by deamination of arginine by the enzyme peptidylarginine deaminase ii This process is posttranslational modification of joint proteins fillagrin vimentin and fibrin and others 2 Interpret the result for the patient s Erythrocyte Sedimentation Rate What does it indicate in this patient Are there alternate tests more modern that could have been performed that would have yielded comparable information for the physician a The Erythrocyte Sedimentation Rate is outside the range i This indicates there is presence of inflammation ii Creactive protein iii Complement components 3 Does this patient meet the criteria for diagnosis of Rheumatoid Arthritis Why or why not a Yes She has Rheumatoid Arthritis i Large and small joints 5 points 1 Hand 2 2 Feet 2 3 Wrists 2 ii Duration of symptoms two months 1 point iii ESR 1 point b Her points put her within the criteria for diagnosis of Rheumatoid Arthritis you need 6 points CCP for targeting amino acids specifically other testpositives and negatives for other autoimmune diseases 8815 Case 3 Question Set 2 1 What are Seronegative Spondyloarthropathies How do they differ from Rheumatoid Arthritis What is arthritis mutilans and which arthritic diseases is it most commonly associated What are spondyloarthropathies Q Spondyloarthropathies are a family of longterm chronic diseases of joints These diseases occur in children juvenile spondyloarthropathies and adults They include ankvlosino Spondvlitis Reiter39s svndrome reactive arthritis psoriatic arthritis and joint problems linked to inflammatorv bowel disease enteropathicarthritis Spondyloarthropathies are sometimes called spondyloarthritis 0 Although all spondyloarthropathies have different symptoms and outcomes they are similar in that all of them 0 Usually involve the attachments between your low back and the pelvis sacroiliac ioint Q Affect areas around the joint where your ligaments and tendons attach to bone enthesitis such as at the m foot or hip It is important to recognize that the spondyloarthropathies are different from rheumatoid arthritis RA in adults and iuvenile idioDathic arthritis JIA in children a Seronegative Spondyloarthropathies symptoms include i Back pain ii Uveitis iii GI symptoms iv Rashes b Difference between 88 and RA i Rheumatoid factor RF is usually negative if you are expressing spondyloarthropathies symptoms ii RF is usually positive if you are have Rheumatoid Arthritis 1 Individuals with Rheumatoid Arthritis RA will have a positive Rheumatoid factor RF the majority of the time about 80 0 Arthritis mutilans An extremely severe form of chronic rheumatoid orDsoriatic arthritis characterized by resorption of bones and the consequent collapse of soft tissue When this affects the hands it can cause a phenomenon sometimes referred to as 39telescoping fingers 2 Given the patient history and laboratory results as a whole this patient technically meets the diagnostic criteria for Rheumatoid Arthritis What does NOT match a usual presentation of Rheumatoid Arthritis a RF is within the reference range which is weird because if you have RA you usually have a positive RF b CCP should be positive not negative 3 What are Biologic Therapies in the context of arthritis Specifically name the most common biologic therapy used in arthritis patients and describe its mechanism of action Xeljanz was a new drug approved by the FDA in November 2012 for the treatment of Rheumatoid Arthritis What is it s mechanism of action and is it a biologic therapy a A biologic drug copies the effects of substances naturally made by your body39s immune system i Biologic agents are genetically engineered drugs meaning that human genes that normally guide the production of these natural human immune proteins ie an antibody to TNF are used in nonhuman cell cultures to produce large amounts of a biologic drug ii These drugs are given to lessen inflammation by interfering with biologic substances that cause or worsen inflammation These new biologic agents can specifically affect some of the abnormalities of the immune system that lead to the joint inflammation and other abnormalities seen in rheumatoid arthritis and so help treat its symptoms b Xeljanz tofacitinib works by blocking certain enzymes in the body that affect immune system function i Cells in our body talk with each other One way of doing this is through special proteins called cytokines Cytokines are made and released by cells Cytokines then bind to receptors on the surface of other cells This results in a message being sent from the cell surface to the nucleus of the cell called signal transduction This signal results in the production of a protein or peptide by the DNA activation of transcription ii One of the important signal transduction pathways in the body uses an enzyme called Janus Kinase JAK and another molecule called Signal Transducer and Activation of Transcription STAT The JAKSTAT pathway is important for sending signals involving bone marrow and immune system activation iii Tofacitinib is a new small molecule that enters a cell and binds to the ATP binding site on the enzyme Janus Kinase JAK By doing this it prevents JAK from activating STAT and sending the important message to the nucleus of the cell iv With rheumatoid arthritis the immune system is not functioning correctly This causes an immune attack on the joints Tofacitinib Xeljanz interferes with the immune system cells ability to talk with each other When cells can t talk they have a hard time coordinating their attack on the joints The end result is improvement in the arthritis c Xeljanz is used to treat moderate to severe rheumatoid arthritis in adults who have tried methotrexate without successful treatment of symptoms i Without having these symptoms you would not want to give a patient this drug BECAUSE this would trigger LESS inflammation SIDE EFFECT Inflammation is an important process within the immune system as it allows the body to effectively respond using the adaptive immune system alongside the innate immune system Without having this otherwise natural response you would be hindering other processes in the body that would be helping you d Xeljanz is sometimes given in combination with methotrexate or other arthritis medicines BLD 434 8815 Case Study 2 Patient History Eleven month old Evan Ferguson was brought to his pediatrician s office with a and Evan s cervical and axillarv lvmch nodes were not palpable even though he was exhibiting rapid and shallow breaths with raes The pediatrician suspected PP Evan was admitted to the hospital for supportive treatment and additional testing Sputum and blood cultures were He was started on intravenous antibiotics and remained hospitalized for 6 days Results of blood tests ordered by the ER physician on admission can be found below In addition his pediatrician has He had tubes inserted in his ears at 8 months of age to facilitate drainage but has Evan has a includina an Optional vaccine for Haemochius influenza Patient Evan Ferguson Age 11 months Sex Male Eemplete Bleed Eell Eeunt lndiee Test Result Hefielrenee we is 153 eellllsiluL 513 55 11 5 es s mt eelllsiluL 411 512 Hemegllsllzlliln gmij 12 52511 1561 MEH its 391 ET MEHE QML 32 mes 11 31 1 61 92 Pietellets s 1 E1 5 15 451 Nevutreshills 1 2245 Nevutrephil sends 1 s 1 3 es Lylmyslheeytes 1 TE 3 Meneeytes 193 E 211 Eeslilnephile 113 1 elseehils 1 El 1 192 Tetel EEll39llJIITI Antilhedies lsetylpe Test esu It HEfEll39EIl IEE es mes Eli 151313 quotgait mgl L 5 5451513 gs lmgldL see 515 i Analysis of lsohemagglutinins Evan is blood type A lgM titer of antiB antibodies was positive at 1160 lgG antiB antibody was undetectable Alleselute Peripheral leed Lympheeyte Lieu nt Test Result Reference 11115 eellelL j sear T sells 11 nil12 est T GENE 1i 4 21 El lly39lmplheeytes 1J1 ICIIL1EEJlr Tetell llylmplheeytes 4113 1n412u1 lgG mgdL 45 RR 6001500 lgA mgdL lt5 RR 50150 lgM mgdL 230 RR 75150 Day 1 Discussion Questions 1 What immune cells are directly and indirectly involved in the production of the three total serum antibodies tested for in Evan s serum In listing these explain the general role of each of these cells Q Helper Tcells CD4 Tcells Q MHCll Q lgM Q lgA Q lgG 2 Considering your answer to question 1 which of these immune cell types could have a defect which could explain the findings observed with Evan Explain your reasoning for thisthese choices a It seems like there is a problem with the interact interaction that inhibits isotype switching see below b We say this because lgM can still be produced which means that MHCll is working fine enough for the Bcell to interact successfully with the TCR i However isotype switching is flawed that makes lgG and lgA almost nonexistent lgG is higher in trace amounts because of having had lgG transferred passively through the placenta For this reason it is best to assume that CD40 and CD4OL are not working properly Day 2 Discussion Questions 1 Evan was diagnosed with infection with Streptococcus pneumoniae Describe the characteristics of this infectious agent and the immune effector cells andor proteins that would likely be involved in ridding the body of this organism Are you surprised this patient has neutropenia Q It is also the most common cause of communityacquired pneumonia and pinkeye l Anyone can become infected l However people who are at for the infection are 0 The elderly 0 Children who attend group day care centers and people with underlying medical conditions l Symptoms depend on type of infection 0 If develops symptoms may include ear pain fever balance problems trouble sleeping and change in behavior 0 lf develops symptoms may include sudden onset of high fever productive cough headache and shortness of breath O lfdevelops symptoms may include discharge that may cause the lids to stick together especially after sleeping redness tearing and a gritty feeling l Symptoms may appear in one to three days However children are more likely to carry this bacterium around in their noses and throats and do not have any symptoms I Q There is no persontoperson spread however before and after cleaning the ear or adding drops it is always good practice to wash hands thoroughly I 0 People don t spread pneumonia mostly through respiratory droplets from their nose or mouth even if they don t have any symptoms 0 People can get pinkeye by coming into contact with tears or discharges from the eyes of an infected person and then touching their own eyes I This will depend on the illness With pneumonia a person is usually no longer contagious after 24 to 48 hours of antibiotics For other illnesses consult with a healthcare provider for prevention guidance l Diagnosis depends on the illness l Most people with Streptococcus pneumoniae are treated with antibiotics Q A person does build immunity to S pneumoniae however there are many different types of S pneumoniae and one type does not cause immunity for another So in a person s lifetime he or she may become infected with several different types of S pneumoniae l Children should be excluded from a childcare setting if the illness prevents the child from participating or if the child has a fever lethargy irritability persistent crying or shortness of breath If the staff feels that they are jeopardizing the health andor safety of other children in the group by having to care for the child the child also may be excluded l Children with pinkeye that has white or yellow discharge eye pain and redness of the eye and surrounding tissue should be excluded until examined by a physician and approved for readmission P l A vaccine is available for the prevention of pneumococcal disease in infants and toddlers The vaccine PCV 13 Prevnar protects against thirteen different types of S pneumoniae Children are recommended to receive four doses at 2 4 6 and 12 to 15 months of age The vaccine has been shown to be highly effective in preventing ear infections and pneumonia l A vaccine also is available for adults This vaccine PPV23 Pneumovax protects against 23 different types of S pneumonia All adults age 65 and older should receive one dose Also anyone age 2 to 64 who is at high risk for pneumococcal disease should receive a dose Contact your health care provider to see if you are at high risk O Creactive protein also binds to chammaRl and FCgammaRll and by this mechanism it delivers Streptococcus pneumoniae for uptake and degradation by phagocytes O CD4OL O Neutrophils Neutrophils or polymorphonuclear leukocytes comprise a crucial component of innate immunity controlling bacterial and fungal infection through a combination of both oxidative and nonoxidative mechanisms Neutrophils are believed to play an important role in caused by the major human pathogen Streptococcus pneumoniae O GMCSF 0 White blood cells can also be a target for autoantibodies and complement activation Because nucleated cells S pneumoniae are less susceptible than erythrocytes to complementmediated lysis the major effect of complement activation is to facilitate the phagocytemediated clearance of white blood cells in the spleen 0 Expression of a surface endonuclease encoded by endA 21 is a common feature of many pneumococcal strains We show that EndA allows pneumococci to degrade the DNA scaffold of NETs neutrophil extracellular traps and escape Furthermore we demonstrate that escaping NETs promotes spreading of pneumococci from the upper airways to the lungs and from the lungs into the bloodstream during pneumonia 2 Keeping in mind your answer to question 1 is there any evidence that Evan has a defect in any of these cells andor proteins secreted or surface ligandsreceptors Specifically can you rule out any of them as a possible cause Why or why not 0 Because the scaffold of NETs is DNA we tested whether pneumococcal EndA affects NET integrity Activated neutrophils were incubated with either culture medium RPMI bovine pancreatic DNase TlGR4AendA or TIGR4 After 30 min incubation the samples were fixed and stained for DNA blue and NE red NETs were intact in cells incubated with medium In contrast infection with TlGR4AendA did not affect the NET integrity 0 GMCSF 0 Since there are no neutrophils being produced it can be inferred that there is a problem with GMCSF because the cytokine produces neutrophils I That also means that there is a problem with the CD8 Tcells because the interaction between the CD40L and CD40 on the other Tcells were not working properly which would mean that the CD40L and CD40 receptors would not work properly on these cells either 0 LPS 0 LPS is a Tl1 antigen that activates LPS specific Bcells I This makes lgM in excess which is why Evan has a lot of lgM 0 Pages 109 116 117 3 Is it possible that Evan could have AIDS Acquired Immune Deficiency Syndrome due to HIV infection or SCID Be sure to explain your reasoning for your answer Topic 1 Immune Cells Innate and Adaptive Immunity Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to Q Antigen A toxin or other foreign substance that induces an immune response in the body especially in the production of antibodies 0 Innate immunitv Immunity possessed by a group as a species or race that is present in an individual at birth prior to exposure to a pathogen or antigen and that includes components as intact skin salivary enzymes neutrophils natural killer cells and complement that provide an initial response against infection 0 Response is RAPID hours 0 Fixed 0 Limited number of specificities O Constant during response 0 Adaptive immunitv Immunity usually found in higher life forms like eukaryotic organisms that protects against infectious disease agents through B and T lymphocytes following exposure to a specific antigen 0 Response is SLOW days to weeks 0 Variable 0 Numerous highly selective specificities 0 Improve during response 0 CD antigen marker A system commonly used in immunophenotyping allowing cells to be defined based on what molecules are present on their surface These markers are often used to associate cells with certain immune functions 0 Sebum An oily secretion of sebaceous glands Q Defensins Small cysteine rich cationic proteins found in both vertebrates and invertebrates Also found in some plants They function as host defense peptides and are active against bacteria fungi and many enveloped and nonenveloped viruses Q Lysozyme An enzyme that catalyzes the destruction of the cell walls of certain bacteria occurring notably in tears and egg white 0 Polvmorphonuclear Having a nucleus with several lobes and a cytoplasm that contains granules ie eosinophil basophil neutrophil 2 Characterize Innate and Adaptive immunity in terms of the speed of response the amount of variability of antigen recognition the level of specificity of antigen recognition the ability to improve during a response andor with repeated exposure INNATE IMMUNITY ADAPTIVE IMMUNITY Response RAPID hours Response SLOW days weeks Fixed antigen recognition Variable antigen recognition Limited number of specificity Numerous highly selective specificity Constant during response Improve during response 3 List the common CD markers that can be used to identify T lymphocytes including the 2 main subsets and B lymphocytes 0 B Lymphocytes Plasma cells CD19 and CD20 Q T Lymphocytes CD3 0 Th CD4 O TC CD8 O Treg CD4 4 List the mature immune cell types found in blood and tissues that can arise from the common lymphoid progenitor and the common myeloid progenitor Q The common lymphoid progenitor can produce BCells TCells and NK cells 0 The common myeloid progenitor can produce neutrophils basophils eosinophils monocytes dendritic cells macrophages and mast cells 5 Explain the relationship between monocytes and macrophages 0 Monocytes are found in blood circulation and macrophages are found in tissues but are essentially both the same type of cell 6 List the 3 polymorphonuclear leukocytes granulocytes found in peripheral blood and the color of cytoplasmic granules each possesses 1 Eosinophils gt OrangePink 2 Basophils gt DarkPurpIeBlack 3 Neutrophils gt PinkishBlue Topic 2 Adaptive Immunity Lymphoid Tissues Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to Q Primarv lvmphoid tissue bone marrow and thymus Where lymphocytes develop antigen receptors and undergo testing to make sure they are going to be functional and not harmful to self when they are released into the tissue Basically where T and B cells are made 0 Secondarv lvmphoid tissue where lymphocytes are activated by the antigen Primarily spleen and lymph nodes 0 Lymph a clear yellowish slightly alkaline coagulable fluid containing white blood cells in a liquid resembling blood plasma that is derived from the tissues of the body and conveyed to the bloodstream by thelymphatic vessels 0 Nai39ve lvmphocvte immune cells that are mature but have not yet been exposed to anantigen These cells move freely through the immune svstem and play an important role in the development and maintenance of immunity They arrive at the lymph nodes via arterial circulation arterioles and exit via lymph Q PALS Deriarteriolar lvmphoid sheath A portion of the white pulp of the spleen They are populated largely by T cells and surround central arteries within the spleen PALS Tcells are presented with blood borne antigens via myeloid dendritic cells 0 GALTgut tonsils adenoids appendix and peyer s patches 0 BALT Respiratory tract immune system AN example of mucosa associated lymphoid tissue 0 MALT Mucosal immune system The diffusion system of small concentrations of lymphoid tissue found in various sites of the body like the GI tract thyroid breast lung salivary glands eye and skin 0 M cell microfold deliver pathogens across epithelium 2 List the 2 tissuesorgans that represent the primary lymphoid tissues 0 Bone marrow and thymus where lymphocytes are made 3 Describe how a nai39ve lymphocyte enters a lymph node and how it would exit if it did not get stimulated by antigen 0 Naive lymphocytes arrive at lymph nodes via arterial circulation not through lymph fluid 0 Enter via arterioles blood vessels exit via efferent lymph 4 Describe how antigen enters a lymph node 0 Antigens enter lymph nodes via fluid draining from tissues in lymph vessels carried by dendritic cells that crawl into the lymphatic vessels to the lymph node 5 List where T lymphocytes and B lymphocytes are primarily located in the lymph nodes and the white pulp of the spleen Q T Cells located primarily in PALS surrounding a central arteriole Q B Cells organized in germinal center and B cell corona peripheral to PALS 6 What are the main advantages of the adaptive immune response Q It has immune memory so its response improves It also has numerous highly selective specificities 7 Describe the primary and secondary antibody responses to an antigeninfection including the relative amount of antibody produced the speed at which the antibody is produced hours or days and in the process define what is meant by lag phase 0 Primary weak less antibody lag phase 0 Secondary immediate more antibody no lag phase 0 Lag Phase time where B Cells proliferate and differentiate where no antibodies are produced Primary Response Secondary Response Antigen Slow Rapid Antibody Level Low High Rate of Antibody Slow Rapid Production Lag Phase Yes No pg 4 Topic 3 Innate Immune Sensing of Danger Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what ces the term applies to as appropriate phagocyte C opsonin anaphylatoxin chemoattractant complement fixation and CR 0 Phagocyte cell specialized to perform phagocytosis The principle phagocytic cells in mammals are neutrophils and macrophages Cells that specialize in the capture engulfment and killing of microorganisms Q Q Complement The first line of defense against an extracellular infection A large family of soluble proteins produced primarily by the liver almost all are zymogens inactive enzymes many of which are serine proteases because they are enzymes complement products activate each other in a cascade fashion with tremendous amplification capability Q Opsonin Antibodies and complement components that bind to pathogens and facilitate their phagocytosis by neutrophils or macrophages 0 Anaphvlatoxin Complement fragments C3a and C5a which are produced during complement activation They act to induce inflammation recruiting fluid and inflammatory cells to sites of antigen deposition In some circumstances C3a and C5a can induce anaphylactic shock Q Chemoattractant Complement fragments can recruit immune cells to sites of inflammation by creating a concentration gradient that the cells follow 0 Complement fixation The covalent attachment of C3b or C4b to pathogen surfaces which is a central feature of the action of complement because it facilitates phagocytosis of the pathogen 0 CR complement receptors Bind iC3b and various other surface molecules of bacteria and yeast 2 Explain what is meant by the following statement Complement works in a cascade fashion that amplifies as it proceeds 0 Infection triggers complement activation which proceeds by a series or cascade of enzymatic reactions involving proteases in which each protease cleaves and activates the next enzyme in the pathway 3 List the major advantage that complement has compared to antibody in tagging pathogens for disposal Q Complement fragments can covalently bind to pathogen surface so a permanent attachment whereas antibodies only attach using a noncovalent bond which is reversible 4 List and define the 4 main functions that the complement proteins can perform for the immune system a Opsonization when complement fragments can covalently bind to pathogen surface or soluble immune complexes tagging them for uptake by complement receptors b Anaphylatoxin complement fragments can diffuse away and act as inflammatory mediators Toxic products that can induce anaphylaxis in high concentrations c Chemoattractant complement fragments can recruit immune cells to sites of inflammation by creating a gradient that the cells follow d Lysis completion of complement cascade can create a pore in the membrane to bacterial lysis MAC 5 Name the 3 separate pathways that can trigger complement activation and identify the first complement components that binds to initiate each of these pathways 1 Alternative pathway formation and action of the soluble C3 convertase iC3Bb that initiates it Spontaneous C3 hydrolysis 2 Lectin pathway mannosebinding lectin binds to pathogen surface 3 Classical pathway Creactive protein or antibody binds to specific antigen on pathogen surface 6 Identify the actual complement proteins that are complexed together to form the C3 convertase and C5 convertase for both the classical and alternative complement pathways 0 C3 Convertase In the plasma close to a microbial surface the thioester bond of C3 spontaneously hydrolyzes at low frequency This activates the pathway causing C3 to bind to factor B Cleavage of B by the serine protease factor D produces a soluble C3 convertase called iC3Bb which then activates C3 molecules by cleavage into C3b and C3a Some of the C3b fragments become covalently attached to the microbial surface 0 C5 convertase The CS convertase of the alternative pathway consists of two molecules of C3b and one of Bb C3BZBb CS binds to the C3b component of the convertase and is cleaved into fragments C5a and C5b of which C5b finitiates the assembly of the terminal complement components to form the membraneattack complex 7 Explain what the similarities and differences are between iC3 versus iC3b as well as iC3Bb versus C3bBb being certain to identify which are enzymatically active and which have been inactivated a iC3 and iC3b both inactive iC3b contains factor b which is an opsonin easier to phagocytize Both part of alternative pathway b iC3Bb and C3bBb both active both C3 convertases difference is the iC3Bb is soluble whereas the other fixed and in the tissue 7 i Bb think ACTIVE 8 Identify which complement component fragment provides the greatest amount of amplification of the complement cascade and why ie C2b C3b C4b C5b Q C3b because it binds factor B to bind more factors and amplify the cascade 9 Identify the complement components that compose the membrane attack complex MAC or also known as terminal complement components O C5C9 0 C9 is the one that forms the actual pore 10 Identify which complement component actually polymerizes to form a pore in the pathogen membrane C9 11 Identify the single protein that can amplify complement activation what pathway of complement it amplifies and the mechanism it uses to accomplish amplification pg 5 Q Proloerdin factor P is a plasma protein that binds to C3bBb alternative C3 convertase bound to pathogen surface and prevents its degradation by proteases This keeps the alternative pathway active longer 12 List the two points at which complement activation can be inhibited and identify the product of C3b cleavage produced by Factor I 1 lnactivate C3 convertase a Soluble Factor H binds to C3b targeting it for cleavage by Factor 1 b Membrane Decayaccelerating factor and membrane co factorMCP bind to C3b of C3bBb and dissociates the complex But MCP targets C3b and makes it susceptible by cleavage from factor 1 2 Prevent MAC formation a 8 protein clusterin and factor J prevents complex association with cell membrane b Homologous restriction factor and CD59 prevent C9 recruitment iC3b is the product 13 Describe how a2macroglobulins and defensins function in innate immune system protection of the body 0 Alpha 2macroglobulins qucoproteins with molecular mass 180 kDa and act as protease inhibitors 0 Degraded protease bound to this is picked up by receptors present on hepatocytes fibroblasts and macrophages 0 Defensins a family of small amphipathic proteins that disrupt microbial membranes O Produced primarily by Paneth cells base of crypts epithelial cells and phagocytes 14 List the various receptors phagocytes possess that allow them to recognize and phagocytize pathogens 0 Lectins O Mannose receptor 0 Glucan receptor 0 Scavenger receptor binds negatively charged ligands such as sulfated sugars nucleic acids and phosphate containing lipids of gram bacteria 0 Complement receptors CR3 CR4 bind iC3b and other surface molecules of bacteria and yeast 15 Identify the role of TLR in innate immune responses Tolllike receptors recognize an array of microbial components 0 They are differentially located in cell or endosome membranes 0 TLR signaling activates cytokine gene expression in infected cell 0 Viral infection gtTLR3 gt type 1 lFNs only alpha beta 0 Bacterial infection gt multiple TLRs gt inflammatory cytokines TNF alpha lL1 beta and lL6 pg 6 Topic 4 Innate Responses Phagocytes and NK Cells Induce Local Acute Inflammation Objectives At the completion of the modules and the assigned reading the student should be able to 1 Describe how neutrophils and macrophages differ in their lifespan and function as phagocytes Q Macrophages 0 Resident in all tissues and are usually the first immune cell to respond to pathogen entry 0 Are phagocytic but longlived and clean up many dying cells and tissue debris 0 They also secrete cytokines that influence adaptive immunity 0 Neutrophils 0 Enter inflammatory sites in large numbers phagocytize and digest pathogens bacteria and then die pus 2 Identify the cytokines released by macrophages that cause beneficial LOCAL immune system effects and the specific effects that each of the cytokines is responsible for Q TNFor O Activates vascular endothelium and increases vascular permeability which leads to l Increased entry of complement and cells to ssues l Increased fluid drainage to lymph nodes 0 lL1B O Activates vascular endothelium O Activates lymphocytes 0 Local tissue destruction 0 Increases access of effector cells 0 CXCL8 O Chemotactic factor recruits neutrophils and basophils to site of infection 0 lL12 O Activates NK cells is produced in greater amounts during a viral infection 0 IL 6 has no local effect All of its effects are systemic 3 Describe how acute systemic activation of macrophage cytokine production can lead to septic shock Q Macrophages can secrete too much of a good thing 0 Systemic macrophage activation such as in bacteremiasepticemia can lead to septic shock and death All of the macrophages in your body being activated at once 0 Ruptured appendix 0 Very invasive bacterial infection 0 Septic shock primarily mediated by excess TNFOt O Macrophages activated in the liver and spleen secrete TNF into the bloodstream 0 Then systemic edema causes decreased blood volume hypoproteinemia and neutropenia followed by neutrophilia Decreased blood volume causes collapse of vessels 0 Disseminated intravascular coagulation leads to wasting and multiple organ failure septic shock 4 Define chemokines and the two signature amino acid sequence motifs that characterize the chemokine family 0 Chemokines a large family of small proteins secreted by immune cells and many nonimmune cells stromal cells keratinocytes endothelium fibroblasts O Secreted in sites of inflammation 0 Attract immune cells 0 Chemokines are characterized by a sequence motif containing adjacent cysteines O CXC chemokines ex CXCL8 produced by activated macrophages attract neutrophils and naive T cells and can control angiogenesis 0 CC chemokines attract monocytes effector T cells and memory T cells I CXC and CC 5 Describe in general what a cell that expresses a chemokine receptor does in response to binding it s chemokine ligand 0 The process of extravasation is promoted by chemokines and adhesion molecules 0 Cells roll on endothelium using addressin binding to selectins 0 Tight binding of cell to endothelium is caused by integrin binding to immunoglobulinIike molecule 0 Cells crawl up chemokine concentration gradient area of lesser concentration to area of greater concentration 6 Explain what is meant by respiratory burst in regards to phagocytes Q Granules fuse to phagosome Q NADPH oxidase enzymes consumes molecular oxygen in driving an electron transport train which removes H in phagosome raising pH 0 Increased pH releases defensins and activates neutral proteases concentrated in granules 7 Identify the immune system defect that causes Chronic Granulomatous Disease 0 Chronic granulomatous disease results from defects in NADPH oxidase subuan 0 Respiratory burst is defective and granule enzymes are not activated 0 Infectious bacteria and fungi are phagocytosed by neutrophils and macrophages but survive inside the phagosomes Q Infected cells form a localized reaction called granuloma to wall off infection 8 Identify the major macrophage cytokine that triggers the acute phase response and the proteins that are produced as a part of the acute phase response 0 Triggered by lL6 0 Proteins produced 0 Mannosebinding Iectin MBL O Creactive protein CRP Q Fibrin NEEDS TO BE REPLACED O Clotting O Trapping bacteria 9 Describe the importance of an acute phase reaction response and which tissuesorgans of the body are primarily responsible go beyond the liver here Q The acutephase response increases the supply of the recognition molecules of innate immunity 0 Acutephase proteins are produced by liver cells in response to the cytokines released by phagocytes in the presence of bacteria In humans they include Creactive protein fibrinogen and mannosebinding lectin Both C reactive protein and mannosebinding lectin bind to structural features of bacterial cell surfaces that are not found on human cells On binding to bacteria they act as opsonins and also activate complement facilitating phagocytosis and also direct lysis of the bacteria by the terminal complement components 10 Identify which of the interferons lFNu lFNB or lFNv are Type I interferons and the type of microbial infection they combat ie Bacterial Viral Fungal Parasitic Q lFNaB are Type I lnterferons 0 They inhibit viral replication by proteases that degrade viral RNA 0 They increase antigen presentation to activate cytotoxic T lymphocytes of adaptive immunity 0 They activate NK cells to kill virusinfected cells 11 Explain how NK cells determine whether a host cell is dangerous and must be killed 0 The balance of inhibitory and activating signals received from a cell determines whether an NK cell will target it for killing 0 NK cells express diverse receptors from two families 0 lmmunoglobulinlike receptors 0 Lectinlike receptors 12 Identity how activation of the classical lectin and alternative pathways of complement can each be initiated specifically listing the factors that do such What step of complement activation do these pathways all converge and why is this important 0 Classical 0 CRP or AB C1qrs C4 C2 0 Lectin O MBL C4 02 i I all the of mm plge ifizati cfa n hem and Ana atoxin and 10 mars pg 7 Topic 5 Overview of Adaptive Immunity Learning Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to epitope antigenic determinant peptide somatic recombination positive selection negative selection clonal selection clonal expansion antibody Ab immunoglobulin lg antibody class antibody isotype humoral immunity cellular cellmediated immunity BCR TCR and MHC major histocompatibility complex 0 Epitope O The portion of an antigenic molecule that is bound by an antibody or gives rise to the MHCbinding peptide that is recognized by a Tcell receptor Also called an antigenic determinant Q Antidenic determinant T and B cell receptors are specific for a single epitope or portion of an entire antigen 0 Peptide 0 Proteins that have been broken down Usually 925 AA long 0 Somatic Recombination 0 DNA recombination that occurs between gene segments in the immunoglobulin genes and Tcell receptor genes in developing B cells and T cells respectively It generates a complete exon composed of a V gene and a J gene segment and a D gene segment at the immunoglobulin and Tcell receptor heavychain loci that encodes the variable region of an immunoglobulin or Tcell receptor polypeptide chain 0 to make variable antigen binding receptors 0 Positive selection 0 Process occurring in the thymus during Tcell development that selects immature T cells with receptors recognizing peptide antigens presented by self MHC molecules Only cells that are positively selected are allowed to continue their maturation 0 Negative selection 0 Process in the thymus whereby developing T cells that recognize self antigens are induced to die by apoptosis 0 Clonal selection 0 The central principle of adaptive immunity It is the mechanism by which adaptive immune responses derive only from individual antigenspecific lymphocytes which are stimulated by the antigen to proliferate and differentiate into antigenspecific effector cells 0 Clonal expansion 0 In primary immune response antigenspecific B and T cells proliferate Q Antibodv Ab 0 Molecule that a specific B or T cell receptor recognizes and binds to noncovalent Same as an immunoglobulin Q lmmunoolobulin Id 0 Same as an antibody 0 Antibodv class Q Isotope Five classeslgG IgM IgD IgE and IgA 0 Antibodv isotvpe 0 Class Five classeslgG IgM IgD IgE and IgA 0 Humoral immunitv O Immunity due to antibodies 0 Cellular cellmediated immunitv O Immune response that doesn t involve antibodies but rather involves the activation of phagocytes antigenspecific cytotoxic T and the release of various cytokines in response to the antigen 0 0 B cell receptors B cell receptors and secreted antibodies are not created equal 0 TCR Q Do not have five classes they generate functional diversity by the proteins they associate with TCR s associate with CD4 or CD8 protein on the T cell surface CD8associated TCR s bind to MHC helper and regulatory T cells 0 MHC maior histocompatibilitv complex 0 Antigen presenting molecule Presents peptide fragments to T cells 2 Describe the type of antigens that T lymphocytes and B lymphocytes are able to bind Q T cell receptors recognize peptide fragments Q B cell receptor recognize primary secondary tertiary or quaternary structures of proteins glycoproteins Iipoproteins or proteoglycans 3 Be able to draw and correctly label the basic B lymphocyte receptor and T lymphocyte receptor structures identifying the light chains heavy chains variable regions constant regions or chain 3 chain and the antigenbinding sites T Cell and B Cell Antigen Receptors TCR and BCR B cell Receptor T cell Receptor BClR Tc recognition g m recognition 7 i li chain A heavy clhain 3993 In signalling signaling will need to know for exam 4 Identify whether the variable regions or constant regions of T cell and B cell receptors function in antigen binding or are responsible for the effector function The variable regions of T and B cell receptors function in antigen binding Specialized function of Ig molecules varies based on constant region Constant region determines antibody class 5 List the approximate number of T lymphocytes or B lymphocytes that should be able to respond to a given antigen the first time your body is exposed to that an gen Lymphocyte recirculation to up our odds to have lymphocyte that is activated and that will recognize the antigen Few cells per million rare 6 List and briefly describe the 2 main functions of antibodies Neutralization inhibit cell entry coat toxins Opsonization coat for uptake by phagocytes 7 List the two T cell coreceptor molecules and whether they bind to MHC Class I or MHC Class II molecules CD8 TCR s bind to MHC l CD4 TCR s bind to MHC II 8 List the main function for each of the 3 effector T cells Th1 Th2 Tc and be able to identify whether they possess CD4 or CD8 on their surface T cells can aid in both cellular Th1 and humoral immunity Th2 as well a accomplish cellular immunity Tc Tc has CD8 on its surface Th1 amp Th2 have CD4 on their surface pg 8 Topic 6 B Cell Receptor Immunoglobulin Structure pg 95112 Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to CDR hypervariable region RSS TdT RAG enzyme combinatorial diversity junctional diversity P nucleotides and N nucleotides monoclonal antibody antisera O CDR O Acronym for complementaritydetermining regions the antigen binding loops of immunoglobulins antibodies and Tcell receptors HVIoervariable region 0 Small regions of high amino acid sequence diversity within the variable regions of immunoglobulin and Tcell receptors They correspond to the complementaritydetermining regions R88 O Acronym for the recombination signal sequences flanking the V D and J segments in immunoglobulin and Tcell receptor genes 0 RSSheptamerspacer12 or 23 spacernonamer m 0 Acronym for terminal deoxynucleotidyltransferase Enzyme that inserts nontemplated nucleotides N nucleotides into the junctions between gene segments during the rearrangement of the heavy chain genes for Tcell receptor and immunoglobulin RAG enzvme O Recombination Signal Sequences RSS s direct the RAG enzymes that cut the DNA between the gene segments 0 RAG enzymes bind and cut at RSS s Combinatorial diversitv 0 created by somatic recombination of randomlyselected gene segments Junctional diversitv 0 Diversity present in immunoglobulin and Tcell receptor polypeptides that is created during the process of gene rearrangement by the addition or removal of nucleotides at the junctions between gene segments 0 Occurs at the cut sites joining the VDJ segments O Adds a factor of about 3 x 107 to overall lg diversity 0 P nucleotides and N nucleotides 0 P nucleotides 0 Nucleotides added into the junctions between gene segments during the somatic recombination that generates a rearranged variableregion sequence They are an inverse repeat a palindrome of the nucleotide sequence at the end of the adjacent gene segment which gives them their name of palindromic or P nucleotides 0 N nucleotides 0 Nucleotides added at the junctions between gene segments of Tcell receptor and immunoglobulin heavychain variableregion sequences during somatic recombination which contribute to the diversity of these molecules They are not encoded in the gene segments but are inserted by the enzyme terminal deoxynucleotidyltransferase TdT Q Monoclonal antibodv 0 Antibodies produced by a single clone of B lymphocytes and which are therefore all identical in structure and antigen specificity Q Antisera O Fluid component of clotted blood from an immune individual that contains antibodies against a given antigen Antiserum contains a heterogenous collection of antibodies that bind the antigen 2 List the only effector function of B lymphocytes 0 The only effector function of B lymphocytes is antibody production 3 For an antibody molecule tell how many heavy and light chains compose it how these chains are held together where the hinge region is and what the function of the hinge region is 0 Two identical heavy chains and two identical light chains 0 Purpose of the hinge region flexible amino acids Can move the antigen binding domains in relation to each other 0 Hinge region is in the constant region Q There are disulfide bonds that hold the chains together 4 Describe the basic protein folding structure that defines a lglike domain and why it is commonly found in immune system proteins 0 lglike domains are very stable against harsh conditions pH salt proteases Q 3 exposed loops at the end of the V lglike domain form the Agbinding site hypervariable regions or CDRs Q Antiparallel Bsheet lglike domains 5 List the main gene segments that must be spliced together to form either a antibody light chain or a antibody heavy chain 0 Light Chain Composed of a variable a J and a constant gene segment 0 LVJC Q Heavv Chain Composed of a variable a J a constant and a D gene segment D gene segment Add diversity to the gene sequence 0 LVDJC 6 List the two types of light chains that may be used in the formation of an antibody molecule note they are in separate gene loci on separate chromosomes 0 Lambda 0 kappa 7 Describe the relationship between VL VH CL and CH protein domains and the general function of each 0 Variable light Vl VJ form the antigen binding CDR antigen binding site 0 Variable heavyVh VDJ form the antigen binding CDR antigen binding site 0 Constant light aren t antigen binding sites know locations no Constant heavy aren t antigen binding sites know locations study pic 0 The heavy chains determine the antibody class Q The light and heavy chains both contribute to Ag binding sites 8 Describe the significance of heptamers nonamers and 12 and 23 basepair spacers in the targeting of gene segments used to create functional BCR and TCR and the utility of the 1223 rule 0 RSS s flank every V D and J gene segment 0 RSS s are identical heptamers and nonamers with either 12 or 23 nucleotide spacers 0 RSS heptamer spacer nonamer Q Heptamers 7 nucleotides 0 Nonamers 9 nucleotides 0 A 12 spacer can only join to a 23 spacer O This 1223 rule ensures the properjoining order of gene segments so a VH can notjoin directly to a JH skipping a DH segment in heavy chain rearrangement 9 Identify which 2 C gene segments are closest to the variable gene segments in the Ig heavy chain locus and the significance of this location 0 For the heavy chain the C region closest to VDJ contains both CU and C6 0 Naive B cells transcribe through both CU and C6 10 Identify which lg molecules are expressed on the surface of nai39ve B lymphocytes 0 Alternative splicing of mRNA allows naive B cell to express both lgM and lgD BCR simultaneously on the cell surface all of the same antigenic specificity on one cell 11 List the proteins that compose a complete BCR including the signaling molecules 0 A full length BCR is formed by joining a rearranged VDJ to a C region Q C region contains Cu and 06 Q BCR associates with Igor and lgB signaling molecules pg 9 Topic 7 Improving Antibody Affinity and Function Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to Affinity affinity maturation clonal selection and somatic hypermutation Q Affinity measure of the strength with which one molecule binds to another at a single binding site 0 Affinity maturation the increase in affinity of the antigenbinding sites of antibodies for antigen that occurs during the course of an adaptive immune response occurs as a result of somatic hypermutation of the rearranged immunoglobulin region and the consequent selection of mutated B cells that make antigen receptors of higher affinity for their antigen 0 Clonal selection the central principle of adaptive immunity the mechanism by which adaptive immune responses derive only from individual antigenspecific lymphocytes which are stimulated by the antigen to proliferate and differentiate into antigenspecific effector cells 0 Somatic hypermutation mutation that occurs at high frequency in the rearranged variable region of DNA of immunoglobulin genes in activated B cells resulting in the production of variant antibodies some of which have a high affinity for the antigen 2 Identify which of the following processes uses alternative RNA processing to occun Uses Alternative RNA If Yes Which Processes Processing Somatic Recombination No DNA Somatic Hypermutation No DNA Junctional Diversity No DNA lgM and lgD Surface Yes RNA splicing Expression on Naive B Cell lsotype Switching No DNA Production of Membrane Yes Bound BCR Vs Secreted Antibody DNA is permanent and RNA is changeable 3 List what factors primarily control which isotype of antibody will be produced by a B lymphocyte Isotope switchingregulated by cytokines primarily from CD4 T cells 4 Describe the process of isotype switching and whether affinity of antigen binding is improved by this process DNA Rearrangement that matches the rearranged V region with a new C region Antigen binding of the antibody does not change function of the antibody molecule body changes because of different C region different isotype 0 Process 0 1 ND induces cytidine to uracil in switch sequences 0 2 DNA repair enzyme UNG removes base from uracil nucleotide O 3 Endonuclease APE1 excises the abasic nucleotide leaving a nick O 4 Nicked switch sequences associate to promote DNA strand recombination 5 Explain why lgM must be a pentamer to be effective Bc by itself lgM has low affinity so the increased number of potential binding sites helps increase affinity of lgM 6 List the 5 main isotypes classes of antibody and identify which antibody isotypes hashave the following features lgM lgD lgG lgA lgE httoduizletcom5333923bld434test1flashcards lgM lgD lgG lgA lgE Biggest Molecular x Weight Most Abundant Serum amp Body x Fluids Exists as a Pentamer The 3 that Compose 99 x of All Antibody x two x in BloodBody subtypes Fluids Can Cross Placenta Highest Concentration x in Mucus 2 that can Bind to Complement 7 Identify which antibody isotypes and subtypes as appropriate are best at performing each of the following antibody functions lgM lgD lgG lgA lgE Neutralization x x Opsonization x Sensitive for X NK Killing Sensitive for x mast cells Activate x x Complement 8 Identify which antibody isotype can be produced as either a monomer or dimer and which tissue fluids each would be found in Monomer lgA diffusion into extravascular sites Dimer lgA transport across epithelium pg11 Topic 8 T Cell Receptor Structure Objectives At the completion of the modules and the assigned reading the student should be able to 1 Correctly identify whether the following processes occur in T lymphocytes B lymphocytes neither or both T lymphocytes B lymphocytes Somatic Recombination Yes Yes Junctional Diversity Yes Yes Somatic Hypermutation No Yes lsotype Switching No Yes Affinity Maturation No Yes 2 Identify which of the V D and J gene segments are used to create the Vd VB VH and VL protein domains eg Va is composed of only V and J gene segments Composition V D J Vd X X VB X X X VH X X X VL X X 3 Correctly identify Vd VB VH VL Cd CB CH and CL as belonging to the BCR or TCR TCR BCR Vor X VB X VH X VL X Cor X CB X CH X CL X 4 List the proteins that compose a complete functional TCR including the signaling molecules TCRalphaL V J C and beta L V D J C antigen recognition CD3 signaling complex 2 zeta chains 2 CD3 epsilon 1 CD3 delta 1 CD3 gamma CD4 if a T helper cell or T regulatory cell or CD8 if a T cytotoxic cell TCROLB complex with CD3 molecules to transduce signal CD cm CD3 signaling complex 2 TT TT c035 dlelta CDBy gamma 9 9g Del 9 2CD33 epsilon and I I I 2 C zeta chains signalguction 5 Describe unique features and functions of V6 T cells whether these cells express CD3 CD4 or CD8 and how they recognize antigen They are associated with CD4 or CD8 and a CD3 signaling complex Whether they are associated with CD4 or CD8 depends if they were presented with an antigen from the MHC1 or MHC2 complex TCR only recognize antigen as peptide fragments pg 12 Topic 9 MHC and Antigen Processing Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to if appropriate 0 endogenous produced inside the cell in the cytosol Q exogenous picked up by pinocytosis endocytosis and phagocytosis 0 betazmicroglobulin 3 2m invariant polypeptide that is common to all MHC Class I molecules also called the light chain of MHC Class I molecules 0 calnexin chaperone protein that keeps the heavy chain folded properly until 3 2microglobulin and peptide bind respectively 0 calreticulin aids in Tcell activation and composed of cytosolic serinethreonine phosphatase 0 chaperone protein along with calnexin 0 TAP transporter associated with antigen processing a heterodimeric protein that takes peptides from the proteasome and brings them into the ER proteasome in cytosol Q ERp57 enzyme associate the MHC class I heavy chain with TAP 0 tapasin associate the MHC class I heavy chain with TAP Q proteasome large multisubunit protease present in the cytosol of all cells that degrade cytoplasmic proteins generating the peptides presented by MHC Class I molecules 0 invariant chain covers the peptide binding groove so it is not loaded with endogenous peptides in the ER also shuttles Class II into endosomes 0 CLIP Class llassociated invariant chain peptide 0 invariant chain is digested by cathepsin 8 so only CLIP remains in peptide groove 0 HLADM releases CLIP and pops in a peptide from the endosome digest 2 For MHC Class and MHC class II identify which T cell coreceptor molecule they associate with MHC Class I gt CD8 cells MHC Class II gt CD4 cells 3 Identify the usual source endogenous exogenous of peptide that are presented in MHC Class land ll 0 MHC Class I carries peptide fragments from endogenous proteins proteins produced inside the cell in the cytosol Q MHC Class II carries peptide fragments from exogenous proteins proteins picked up by pinocytosis endocytosis phagocytosis Which of the following antigen presenting cells can only obtain antigen by receptor mediated endocytosis B Lymphocyte 4 Describe the general structure of the peptidebinding groove of MHC Class I and MHC Class II molecules and the size of peptide that each can hold 0 MHC Class I the at and d2 domains of the Class I heavy chain form the or helices second protein in dimer is invariant 32 microglobulin It can hold peptides that are 810 amino acids long 0 MHC Class II the at and 31 domains of the or chain and 3 chain respectively form the or helices It can hold peptides that are 1325 amino acids long 0 The open ends of MHC Class II allows bigger peptides 5 Describe how MHC Class I molecules traffic through a cell beginning at initial MHC protein synthesis and terminating at the cell surface including proteins that interact with the MHC protein as chaperones etc 0 Class I heavy chain is stabilized by calnexin until 3 2 microglobulin binds Q Calnexin is released and the heterodimer of class I heavy chain and 3 2M forms the peptide loading complex with calreticulin tapasin TAP ERp57 and PDI Q A peptide delivered by TAP binds to the class 1 heavy chain forming the mature MHC class I molecule 0 The class I molecule dissociates from the peptideloading complex and is exported from the endoplasmic reticulum 6 Describe how MHC Class II molecules traffic through a cell beginning at initial MHC protein synthesis and terminating at the cell surface including proteins that interact with the MHC protein as chaperones etc Carries things outside the cell When in the ER and theres peptide in the ER which is supposed to go into class one you have to plug up the MHC class 2 so it doesn t take up endogenous material MHC class 2 alpha and beta chains pair in the ER and are associated with invariant chain Invariant chain blocks binding of peptides to MHC class 2 molecules in the ER and shuttles class 2 into endosomes Once the MHC class 2 is in the endosome and leaves the ER proteases become activated In vesicles invariant chain is cleaved and digested by cathepsin S leaving the small peptide of CLIP fragment bound CLIP blocks binding of peptides in the MHC class 2 vesicles until peptides can bind HLADM facilitates release of CLIP allowing peptides to bind Then it is sent to the cell surface 7 Identify the body cells that do not express MHC Class I Erythrocytes are the only ones because they don t have a nucleus or a mitochondria Their only function is to carry oxygen These are also known as Red Blood cells 8 Identify the 3 professional antigenpresenting cells and describe how they obtain the antigen they present 0 Dendritic cells highly pinocytotic and phagocytic when immature can activate naive T lymphocytes 0 B lymphocytes present soluble antigens endocytosed bound to BCR Best antigen receptor 0 Macrophage present phagocytosed bacteria yeast etc 9 Briefly define what is meant by cross presentation and for which type of infectious agent bacterial viral fungal parasitic this process is necessary Q Is the special presentation of peptides within MHC class 1 to stimulate CD8 cytotoxic cell responses to viruses Exogenous antigen is getting placed into MHC 1 molecules pg 13 Topic 10 Major Histocompatibility Complex and HLA Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to if appropriate 0 HLA human leukocyte antigen clustered on chromosome 6 diverse genes sequence varies from person to person but not randomly Q Polygeny more than one gene encode MHC Class and Class II molecules HLA A B C all encode for class 1 for example 0 Polymorphism Hundreds of different alleles of the MHC genes that exist within the human population There is incompatibility from one persons tissues to another 0 Allogeneic Another person s allele that it antigenic and causes an alloreaction Q Alloreaction adaptive immune response made by one member of a species to an allogeneic antigen from another member of the same species 0 AlloantibodyH LA typing method Test serum for antibodies that could bind and kill cells from potential donor 0 GVHD graft vs host disease HLA mismatching leads to rejections of the graft This specific disease happens if it is a bone marrow transplant 2 Identify HLAA HLAB and HLAC as classical MHC Class I molecules and HLADP HLADQ and HLADR as classical MHC Class II molecules HLAA HLAB HLAC MHC Class I HLADP HLADQ HLADR MHC Class II The class 2 have 2 letters 3 Appreciate the relative numbers of alleles that exist for each HLA gene within the human population A person expresses all alleles from each parent Several hundred Ciro 239 viii s l 39a39 quotif333 Er r o g infrEEEI13L9rri 71 I A 506 B 372 MHC c 23 class ll E 3 F 4 G 10 DMA 4 DMB 7 DOA 3 DOB 4 D PA1 15 m DPB1 114 class ll Damquot 25 DQBl 66 DEA 2 DRB1 466 D RBS 3 D RBIlv I DRBS 15 4 Describe the inheritance of HLA haplotypes and the number of Class I and Class II isoforms that can compose an individual s haplotype A heterozygous person expresses two haplotypes Minimum 3 class I 3 class II alleles identical from each parent this is rare Maximum 6 class l8 class II alleles if both parents have DRE 5 Describe why transplantation of solid tissue or bone marrow cells from one human to another requires HLA typing and matching and the 2 possible adverse consequences of mismatches HLA mismatch can lead to graft rejection or graft versus host disease If bone marrow is not compatible with the recipient than the bone marrow will get attacked because it is the host and it s a painful death If typing is done incorrectly then you reject the graft and kill the organ by necrosis Typing is done by a combination of serology and molecular testing 6 Define what is meant by degenerate peptide binding in relation to MHC include an explanation of anchor residues HLA protection at the individual organism level stems from degenerate peptide binding to the groove MHC Class I 810 amino acid peptides MHC Class II 1325 amino acid peptides 2 or 3 of the amino acid residues in each peptide have to match the particular MHC protein groove expressed anchor residues compatible side chain reactions that allows ionic or polar bond to hold peptide in groove it allows ionic bonding or polar bonds you don t bind the entire protein only 23 amino acids which compose an anchor residue 7 Describe the currently known functions of HLAE and HLAG Both within HLA Class I region of chromosome HLAE binds peptide fragments to Class I leader peptides and presents to inhibitory receptors on NK cells Class 1 defines you as self send message to NK cells not to kill because you are self HLAG is restricted to fetal cells that lack HLA ABC and provides an NK cell ligand 8 Describe the relationship between interferon lFNalphabeta and changes in Class I expression They are used to upregulate the LMP2LMP7 proteasome subunits This allows them to preferentially cleave small VIRAL peptides to load into MHC class 1 Therefore this upregulates expression of MHC class 1 on cell surfaces increase class 1 expression on the cell and increase production of peptides into class 1 This allows an easy way to present viral peptides for immune system pg 14 Topic 11 B Lymphocyte Development Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to if appropriate PreBCR surrogate light chain RAG1RAG2 CAM cell adhesion molecule and polyspecific antibody 0 PreBCR lmmunoglobulinlike receptor that is expressed on the surface of proB cells It consists of umicro heavy chains in association with surrogate light chains formed from lambda5 and VpreB lts appearance signals the cessation of heavychain gene rearrangement Q Surrogate light chain Protein that mimics an immunoglobulin light chain It is made up of two subunits VpreB and lambda5 and is produced by proB cells Together with the micro heavy chain it forms the preBCR Q RAG1RAG2 Acronyms for recombinationactivating gene 1 and recombinationactivating gene 2 The proteins they encode are essential for the mechanism of receptorgene rearrangement in B cells and T cells 0 CAM cell adhesion molecule Along with early proB and Stem cells using the integrin VLA4 to bind to the adhesion molecule VCAM1 on stromal cells CAMs promote the binding of the receptor Kit on the B cell to stemcell factor SCF Q Polvsnecific antibodv Made against common bacterial polysaccharides and other carbohydrate antigens but are of little importance in making antibodies against protein antigens 2 Properly order the cell stages that occur during B cell development immature B cell naive mature B cell preB cell proB cell stem cell and identify characteristics of each that place it into that particular category eg preB cell has transient expression of preBCR rearranged lg heavy chain begins VJ rearrangement of light chain expresses VpreB A5 lga and lgB proteins 1 HSC In bone marrow gives rise to all progeny partially committed no embryonic stem cells 2 Lymphoid progenitor Can give rise to NK cell B cell or T cell 3 ProB Cell Begins rearrangement of heavy locus earliest identifiable B lineage cell Heavy chain DJ to VDJ rearrangement no immunoglobulin status 0 Express cKit lL7 and VLA4 lgM gets made from ProB cell to PreB cell 4 PreB cells that make a productive heavy chain rearrangement locus transient expression of preBCB rearranged Ig heavy chain begins VJ rearrangement of light chain 0 Expresses VpreB lambda5 lga and lgb proteins signals and causes burst of proliferation 5 Immature B Have surface expression of productively rearranged u heavy plus rearranged light chain surface lgM essentially mature B cell receptor in form lgM 6 Mature B are ready to see antigens now F 3 Explain why it is advantageous for a preB cell to undergo a burst of proliferation following signaling through the preBCR 0 Quantity to get as many developing preB cells out there as possible 0 Diversity To get the quantities up and get as many Pre B cells out there as you can get diversity Q Efficiencv of effort Efficiency of effort is the advantage because when somatic recombination occurs it only has 13 a chance of getting the sequence right each time it is spliced together 0 Getting the numbers out there 0 You want to replicate the good heavy chain so it is produced more You don t lose anything because the heavy chains get paired with different light chains every time so the antigen specificity of the pool will still be diverse 4 List when RAG1 and RAG2 are expressed during B cell development ie what stages 0 RAG1 and BAG2 are transiently expressed in ProB and PreB cells developing T cells and B cells only during the time the heavy chains and the light chain rearrangements are being made 0 Presence of preBCB or BCR on cell surface turns off gene expression 5 Identify the role of stromal cells in B cell development and where these stromal cells are located in the body 0 Bone marrow stromal cells are nonlymphoid lineage cells that allow for B cell development 6 List the 2 growth factors that support B cell development in the bone marrow 0 Bone marrow stromal cells provide two important signals for developing B cells 0 Cellcell contact via cell adhesion molecules CAM s O Membranebound growth factor stem cell factor SCF and soluble cytokine growth factor interleukin7 IL7 7 Identify what percent of developing B lymphocytes successfully make it through each major step of somatic recombination during development heavy chain rearrangement light chain rearrangement 0 Heavy chain gene rearrangement 23 0 Light chain gene rearrangement 85 o F B Cells Must Malia 39 Ftr uctive Heavy and Light Ehai Rearrangements t Survive ll Elsi arr a g r Ent 7 39539 DJ r Frrang mm Pi urn bandit urban fE fa En t ihr l w E 1 Fltmarriages LE giant EH1 Erin I39ImnFi39iaas r 1139 EJl r39 Fran57mm V an scamuntil absum s rHE 39 7 v39 39 l H H FEEBanning againE r m nd hfl Hananages Luggage ii an rst Ei E E E r xi X s 39 39 39 quot Wmiilwmm EEMEM J FEHirrangahgtnt l 53 1H1 LE n d charge quotu P W H l ljli39i dmii 39i mummyLemur i l i d ii H wwaJ zTMm qimMH ig i Wim lPT dlUrEEl39UE f in39lFFElml rearrangement E i li rl is 3 HUEIKE LidESJ sit irin 1 are praductve T 8 Explain why lg light chain rearrangement has a higher rate of success than lg heavy chain rearrangement Q The success rate in obtaining a functional lightchain gene rearrangement is much higher than that for the heavychain gene because 0 Only two gene segments are involved compared with three for the heavychain gene 0 The B cell has four lightchain genes two kappa and two lambda with which to work two types of loci 0 Light chain rearrangement only requires the formation of VJ whereas heavy chain rearrangement requires the formation of VDJ 0 Every B cell has two copies of the Ig heavy chain locus Two copies of each are on homologous chromosome one from mother and one from father so there are 2 chances to try VDJ rearrangement 9 Identify the two CD markers that when used in combination can identify B1 B lymphocytes note need to differentiate B cell from T cell 0 B1 CD19 CD20 and CD5 0 B2 D19 CD20 Q T CD5 identifies cell as a B1 cells as opposed to a B2 cell 10 Compare and contrast B1 and 32 B cells in regards to which is produced primarily in a fetus selfrenewal capacity spontaneous lg production major isotype secreted relative level of somatic hypermutation and ability to respond to carbohydrate and protein antigens B1 cells 32 cells When first produced Fetus After birth Mode of renewal Selfrenewing Replaced from bone marrow Spontaneous lgM High LOW pl OdUCtIOI I Major isotype secreted lgMgtgtgtlgG lgGgtlgM Somatic hypermutation Lownone High Ability to respond to carbohydrate and protein Carbohydrates Proteins an gens pg 15 Topic 12 B Lymphocyte Apoptosis or Activation Objectives At the completion of the module and the assigned reading the student should be able to 1 Define the following terms including what ces the term applies to if appropriate 0 Clonal deletion If an immature B cell s BCR lgM is crosslinked by self antigen on cell surfaces and cannot remove the selfreactivity by receptor editing the B cell is induced to commit suicide by apoptosis The BCR is all pulled to the same side of the cell and is maximally activated Solid Phase selfAg 0 Clonal anergy Immature B cells that bind soluble selfantigen are not deleted but become anergic Soluble selfAg Q Anergic nonfunctional immune cells nonfunctional if you can t get rid of selfreactive B cell 0 Selftolerance the normal situation whereby a person s immune system does not respond to constituents of that person s body the circulating lymphocyte population in an individual is thus selftolerant 0 LT lymphotoxin subset of TNFfamily cytokines involved in development and maintenance of the architecture of secondary lymphoid tissues and organs 0 BAFF Activating Factor cytokine that belong to the tumor necrosis factor ligand family Potent B Cell activator Involved in proliferation and differentiation of B cells 0 Primary lymphoid follicle Secondary lymphoid organs B cells congregate here and interact with the follicular dendritic cells 0 Secondary lymphoid follicle a primary follicle in which activated B cells form a germinal center 0 Germinal center area in secondary lymphoid tissue that is a site of intense Bcell proliferation selection maturation and death 0 Plasma cell also called plasma B cells plasmocytes and effector B cells are white blood cells that secrete large volumes of antibodies 0 Centroblast generally refers to an activated B cell that is enlarged and proliferating encompassing both large and small noncleaved follicular center cells 0 Centrocyte generally refers to a B cell with a cleaved nucleus as may appear in eg follicular lvmphoma During B cell development centrocytes are formed following the cessation of centroblast proliferation 2 Define receptor editing and whether it is allowed on heavy chains light chains or both 0 Receptor Editing occurs when a Bcell in the Bone marrow is reactive to self cells 0 It tries to rearrange and change the antigen binding site RAG genes are turned back on and light chain rearrangement resumes Heavy chain NOT edited Successive edits can be tried lf unsuccessful die by apoptosis If successful leave bone marrow pass negative selection Heavy chain can receptor edit but not in response to selfantigen 3 State the average lifespan of a B lymphocyte after it exits the bone marrow and enters circulation Q B cells have a halflife of 100 days if they do not encounter their antigen and become activated 4 Trace the path of a B lymphocyte from entry to exit of a lymph node assuming it does NOT become stimulated by antigen in the lymph node 1 B cell enters via afferent blood vessels 2 squeezes through HEV high endothelial venules into the T cell area of the cortex 3 If not activated it migrates out of T cell area to primary follicle in B cell area 4 then exits via efferent lymphatic vessel 5 Identify the cell that provides necessary periodic survival signals to a naive B lymphocyte where this cell is located and what molecule it produces that provides the survival signal 0 Follicular dendritic cells provide survival signals for the B cells 0 located in the Primary Follicle of lymph node 0 BAFF from FDC provides lifesaving maturation signal to B cell 0 LT from B cells provide life signal for FDC 6 Explain what cell interaction triggers isotype switching and somatic hypermutation Q TCell Interaction in Lymph node 0 Activated B Cell presents antigen to MHC class II to CD4 TCell which causes T Cell to secrete cytokines which influences B Cell Causes changes in isotype to most useful isoptype of antibody for type of antigen being exposed at the moment T Cell triggers somatic hypermutation in B Cell and T Cell influences whether these B cells develop in germinal center where clonal selectionaffinity maturation occur or BCells drop out right away and become lgM secreting plasma cells in medulla of lymph node Q The B cell binds antigen via lg BCR endocytosis and processes into MHC II and presents peptide to CD4T cell that responds to that same antigen not same epitope This is necessary to ensure T cell help is directed to the proper Agmatched B cell B cell makes it to LN and finds a T cell that recognizes same antigen they connect that lockandkey is linked recognition the T cell sends cytokines over to determine the fate of the B cell 0 Don t want T Cell to help self activated B Cell 7 ELIth 39 quotcan b 030th an 31 3quot n p iv at A Al u t it it b w lea t wat39 V t 1 cm itquotsL gggeuct am focal ti Th elm Ul 1 15quot gait Tin an cLMm t Fatima LII 11H 6 QM his w m 113 Ln E3 5 quot39w Mute Emit 7 77 39 mh m i MT MM H i 911 1 quot 8 Identify the main function of a plasma cell and the 4 locations where it may reside in the body 0 To secrete as much antibody as fast as it can 0 The four locations where it may reside in the body are in the O lymph node medulla O spleen red pulp 0 bone marrow O GALT 9 List the 3 main options for what a B lymphocyte can do functionally what it can become m it is activated by antigen 0 Can become large proliferating centroblasts which mature into small non proliferating centrocytes 0 Some differentiate to plasma cells and leave the germinal center 0 Some develop into resting memory B cells pg 16 Topic 13 T Cell Development Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms doublenegative thymocyte doublepositive thymocyte singlepositive T cell pTu Q Doublenegative thymocyte O No CD4 or CD8 0 Most immature T cell in thymus 0 These progenitors still have the TCR genes in germline configuration no somatic rearrangement yet 0 These progenitors can give rise to both dB and V6 T cell Hneages Q Doublepositive thymocyte O T cell in an intermediate stage of development in the thymus lt expresses both CD4 and CD8 Q Singlepositive T cell 0 Final stage is to CD4CD8 or CD4CD8 singlepositive T cells or mature T cells a late stage of T cell development in the thymus and characterized by the expression of either CD4 or CD8 coreceptor on the cell surface 0 pTor O The surrogate or chain that combines with the Tcell receptor 3 chain to form the preTcell receptor 2 Identify what receptorligand interaction triggers commitment of an uncommitted lymphoid progenitor to the T lymphocyte lineage and where this interaction occurs 0 Notch1 receptor on progenitor CLP binding to Delta1 ligand expressed by thymic endothelial cells in thymus commits the progenitor to T cell lineage 3 Identify the cortex and medulla of the thymus and what cell lineages can be found in each 0 Two cell types compose the shell of the thymus O l thymocytes and macrophages O l dendritic cells macrophages and thymocytes Q Progenitor cells from bone marrow populate the thymic structure 0 Thymocytes from CD34 progenitor immature developing T cells 0 Thymic dendritic cells from CD34 progenitor in medulla O Macrophages of bone marrow origin concentrate in medulla and scattered in cortex 0 Cortex O Cortical epithelial cells thymocytes and macrophages1 Q Medulla O Medullary epithelial cells dendritic cells macrophages and thymocytes 4 Define thymic involution and when it occurs 0 The thymus atrophies and fills with fat as we age called thymic involution 0 Therefore majority of T cell repertoire is determined by age 30 O Thymic involution does not appear to hinder T cell immunity 0 Thymic involution is when the thymus atrophies and this typically occurs around age 30 where the T cell pool is completed Although it occurs early in life thymic involution does not affect T cell immunity thus naive T lymphocytes live much longer than naive B lymphocytes QUESTION 5 List the proteins that compose the preTCR Q PreTCR TCRB pTor CD3 complex CD4 CD8 Cchain 0 Two zeta chains CD3 complex consisting of a and 6 chains y and a chains along with a pTor 6 Identify whether a preT cell would be a doublenegative thymocyte a doublepositive thymocyte or a single positive T cell 0 Double positive thymocyte CD4CD8 preT cell 0 A doublenegative thymocyte has yet to turn on CD4 or CD8 Double positive thymocytes express CD4 and CD8 but have not decided on which it will ultimately express Singlepositive thymocytes have either CD4 or CD8 0 double positivepreTcell 7 Describe the order of rearrangement of the or B 6 and y TCR gene loci during thymocyte development and how a T cell becomes either an dB or V6 T cell 1 Tcell precursor that enters the thymus uncommitted progenitor expresses the CD34 marker 2 Once it commits to become T cell lineage it expresses CD2 and CD5 and becomes a doublenegative Tcell progenitor 3 Begin rearranging By amp 6 gene loci simultaneously 4 If the cell can express the y 6 receptor on its surface before 3 chain rearranges it will become a committed y6 T cell 5 If the cell rearranges 3 chain first it will show up on the surface with a surrogate alpha chain called pTd high proliferation 6 Now the second round of somatic recombination begin rearranging or but continue to rearrange y and 6 still possible for cell to become committed y 6 T cell 8 Identify the normal proportion of dB to V6 T cells produced after birth 0 gt90 o become dB T cells 0 10 gamma dellta pg 17 Topic 14 T Lymphocyte Selection Objectives At the completion of the modules and the assigned reading the student should be able to 1 Identify whether B cells alphabeta T cells and gammadelta T cells each undergo positive andor negative selection 0 B cells undergo only negative selection 0 alphabeta T cells undergo both positive and negative selection 0 gammadelta T cells do not undergo neither positive nor negative selection do not see antigen in MHC Class or 0 although there are some differences 0 For B cells clonal anergy is in response to soluble antigen 0 For T cells selfantigen within MHC in the periphery cell bound not soluble O 109 97 2 Describe the purpose of positive selection and negative selection 0 Positive selection The purpose of positive selection is so that only useful Tcells that are able to bind self MHC are released into the periphery Negative selection The purpose of negative selection is to weed out the Tcells that are reactive to self antigens 0 v6 T cells do not undergo negative selection 3 Identify what cells mediate positive selection and negative selection of T cells 0 Positive selection inner cortex cortical epithelial cells amp thymocytes of bone marrow origin 0 Negative selection medulla dendriticamp macrophages 4 Describe the function of dendritic cells and macrophages in the thymus note they are different 0 dendritic cells work with negative selection 0 Both are good at Presenting antigen to T cells 0 Macrophages good at phagocytosis 99 of T cells fail Pos amp Neg selection 0 Macrophages are removing that debris 5 Identify the AIRE protein what cell expresses it and whether it is important in positive or negative selection Q AIRE autoimmune regulatory protein is a general transcription factor and induces production of many tissuespecific cellular proteins at low levels in the thymus 0 important for negative selection in the medulla to avoid autoimmune disease 0 medullary epithelial cells express 6 Describe how strength of TCR binding is important for positive and negative selection Q If TCR binding is too avidlytight the self MHC will fail negative selection and undergo apoptosis 0 Must be able to bind moderately for positive selection If it binds too tightly negative selection by apoptosis 0 T cells that are positively selected must be able to bind to either MHC class I or class II containing selfpeptides expressed on cortical epithelial cells 7 Describe the difference between central tolerance and peripheral tolerance in regards to T cells and whether each is achieved by either anergy or apoptosis 0 Central tolerance Prevents autoimmunity deletion of selfreactive T lymphocytes in thymus by negative selection induced by apoptosis in primary lymph organs 0 Peripheral tolerance Controls autoimmunity Treg cell activation control anergy induction 8 Define Treg and describe their cellular characteristics and the role they play in tolerance Q T reg are subset of CD4 alphabeta T cells 0 express CD25 lL2Ralpha O express FoxP3 transcription factor Q secrete cytokines IL10 andor TGFbeta that suppress normal T cell activation 0 multiple subsets of Treg appear to exist I central Treg that arise in the thymus l iTreg induced T reg that are educated and differentiate in secondary lymphoid tissue in particular in the GI tract pg 18 Topic 15 T Lymphocyte Activation Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to humoral immunity cellmediated immunity costimulatory molecule B7 CD28 twosignal hypothesis CTLA4 Q Humoral immunitv Antibody immunity CD4 T helper 2 cell 0 Cell mediated immunitv CD4 T helper 1 Q Costimulatorv molecule Helps activate naive Tcell delivered only by professional APC dendritic cells macrophages and B cells The costimulatory molecule on APC is B7 which binds to CD28 on the T cell 0 E On APC costimulatory molecules Binds CD28 or CTLA4 0 CD28 Cell surface protein on naive Tcell Q CTLA4 Inhibitory receptor helps terminate T cell immune response 0 limits duration of response 0 lack of CTLA4 can lead to lymphoproliferative disorders 0 variants of this can lead to autoimmunity 2 Identify the 3 main effector T cells and the important role that each has in the immune system 1 CD 8 Cytotoxic T cells killer T cells 2 CD 4 T helper 1 cell Cell mediated immunity 0 Assist macrophages and cytotoxic T cells 3 CD 4 T helper 2 cell Humoral immunity Antibody immunity 0 Aid B cells to aid antibody immunity aka humoral immunity 3 Describe the two ways that foreign antigen can reach the secondary lymphoid tissues to activate an adaptive immune response 1 Blood capillary HEV 2 Afferent lymph vessel from upstream LN 4 Identify the general role of selectins and integrins in lymphocyte trafficking Q Selectins provide low affinity adhesion that causes cells to roll along blood vessel walls within HEV or activated endothelium of inflamed tissues they bind to vascular addressins which are glycoproteins on endothelium that bind selectins Selectins SLOW THEM DOWN Q lntegrins provides highstrength high affinity binding that allows firm attachment and squeezing between endothelial cells diapedesis and extend contact between lymphocytes and APC they are heterodimers ex LFA1 They bind to Some lglike superfamily proteins which are cell adhesion molecules present on lymphocytes dendritic cells and activated endothelium and bind intergrins ICAM S 5 Identify the three functions that leukocyte adhesion molecules must perform prior to successful activation of a nai39ve T lymphocyte by an APC O Naive T cells Lselectins target CD34 GlyCAMt which are part of the HEV MAdCAMtmucosal endothelium O Naive T cells LFA1 targets l lCAM1 and lCAM2 high affinity O T cell LFA1 lCAM3 CD2 targets lCAM1 and lCAM2 LFA1 and DCSIGN LFA3 6 Identify the primary antigen type bacteria yeast virus soluble protein that DC macrophages and B cells specialize in presenting 0 DC Virus 0 Macrophages Bacteria Yeast Q B cells Soluble protein Toxins Viruses 7 Define the twosignal hypothesis and identify the ligandreceptor proteins that provide the second signal including specifically what cells these ligands and receptors are expressed on Q The twosignal hypothesis is the primary signal that provides MHC with antigen peptide groove that triggers the TCR and has to provide a costimulatory signal which maintains and amplifies autocrine lL2 production that is necessary for T cell activation The costimulatory molecule B7 on the APC binds to CD28 on naive T cell 8 Describe the importance of lL2 in T cell responses and how the 3 immunosuppressive drugs Cyclosporin A FK506 and Rapamycin shortcircuit this signal 0 lL2 prevents T Cell from becoming Anergic O Cyclosporin A and FK506 inhibits lL2 production by naive T cells 0 Rapamycin inhibits signaling by lL2 Receptors 0 Together they can help prevent transplant rejection 9 Discuss what factors minimally the 4 mentioned in class can influence the choice of a nai39ve CD4 T cell to a functional subset choice Th1 Th2L and which of these factors appears dominant Q T cell subset choice is influenced by O Cytokines in vicinity most dominant 0 APC costimulatory molecules 0 Concentration of Ag low conc favors Th1 high conc favors Th2 O TCR affinity low Th2 high Th1 immune response is mixture of phenotype and dominant subset is one that is best counter to the pathogen 10 What cytokines do Th1 Th2 and Treg secrete and what transcription factors are responsible for activating this secretion in each cell type 0 Treg secretes TGFbeta and lL1O and the trans factor is Fox P3 0 TH1 secretes lL2 and lFNgamma and the trans factor is Tbet Q TH2 secretes lL4 and lL5 and the trans factor is GATA3 11 Identify what cytokine is needed in high concentration to activate a nai39ve CD8 Tcell to a Tc and which cell can aid the APC in this activation process 0 lL2 is needed in high concentration 0 Dendritic cells with high costimulatory molecule expression trigger optimal lL2 response B cells and macrophages can t do this 0 T helper CD4 cells can also activate APC to express necessary co stimulator molecules to activate CD8 cells directly SIDE ADDITIONAL CLASS QUESTIONS 0 Draw out the cells and moleculesreceptors involved in linked recognition and the twosignal hypothesis 0 Which involves naive T cells and which one involves activated T cells 41 4 w w l My an Vw W A m i t 1 ll i J J a Ra 3 430 us Iquot J Hawk pg 19 Topic 16 T Lymphocyte Functions Objectives At the completion of the modules and the assigned reading the student should be able to 1 Identify where Th1 Th2 and To reside following activation in the secondary lymphoid tissue 0 Th1 MOST exit the LN and travel to sites of infection 0 Th2 STAY in LN tissues to aid B cells in their response 0 To ALL exit the LN tissues and travel to sites of infection 2 Describe how adhesion molecules akahoming receptors differ on effector T lymphocytes compared to nai39ve T lymphocytes 0 Effectors T cells change adhesion molecule expression to broaden cells they can interact with instead of primarily APC Q Decrease in Lselection which is what a naive T cell uses to roll in high endothelium venules and increase VLA4 to home to inflamed blood vessel endothelium instead of HEV of secondary lymphoid tissues 0 Increase CD2 and increase LFA1 to bind target cells expressing low levels of their ligands LFA3 and lCAM1 3 Identify the functions of Fas ligand FasL and CD40 ligand CD4OL in regards to effector T cell functions 0 FasL lnduces apoptosis of any target cell expressing Fas Q CD4OL T helper cell uses to activate Bcells or macrophages 0 B cell requires this signal to stay alive 4 Identify the three major proteins found in To lytic granules and the functions of each protein 0 Granulysin Cooperates with perforin to create a transmembrane pore Q Perforin Related to C9 cooperates with granulysin to form a transmembrane pore Q Granzymes Enter through said pore and cleaves proenzymes and caspase sp in the target cell 0 Activates apoptosis 5 For the following cytokines identify the cells that produces it and its main func on Cytotoxic T Cells Cytokines lFNg LT TH1 Cy5tokines lFNg GMCSF TNFa LT lL3 TH2 Cytokines lL4 lL5 lL10 lL13 TGFb Q lL2 Q from activated T cells 0 essential in proliferation of activated T cells and development of adaptive immune response 0 lL4 Q from Th2 cells to activate B cells 0 lL5 Q from Th2 cells to activate B cells 0 lL6 0 released by macrophages 0 with lL2 and TNF or induces a wide range of inflammatory responses at early times in infection 0 lL1O O inhibits Th1 by decreasing MHC expression on APC Q lFNgamma 0 released by both To and Th1 to activate macrophages 0 GMCSF 0 released by Th1 cells to increase myelopoiesis neutrophils amp macrophages in bone marrow Q TNFalpha 0 released by both To and Th1 to activate macrophages 0 activate vascular endothelium at site of infection to increase phagocyte recruitment 0 LT O activate vascular endothelium at site of infection to increase phagocyte recruitment Q TGFbeta 0 an immunosuppressive cytokine released by Treg cells and other cells that functions to decrease macrophage activation Main Function Cytokine Produced By lL2 Activated T cells Essential in proliferation of activated T cells and development of adaptive immune response lL4 Th2 cells To activate B cells lL5 Th2 cells To activate B cells lL6 Released by macrophages With lL2 and TNF or induces a wide range of inflammatory responses at early times in infection lL1O Th2 cells Inhibits Th1 by decreasing MHC expression on APC lFNy Released by both To and To activate macrophages Th1 Inhibits TH2 responses GMCSF Released by Th1 cells To increase myelopoiesis neutrophils amp macrophages in bone marrow TNFor Released by both To and 0 To activate Th1 macrophages 0 To activate vascular endothelium at site of infection to increase phagocyte recruitment LT To activate vascular endothelium at site of infection to increase phagocyte recruitment TGFB lmmunosuppressive Functions to decrease cytokine released by Treg macrophage activation cells and others 6 Describe how interferons lFN s lFNor lFNB lFNy are especially important to fight viral infections 0 Inhibits replication of viruses in infected cells 0 Increases processing and presentation of viral antigens in Class by increasing LMP2LMP7 in proteasome to cleave smaller peptides Q Activates bystander macrophages to phagocytize apoptotic cells 0 lnterferons cytokines that help cells to resist viral infection 0 lnterferons tVDe 1 includes IFN or and IFN 3 produced by leukocytes and fibroblasts act specifically to induce cells to resist viral infection 0 y product of CD4 Th1 cells CD8 T cells and NK cells has more general functions in immune responses acting principally to activate macrophages Describe the features of a granuloma and the circumstances under which a granuloma may form 0 Granuloma is mass with an organism Pathogen in the middle Q It is walled off by fused macrophages 0 it is surrounded by single macrophages on the outside of that 0 Outside of that it is encircled by T cells 0 Granuloma is created because pathogens live in macrophages in vesicles and cannot be destroyed by them This leads to chronic infection 8 List the 3 main cells that accomplish cellmediated immunity 0 Macrophages Q Th1 0 Tc pg 20 Topic 17 B Lymphocyte Activation Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to as appropriate DC FDC and immune complexes 2 List the 3 CD proteins that form the B cell coreceptor complex and the functions of each of the proteins if known 0 CD19 increase BCR signal 100010000 fold 0 CD21 also known as complement receptor 2 ligands are ic3b or c3d these ligands cause an additional signal to go thru CD19 to increase BCR signal 1000 10000 fold 0 CD81 no function known multispanning protein 3 Compare and contrast Tdependent and Tindependent B lymphocyte antibody responses Be sure to identify the subsequent characteristics of these two responses in regards to the main antibody isotype produced whether isotype switching andor affinity maturation occur whether the B cell activation is antigenspecific vs polyspecific and whether a memory response is induced T dependent refer to B cell antigens that require T cell help for proper response antigen specific 0 Respond to protein antigens 0 Usually takes 4 cells 0 Activation of B cells for production of TD antibody responses occurs in the germinal 0 High affinity o isotypeswitched 0 require activation of B cell in germinal center T cell help and provide memory Tindependent Cause B cell responses in the absence of T cell help o Antibody produced in response by the B cell is generally low affinity no somatic hypermutation 0 Minimal isotype switching mostly lgM isotype o Antigen is commonly a carbohydrate o No protective memory response produced center and requires linked recognition of antigen by CD4 T helper cell hypermutation ANTIBODY PRODUCED IS POLYSPECIFIC 4 Briefly explain how TH and Tl2 antigens differ 0 TH Antigens trigger B cell activation while using TLR Signals Pattern recognition receptors P Q Tl2 antigens are extremely repetitive antigens on pathogen surfaces doesn t require a TLR signal 5 List the 4 cell types that are minimally required to allow a Tdependent antibody response to occur in a secondary lymphoid tissue and the relative physical location of each cell within the secondary lymphoid tissue 0 Dendritic cell T cell B cell Follicular dendritic cell 0 The initial activation of the T cell its differentiation to T helper cell then the T helper cell forming the antigen cognate with the B cell all occur in the T cell area or the cortex of the lymph node 0 The B cell interacting with FDC then B cell at T helper cell happens in the germinal center where it forms a secondary follicle inside of a primary lymphoid follicle DC Ag T gt Th B gt BTh FDCICAg 0 blue interact in the T cell area cortex of LN 0 purple interact in germinal center of 2ndary follicle 6 List the cells that are normally located in the dark zone light zone and mantle zone of a germinal center include centroblasts centrocytes FDC and T helper cells in your list centrblasts 0 Dark zone centroblasts B cells 0 Cells that are proliferating madly pushed together closely 0 Light zone Centrocytes present Ag to Th in the outer region of the light zone FDCs loosely interact with B cells and CD4 helper T cells in the light zone of the germinal center 0 Outer zone of centrocytes that are undergoing selection process 0 Mantle zone contains unactivated B cells that do not recognize the specific Ag and got pushed out ofthe way 0 Surrounds light zone 7 Identify as many important functions of FDC as you can and predict the ultimate effect on the immune system if FDC were absent from secondary lymphoid tissues 0 They are Stromal Cells 0 Different from regular Dendritic cells 0 They bind Immune Complexes AgAb and hold them on surface for extended penod 0 Hold them as lccosomes Q Loosely interact with B cells and CD4 T cells in the light zone of a germinal center 8 Identify the cytokine that is required for differentiation of FDC o W and LTBgt 0 Without LT the follicles don39t form 9 Explain the 2 signals a B cell requires in a germinal center to maintain activation and the consequence if it does not receive these signals 0 They need the binding of the antigen held on the FDC 0 They B cells need to process the antigen and then present it to CD4 Tcells O T Cell provides second signal via CD40 which induces BclXLcrucial to preventing Apoptosis Q linked recognition cognate interaction 0 If these 2 are not given then cells die via apoptosis 10 Identify the cellsurface molecule on a T helper cell that is absolutely critical for driving Tdependent antibody responses CD40 pg 21 Topic 18 Immune Functions of Antibody Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to as appropriate 0 Fix complement Binding of compliment which is initiated by the pentamer lgM Q Polvlg receptor lg receptor Bcells in MALT produce lgA as a dimer which has a J chain of dimeric lgA that is bound by a polylg receptor on basolateral surface of mucosal epithelial cell and released on the apical face with a fragment of the polyig receptor now called secretory component still a ached Q Jchain Two identical joining monomers of lgA held together with a J chain joining Q Secretorv Component See polylg receptor above 0 E fragment crystallizable lgG is bound via Fc region to FcRn within endocytic vesicles of capillary endothelium following pinocytosis Q FcRn transports lgG across endothelial cell and releases into extracellular spaces occurs constitutively in the absence of inflammation 0 FcR Fc receptors on phagocytosis bind antibodycoated pathogens opsonization 2 Define and provide examples of the major functions of antibody including neutralization opsonization complement fixation and ADCC Q Neutralization Neutralize toxins andor pathogen binding to cells 0 Example 0 Opsonization Opsonize target for phagocytosis 0 Example 0 Complement Fixation 0 activation for I For direct lysis of bacteria l Phagocytosis by myeloid lineage cells via CR complement receptor l Clearance of immune complexes by CR1 on RBC l Example 0 ADCC Antibody Dependent Cellular Cytotoxicity Tagging infected cells for direct lysis by NK cells 0 Example 3 Identify the 3 major antibody isotypes in body fluids and specifically indicate whether each isotype is confined to the bloodstream or can enter tissue fluids 0 lgM circulates in the blood with little access to tissues 0 lgG circulates in the blood and can diffuse into tissue fluids to encounter an gen O actively transported across blood vessel endothelium Q lgA circulates in the blood and can diffuse into tissue fluids to encounter an gen O actively transported across mucosal epithelium 4 Identify the protein that is necessary to produce lgA as a dimer 0 Two identical monomers of lgA held together with a Jchain joining Q Jchain of dimeric A is bound by polylg receptor on basolateral surface of mucosal epithelial cell and released on the apical face with a fragment of the polylg receptor now called secretory component still attached 5 Describe how lgA produced in mucosal lamina propria can get across the mucosal epithelial layer to be released in high concentration into mucus Binding of lgA to receptor on basolateral face of epithelial cell Receptormediated endocytosis of lgA Transport of lgA to apical face of epithelial cell Receptor is cleaved lgA is bound to mucus through the secretory piece 6 Describe the important functions of FcRn Brambell receptor in both pre and post natal human 0 FcRn transports lgG across endothelial cell and releases into extracellular spaces occurs constitutively in the absence of inflammation 0 Prenatal lgG needs Fc receptor to cross placenta neutralizeop Q Postnatal fight extracellular infection 7 Identify the two forms of passive antibody immunity that a fetusnewborn receives from its mother 0 Dimeric lgA Concentrated in breast milk especially colostrum and provides protection against gut bacteria 0 lgG Transfer across placenta protects infants from other infectious agents until baby s immune system is able to respond 8 Describe the relative order and time of appreciable appearance 3050 of adult levels of lgA lgG and lgM in the bloodstream during newborn early childhood years for antibody produced by the child itself not from mother 0 Before birth the mother passively transferred lgG increases and a baby starts synthesizing its own while the mothers decreases and at 3 months they are at the same level and the new lgG increases 0 lgM starts off very slow and after birth takes off 0 lgA doesn39t start until after birth and is slow to increase 9 Appropriately match each Fc receptor in the following list with its primary function function should be either opsonization inhibitory granule exocytosis or ADCC Opsonization Inhibitory Efgggt gseis ADCC FcorRt Fcle FclelA FclelBt FclelBZ chRl Fclell 10 Describe the role of BBC in removing immune complexes from the plasma 0 Immune complexes Toxin float around in the blood plasma 0 Cfg initiates classical complement pathway when it binds to the immune complex 0 The Immune complex is tagged and covered by C3b Covalently bound 0 RBC with CR1 receptors on surface binds to the Q and carries the immune complex with it through the bloodstream 0 RBC arrives at liver or spleen where immune complex is detached from the RBC aka an erythrocyte and is engulfed by a macrophage and digested pg 22 Topic 19 Mucosal Immunity Objectives At the completion of the modules and the assigned reading the student should be able to 1 Describe the organization of Peyer s patches being sure to identify abundant cell types within them and the function of the M cells Peyer s patches are in the small intestine they have a T cell area B follicles germinal center organized like a lymph node Has M cellmircofold which functions to serve lymphoid tissue epithelial origin 0 M cells take up antigen by phagocytosis and endocytosis Q Antigen is transported across M cells in vesicles and released on the basal surface 0 Antigen is bound by dendritic cells which activate T cells 0 Constantly transporting normal flora through the immune system so that B and T cells can respond on a regulatory basis 0 M cells are specialized cells with a ruffled surface They do not secrete mucus or possess overlying glycocalyx They have weak Iysosomes do not digest what they take up 0 MAIN FUNCTION IS TO SAMPLE GUT BACTERIA Underlying mucosa but separated from normal villi You can count them they are large Has lymphatic drainage that initiates in the lamina propria in the villi and peyers patches 2 Associate intraepithelial lymphocytes IEL with the gut and describe what their characteristics are and how abundant they are They re dispersed in the epithelial layer CD8 cytotoxic cells loaded with granules ready to kill 0 Prevents massive destruction of mucosal surface Mix of alpha beta and gamma delta TCR but with limited specificity o Populate during embryonic development One lEL for every 710 epithelial cells abundant 3 Identify chemokine and integrin receptors expressed differentially on IEL lamina propria lymphocytes and skin lymphocytes that are responsible for these cells homing to the correct body site 0 IEL O CCR9 CCL25 from gut epithelium O orEB7 integrin which binds to E cadherin of intestinal epithelial cells 0 Lamina Propria Lymphocytes B amp T O CCR9 and d4B7 integrin that binds to MAdCAMf mucosal addressin 0 Skin Lymphocytes O CCR4 and 04B1 integrin 4 List the three known cytokines that cause plasma cells located in the gut to isotype switch to lgA production APRIL TGFbeta lL1O 5 Identify the two main functions of dimeric lgA in the mucosa Neutralization Q Can neutralize antigens internalized in endosomes neutralize pathogens and toxins Transport 0 lgA can export toxins and pathogens from lamina propria while being secreted Q Secreted lgA binds pathogen on M cell surface and takes it to lymphoid tissue 0 Secreted lgA picks up antigen in the endosomes of the M cell and takes it to lymphoid tissue Dimeric lA functions primarily for neutraliztion and transport epithelial EII 6 Describe how the immune system seems to compensate for a lack of lgA in Selective lgA Deficiency Q It compensates by overproducing lgM and exporting lgM across mucosa Jchain via polylg receptor 0 Could be due to the fact that lgM fixes complement 7 Describe how Th2 responses in mucosal tissues are specifically effective at ridding the body of helminth infections including the specific functions of Th2 cytokines lL4 lL5 and lL13 Q Th1 functions are bad cause damage 0 Th2 functions help lL4 drives B cells to produce lgE lL5 recruit and activates eosinophils lL13 induces epithelial cell repair and mucus it also helps drive mast cell recruitment along with lL9 Specific lgE arms mast cells against helminths mast cells produce mediators such as histamine TNFalpha and MMCP These recruit inflammatory cells and remodel the mucosa o Increases mucus production by producing more goblet cells Activates eosinophils activates mast cell and lgE production lgE crosses Fc receptors on mast cells and causes them to degranulate This causes massive outflow of fluid across the gut fluid and increased production of mucus In developin countries gut helminths inuce Th2 responses u 39 f HtLJs V 7 HillIla 39 i9 it ath 39 fat 5 Elquot 51g lym 1 w are r 1 E v quot 39n 3939r lanesed tell Euslneplhllls lgE aim15 mist I eellls paraf n5e Fmdlu i elf itemgllement humeeer and pmdlmte HEP tells em 39fltdiatr f iiiEH15 fli ll d xing emlilmdes malignantlit help whith kills eaglemph to limitfirm Willii t39i j 39 phj gl i shedding at penmimTIiey anemiala gramme and M 39 mm new Em1 ms hed em Ilse antigen mnui damage errid niftllfze lial eellle HIDE using a in a mmlmry falls ti ue remndaling Mucus prevents parasiteapeiz c and remndel the adheremze and lg mama accelerates less at mm ie Figure ll l The Immune estami edt 21 Garland Etienne pg 23 Topic 20 Nonclassical Lymphocyte Subpopulations Objectives 1 In general identify how gd T cells NK cells and NKT differ from classic ab T cells in their antigen recognition 0 gd T cells amp NK cells amp NKT cells Recognize lipid antigens 0 More limited from limited receptor repertoires 0 Classic ab T cells Recognize pathogen peptides 0 Express a diverse array of ab TCR derived by the process of somatic recombination 2 Specifically identify the antigen that V99Vd2 T cells and VgVd1 T cells bind where these T cell populations are located in the body and what their general effector function is once activated V99Vd2 T cells 0 Present in adult human blood 0 Recognize and bind phosphoantigens small phosphorylated intermediates of isoprenoid biosynthesis O Majority cannot enter secondary lymphoid tissues so instead 80 located in inflamedinfected sites use CCR5 0 Function to secrete lFNy and TNFd and kill infected cells by granulysin secretion 0 Highly activated cells capable of antigen presentation VgVd1 T cells 0 Bind to MICA and MICB stress proteins 0 Located in blood of fetus and neonate but become confined to mucosa and spleen in adults 0 Function to kill damaged or infected gut epithelial cells leading to repair of injured mucosa 3 Identify the CD protein expressed on the surface of all human NK cells 0 CD56 and CD16 0 CD56 high NK cells 0 present in secondary lymphoid tissues 0 secrete large amounts of cytokines when activated 0 CD56 low NK cells 0 that circulate in the bloodstream looking for something to kill 0 More functionally mature and strongly cytotoxic 4 Compare and contrast MK and Tc target cell recognition and killing mechanisms Which of the following is true of NK cells 0 NK Cells 0 Innate immune cells 0 Viral immunity 0 Detect and kill virus infected cells by detecting the loss of MHC Class I on infected cells 0 Detect the balance of inhibitory and activating receptor signals to decide whether to kill or not 0 Tc Cells 0 Bind to polygenic self MHC l or MHC II To detect Pathogen peptides I Use CD4 or CD8 TCR Coreceptors O Creates both effector and memory cells 5 Describe the ligand for NK cell receptors in general and how the signals through these receptors are interpreted by the NK cell 0 NK cells target and kill virusinfected cells by detecting a loss of MHC Class 1 on the infected cell 0 The receptors fall into two categories lectinlike receptors chromosome 12 and lglike receptors chromosome 19 0 Both of these families contain inhibitory and activating receptors The ligands for these receptors are variable but often involve MHC Class 1 proteins 0 ligands are quite variable but often involve MHC1 proteins or class 1 like proteins in some way 0 lack of HLAE surface expression targets a body cell for killing 0 all NK cells express the activating receptor NKG2D kill cells expressing MlCA B Q CD94NKG2A is a common inhibitory NK receptor 6 Identify the specific ligand for the NKGZD receptor whether it is an activating or inhibitory receptor and what the NK cell will do if it encounters this ligand 0 The specific ligand for NKG2D is MICA and MICB Q NKG2D is an activating receptor and when an NK cell encounters this ligand the activating receptor NKG2D kills all cells expressing MICA or MICB I Describe how HLAE is involved in PREVENTING NK cell killing of healthy self cells 0 Lack of HLAE targets a body cell for killing 0 Surface expression of HLAE allows cells to escape the targeting 8 Identify what kind of antigen is presented by CD1 a CD1 b and CD1c molecules and what cell they present this unique antigen to Q CD1a through CD1c will bind foreign lipids that are synthesized by some pathogens but are not synthesized in human 0 They are presented to TCFIdB CD4 and CD8 T cells 9 Identify the classI like molecule that presents antigen to NKT cells where NKT cells are located in the body and what they do in response to antigen presentation that can influence a subsequent adaptive immune response 0 CD1d presents antigens to NKT cells 0 NKT cells are expressed by epithelial cells located in the liver intestine pancreas uterus thymus and tonsils Q NKT cells do not generate memory thus cannot influence an adaptive immune response 0 NKT cells respond to lipid antigen presented in CD1 D pg 24 Topic 21 Inherited Immunodeficiency amp Testing for Immunodeficiency Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the acronym SCID List the currently known mutations that can lead to SCID and the consequences of SCID in regards to infectious disease susceptibility ie is SCID associated with bacterial infections fungal infections viral infections or all of the above 0 SCID Severe Combined Immune Deficiency 0 Severe Combined Immune Deficiency Q Caused by absence of T and B Lymphocytes Q Xlinked SCIDS Caused by Common gamma chain cytokine receptor lL2Rgammac Q NonXlinked SCIDS caused by lack of Jak3 kinase 0 Or Adenosine Deaminase ADA Deficiency 0 Or loss of genes needed for somatic recomb RAGt or RAG2 example 0 Causes susceptibility to all infectious agents including simple fungi viral infections and bacterial infections 0 Or Purine nucleotide phosphorylase PNP deficiency 2 Describe the basis of DiGeorge Syndrome and the immune cell population that is defective as a result Explain why DiGeorge Syndrome can have variable levels of severity associated with it Q Digeorge Syndrome O Decreases of T lymphocytes because of Thymic Aplasia l Caused by loss of portion of Chromosome 22 O Caused by Lack of Tcell development in thymus l Affects people differently from mild to severe immunodeficiency 3 Identify what two primary immune deficiencies lead to selective susceptibility to viral infections and for each whether the patient is susceptible to all virus infections or just certain types of viruses 0 Bare Lymphocyte Syndrome 0 Type I MHC Class I Deficiency 0 TAP binding proteins don t allow for loading of peptide to MHC Class I I No Expression on surface 0 Lack of CD8 positive selection in thymus O SUSCEPTIBILITY TO ALL VIRUS NK Cell Absence O Susceptibility to Herpes virus infections 4 Explain why Type II Bare Lymphocyte Syndrome causes infectious disease susceptibility similar to SCID 0 Bare Lymphocyte Syndrome Type ll 0 Loss in transcriptional regulators of MHC Class ll 0 Causes lack of CD4 T cell positive selection in thymus O Resembles SCID and DiGeorge Syndrome 0 CD4 Tcell help for cell mediated and Humoral immunity is absent 0 CD8 intact but less functional less help from CD4 5 Compare and contrast Xlinked agammaglobulinemia and Xlinked HyperlgM syndrome specifically listing whether B cells are present in circulation and whether lgG lgA and lgM are absent or produced at reduced normal or excessive levels and the type of infection susceptibility associated with the diseases 0 Both cause susceptibility to extracellular bacteria esp encapsulated Q X linked agammaglobulinemia Absence of B lymphocytes due to Brutons tyrosine kinase Btk deficiency preB cells die in BM 0 Absence of all antibodies in the blood 0 All B lymphocytes gone no lgM production 0 Neisseria Sp susceptible Q Xlinked hyperlgM syndrome due to a lack of isotype switching because of CD40L deficiency neutropenia develops during infection because of coordinate loss of GM CSF by macrophages 0 B cells are responding and turning into plasma cells 0 Inability to have productive T cell help 0 Continue making lgM but never isotype switch to lgA or lgG Susceptible to pus forming bacteria pyogenic Most common in males 6 Identify why complement deficiencies are associated with increased susceptibility to extracellular pyogenic bacterial infections be sure to define pyogenic Q Pyogenic Causing the formation of pus Q Deficiency of C1 C2 or C4 early components of the classical pathway leads to immune complex disease can t activate well 0 Complement deficiency prevents immune complex clearance Coat immune complex with C38 0 susceptibility to encapsulated bacteria Complement deficiency resembles antibody deficiency in infection susceptibility Decreased opsonization thus decreased phagocytosis Q Deficiency of C59 increases susceptibility to Nesseria sp bacteria 7 Identify whether deficiencies of early components of the classical complement pathway the alternative complement pathway or the lectin complement pathway can cause a risk for systemic lupus autoimmunity and why this risk develops not in coursepack watch lecture 0 CR1 on RBC bind C3b Get C3b stuck on immune complex by classic pathway Q If missing early component of pathway 0 Immune complexes accumulate in blood and precipitate out in kidney 8 For the primary phagocyte deficiencies CGD ChediakHigashi syndrome G6PD or Myeloperoxidase deficiency and LAD identify the type of infectious disease susceptibility seen in patients and whether the infections respond to antibiotic treatment or not and why some do not respond to antibiotics 0 Chronic granulomatous disease 0 Patients lack a subunit of NADPH oxidase and can t activate granule proteases 0 Generally respond to antibiotics because inflammatory response is intact Q ChediakHigashi syndrome 0 Patients can t fuse endosomes and lysosomes 0 Generally respond to antibiotics because inflammatory response is intact Q G6PD or Myeloperoxidase deficiency 0 Patients have defective respiratory burst and impaired killing of phagocytosed bacteria and fungi 0 Generally respond to antibiotics because inflammatory response is intact 0 LAD Leukocyte adhesion deficiency 0 Patients have defective neutrophil and macrophage migration into inflamed tissues and inability to phagsocytize pathogens opsonized with iC3b O Infections respond poorly to antibiotics because inflammation becomes greatly diminished 0 chronic gum and mouth lesions 9 Identify the infectious disease susceptibility that results from defects in the lL12 or IFNg receptors and the immunological basis for this susceptibility Q Defects in these leads to combined defects in innate and adaptive immune func on o NK cell cooperation with macrophages is blunted because lL12 produced by macrophages cannot activate NK cell to produce IFN gamma or IFN gamma induced cannot feedback to activate macrophage production of inflammatory cytokines o lL12 is necessary for Th1 development and augments Tc production of IFNgamma o Leads to TB from mycobacterium Avium and active infection with mycobacterium bovis vaccine 10 List 2 routine laboratory tests that can be performed using patient blood samples serum or cells which are useful in evaluating possible primary immune defects in humoral immunity 0 lgG lgMlgA total antibody quantification serum analysis 0 Q B cell CD19 or 20 counts by flowcytometry Q Specialized testing for B lymphocyte defect o lgG Ab response to protein and polysaccharide Ag 0 In vitro B cell proliferation assay 0 Histology of lymph node biopsy 0 Serum protein electrophoresis to look for decrease or absence of gamma globulins only if the test is inclusive 11 Describe the ster of a Complement Activation Enzyme Immunoassay specifically identifying what is coated onto the assay EIA plastic plate wells the patient sample used the detection reagent used and how results are reported and what pathway of complement is being assayed Preplaces traditional complement hemolytic titration assay called the CH50 that measured the classical complement pathway components which was reported as the reciprocal of the dilution able to yse 50 of antibodysensitized sheep rbc CAE immunoassay uses EIA 96well plates coated with lgM and the detection reagent is enzyme linked antiC9 classical complement cascade must be triggered to completion and binding of C9 to the plate plastic Reported out as complement activity units Decreased activity indicates classical cascade complement factors are decreased usually due to consumption exceeding production while absent or zero activity units indicate a frank complement factor deficiency genetic mutation 12 Explain the flow cytometry technique including the reagents that are used the patient sample utilized and how that label is detected List what information can be collected on each cell analyzed and how that information is interpreted 0 Flow cytometry Used to assess specific cell populations in blood or any other fluid 0 A cell mixture is labeled with fluorescent tagged monoclonal antibodies specific for cell marker of interest 0 Cells are suspended in saline and run thru a flow cytometer I Separates cells into single cell stream I Activates the flurochrome chemicals that absorb light of one wavelength and emit light of higher wavelengths labels with laser light I Detects the fluorescent emission with detectors also can simply do forward cell size and side cell complexity laser light scatter Q Can be analyzed by a histogram or a two dimensional dot plot 0 Detects positive and negative cells 0 Amount of TCR on x axis 0 CD antigens useful for identifying lymphocyte subsets by flow cytometry o B cell CD19 or CD20 0 Total T cells CD3 0 T helper cells CD3 CD4 0 Tc cells CD3 CD8 o NK cells CD16 andor CD56 pg 26 Topic 22 HIVAIDS Virus and Disease Mechanisms Objectives At the completion of the modules and the assigned reading the student should be able to 1 Identify whether AIDS Acquired Immunodeficiency syndrome due to HIV infection is a primary or secondary immunodeficiency 0 AIDS is secondary Secondary immunodeficiency are people born with healthy immune systems that later become damaged acquired 2 Identify whether HIV1 or HIV2 is more common in the US 0 HIV1 3 State whether the number of people infected with HIV worldwide has leveled off declined or is still increasing 0 The number of people in the world with HIV is still increasing 4 Identify the function of gp120 and gp41 in the interaction with host cell membranes and eventual infection of host cells 0 GP120 on the HIV cell binds to CD4 and the co receptor on the TCell 0 Forms knobs protruding from envelope that bind CD4 amp CCR5 Allows virus to enter cell 0 GP41 adheres tighter and allows for membrane binding 0 The Viral envelope fuses with the cell membrane and the viral genome enters the cell allows virus to dump its content 5 Explain the difference between macrophagetropic and lymphocytetropic HIV strains and how they can change over time during HIV infection of an individual 0 Macrophagetropic HIV generally initiate infection at mucosal sites of entry by binding to CCR5 and normally infect macrophages and dendritic cells also CD4 T cells 0 Lymphocytetropic HIV strains preferentially bind CXCR5 and preferentially infect CD4 T lymphocytes later in infection 6 Explain the function and importance of reverse transcriptase and integrase HIV enzymes in creating a permanent stable infection of a host cell 0 When HIV infects a cell the RNA genome is first copied into a complementary DNA cDNA by reverse transcriptase The viral integrase then integrates the cDNA into the genome of the host cell to form a provirus a process facilitated by repetitive DNA sequences called long terminal repeats LTRs that flank all retroviral genomes Proviruses use the transcriptional and translational machinery of the host cell to make viral proteins and RNA genomes which assemble into new virions 7 Explain when the Acute HIV Syndrome occurs following exposure to HIV and identify the symptoms of this syndrome Is the infected individual infectious to others at this time Do all patients develop these symptoms 0 Western Blot assay for patients antibody against separated protein antigens 0 Acute HIV syndrome occurs within 48 weeks of contraction 0 Individuals are HIGHLY infectious Q There are Flu like symptoms to this phase I No all patients do not develop these symptoms Some of them are completely asymptomatic 8 Explain why CD4 T cell numbers temporarily plummet during the end of the acute infection stage of HIV and then rebound at least partially Q The number of CD4 T cells drops due to them being attacked by CD8 cytotoxic T cells There is then a slight rebound of CD4 levels due to stimulation of the bone marrowthymus realizing that CD4 levels have dropped significantly The rebound is also partially because the antibodies have slightly neutralized the virus allowing the T cells to come back 9 Define the acronym HAART and explain why HAART drug treatment is used in fighting HIV infection instead of single drug therapy 0 HAART Highly Active Antiretroviral Therapy 0 Use of HAART instead of just one drug 0 3 Different classes of Drugs allows each class to inhibit and help stop HIV with different mechanisms 0 HIV mutates extremely quickly 0 Use of 2 out of 3 drugs allows one drug to still work if the other fails 10 List the characteristics of HIV that make it very difficult to create an effective vaccine against HIV 0 HIV has a very high mutation rate 0 HIV can remain latent for long periods of time Q We can t identify cells that have it latent Q You have to target a Tcell response and Mucosal antibody response to be effective in stopping HIV infection pg 27 Topic 23 HIVAIDS Serology and Disease Monitoring Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms serology EIA window period false negative false positive assay sensitivity assay specificity indeterminate result 0 Serology Study of the noncellular components in the blood 0 EIA Enzyme immunoassay general type of test adaptable to many antigens 0 Window period Where someone could be infected but test negative false negative because test wasn39t sensitive enough 0 False negative Where a patient tests negative because the test wasn39t sensitive enough even if the patient was infected 0 False positive Give a result but really no antibody due to human cell contaminants Q Assay sensitive Must be sensitive enough to detect certain antigens Q Assay specificity Specific for certain antibodies 0 Indeterminate result One or more bands that do not meet the criteria for poste O Suggests that a patient should be resampled 2 Identify the two main analytes that can be measured in serological tests for infectious disease 0 Pathogen antigen 0 Host antibody 3 Interpret HIV screening and confirmatory test results using the CDC testing algorithm for HIV diagnosis For example if the screening test is negative the interpretation would be that the patient does not have HIV infection 0 Initial screening of serum from high risk individuals 0 if negative report as negative 0 if positive repeat in duplicate on same serum sample 0 if negative on both repeats report as negative Q If 2 of 3 tests from same serum sample are positive send for confirmatory testing confirmatory testing done with western blot 4 Correctly identify the next step to be taken in HIV diagnosis if the initial screening test is positive or if the repeat duplicate samples are positive 0 You do an EIA test and send for a confirmatory test Western Blot 5 Explain the basics of EIA testing for detection of either patient antibody or antigen in patient serum 0 Enzyme linked lmmunoAssay 0 General type of test 0 Adaptable to many antigens 0 Uses a solid support plastic wells nitrocellulose microbeads to detect antigenantibody reactions usually by addition of an enzymesubstrate pair that produces a colored product 0 Using EIA to detect patient Ab 0 Take solid Ag HIV viral antigens and patients antibody in diluted serum 0 Incubate wash and add next reagent enzyme antiglobulin conjugate wash again then it makes a substrate Q If there is no color its negative if colored positive 0 This type of immunoassay is easy to perform and high throughput many samples per plate many plates per run and is highly sensitive and specific 6 List the drawbacks of the early generation EIA tests for HIV diagnosis and how the fourth generation EIA test has limited these 0 First Generation 0 Ag was prepared from lysates of whole cultured human T cells infected with HIV1 O ProblemsBenefits l People could be infected but test negative for 45 days after the infection 0 Window Period I There were false positives because of contaminants I It could not detect HIV2 infections 0 Fourth Generation 0 Latest technology incorporates ultrasensitive detection of antibodies to HIV1 or HIV2 using recombinant proteins in a sandwich EIA format I ProblemsBenefits 0 More sensitive detection 0 The false negative time has dropped to 1421 days 0 Can detect both HIV1 and HIV2 Q There are still many false positives in low risk populations due to history of multiple pregnancies autoantibodies severe liver disease recent vaccinations certain malignancies repeated freezingthawing of specimen 7 Identify the confirmatory tests available for HIV diagnosis in adults assuming the screening test is positive 0 Western Blot General Test Principle 0 Manufacturer separates HIV proteins in gel blots transfers proteins to a membrane usually PVDF and cuts membrane into test strips 0 Laboratory scientist reacts membrane with patient serum as source of potential antiHIV antibody WASH O MLS adds enzyme conjugated antihuman globulin WASH O MLS adds substrate to detect patient antibody bound to individual HIV proteins each show up as a colored band on test strip 0 Assay for patients antibodv aoainst separated protein an gens 8 Explain how the western blot test adds specificity to the diagnosis of HIV infection and why we prefer a screening test to be more sensitive and less specific Q It interprets an Ab banding system you want a test to be more sensitive so it ll detect smaller amounts and be more accurate 0 Sensitivity number of time you get a positive test result in those who truly have the disease 0 False Negative were negative but really had the disease Greater the number of false negative the lower the sensitivity 0 of true positivesfalse negativestrue positives 0 Specificity people who test positive who truly have the disease 95 out of 100 have disease other 5 are false positive Specificity positive resultsfalse positive of positives 9 Identify the preferred methodology for detecting HIV infections in newborn infants and why the usual screening test for HIV can not be used to diagnose HIV infections in infants 0 Adult screening test can t be used to test baby s serum because of passive lgG antibody from the mother moms Ab in baby s serum can result in false positive test up to 18 months of age 0 Baby s lgM Ab to HIV lacks Specificity and is not adequate for testing 0 Best method is to test for viral antigen when baby is gt1 month old X2 0 Qualitative RTPCR for HIV cDNA provirus in baby s mononuclear cells 10 Correlate an increase or decrease in HIV viral load to the success of drug therapy in the patient Identify specifically what HIV viral load is measuring and the patient sample used 0 Successive drug therapy results in drop in viral load level 0 Drug resistance results in an increased viral load 0 The patient sample you use the measure the viral RNA is plasma because fluid portion 0 No proteins 0 Also virus small and soluble Q We are not detecting HIV proteins we are testing patient antibodies against HIV proteins 0 Generally use serum for these test l Serum is liquid after coagulation 11 Identify the laboratory test used in monitoring patient progression from HIV to AIDS 0 CD4 T cell count is 1 diagnosis for AIDS 0 Absolute CD4 T cell count lt200ul reference is 5001300ul 0 CD4 T cell of total lymphocytes lt14 0 CD4CD8 ratio lt11 reference 21 1 Define the following terms 0 Hepatitis general term for inflammation of the liver symptoms include fatigue fever GI upset muscle pain jaundice but many are asymptomatic following exposure and infection 0 Fecaloral route contaminated foodwater Q parenteral other than intestine means of needle stick sexual contact blood products etc Bloodbody fluids blood sexual contact etc HBcAg hepatitis B core antigen HBeAg hepatitis Be antigen HBsAg hepatitis B surface antigen antiHBc past infection indication appears later and persists for life antiHBs persists for years and provides protective immunity failure of a patient to develop these indicates chronic infection 2 Identify the main health concern associated with viral hepatitis acute chronic etc 0 The concern is usually chronic infections leading to possible cirrhosis andor liver cancer 3 Recognize the significance of lgM isotype antibody lgG isotype antibody and viral Ag or nucleic acid RNA or DNA in general staging of viral infections by laboratory testing 0 lgM and lgG are virusspecific antibodies in adult serum 0 lgM isotype antibodies indicates a current or recent acute infection 0 lgG isotype indicates a current or m infections and immunity 0 Virus specific lgM isotype in newborn serum indicates congential infections lgG does not cause could be from mom 0 Positive serological tests for viral AG or detection of viral DNA or RNA by molecular diagnostic testing indicates current infections 4 For HAV HEV HBV HCV and HBV identify the mode of transmission the incubation period whether the disease is only acute or becomes chronic what percent by age group develop chronic infection the longterm consequences of chronic disease and the antibodiesantigens that are useful for disease diagnosis and staging by serology testing It may be beneficial to create a study table with these column headings This table content will be most extensive for HBV testing and some content may not be applicable for all hepatitis viruses NOTE This is a very important learning objective Although it is only a single learning objective it usually results in 5 or more questions on the hourly examination HAV HBV HCV HDV HEV mode of fecaloral bloodbody bloodbody bloodbody fecaloral transmissio foodborne fluids fluids fluids n incubation 28 days 6090 days 78 weeks 37 weeks 38 weeks penod acutechron does NOT can become acute Can become does NOT ic can it become chronic symptoms chronic become become chronic but can chronic chronic become chronic by age most infected 510 of 85 All age developing group that adults are SX infected groups countries develop children adults pregnant chronic usually aSX 3060 of women infection infected children 90 of infected infants longterm NA 1 million develop liver Can lead to WA consequen deaths per cirrhosis can cirrhosis and ce of year develop hepatocellula chronic worldwide heptocellular r carcinoma infection circinoma AbAgs that lgM antiHAV HBcAg antiHCV lClM antiHDV loM anti are useful indicates HBeAg detected by appears 67 for disease acute HAV indicates high EIA and Ab weeks after serological Dx high total virus appears 7 to exposure test antiHAV Ab infectivity 8 weeks later lgG antiHDV indicates HBsAg and has 99 appears immunity peaks during specificity during acute false convalescenc infection positive e then lClM antiHBC HCV RNA declines after indicator of molecular resolution fo acute assay for the infection infection patient s lgM antiHDV lgG antiHBC serum this is and lgG anti indicates confirmatory remain past infection testing Use high in AntiHBs RIBA chronic failure to infection develop gt chronic infection 5 Explain how a RIBA test such as that used to confirm HCV infection differs from a western blot test Q It is like the Western Blot except Recombinant HCV proteins are blotted directly onto nitrocellulose membrane 6 Given the results of hepatitis panel tests for acute viral hepatitis interpret the disease that the patient has and suggest whether further testing is warranted and what that testing should be ordered 0 When a patient presents with jaundice or other signs of acute hepatitis usually a hepatitis panel is ordered 0 Hepatitis A lgM antiHAV O Hepatitis B lgM antiHBc and HBsAg can run one and then run the other for confirmation 0 Hepatitis C AntiHCV and HCV RNA 0 Panel results help determine whether hepatitis is due to a viral infection or not and which virus is causing the infection HBcAg Core antigen HBeAg Be antigen HbsAg surface antigen 1 st to appear in serum Topic 25 Herpes Viruses EBV CMV HSV and VZV amp Rubella 1 Define the following terms including what cells the term applies to Q TORCH testing Toxoplasmosis other infections Rubella Cytomegalovirus and Herpes Simplex Virus is often performed in young women of childbearing age bc of consequences of acute infection in utero or for newborns if acute infection is transmitted from mom to baby 0 titer Concentration of virus 0 heterophile antibody lgM antibodies that are capable of reacting with similar antigens from two or more unrelated species 0 herd immunity protection from spread of infection by vaccinationinduced population immunity 2 Identify which viruses other common names belong to the Herpes Virus Family 0 EpsteinBarr virus EBV or HHV4 Q Cytomegalovirus CMV or HHV5 Q Herpes Simplex viruses HSV1 and HSV2 Q VaricellaZoster virus VZV or HHV3 0 other human herpes viruses HHV6 HHV7 HHV8 3 Identify the disease caused by EpsteinBarr Virus EBV the immune cell target of EBV and the receptor that it uses to target that cell and the effect of EBV on the infected cell 0 causes infectious mononucleosis Q infects epithelial cells of the oropharynx and B lymphocytes via CD21 0 infected B cells undergo polyclonal activation proliferate and secrete antibody 4 Associate detectionscreening for heterophile antibodies with diagnosis of infectious mononucleosis Q heteroohile antibodies are lgM antibodies that are capable of reacting w similar Ag from 2 or more unrelated species 0 heterophile Ab are found in other cases besides IM but the term heterophile Ab is now usually reserved to designate IM 0 for IM screening w Monospot test heterophile antibodies are detected w extract of bovine cow RBC Ags 5 Identify confirmatory testing for EBV infection and the significance of these tests in disease staging 0 about 20 of pts w EBV don t make heterophile Ab and give false negative testing in the Monospot must differentiate when st could also be associated w CMV Toxoplasmosis or Adenovirus infections 0 IClM Ab to viral cabsid ACl lgM antiVCA if indicates acute disease and shows up early in infection 0 lgG antiVCA also present during late acute IM but persists and could indicate previous infection Q Antibodv to EBV nuclear antigen antiEBNA is the last pt antibody to appear showing up during convalescence and persists for life 6 Identify the patient populations at greatest risk from cytomegalovirus CMV infection and the possible consequences of infection ADD stuff on bottom of 183 High percentage of adults 90 usually asymptomatic in healthy individuals but can be very severe in immunocompromised and newborns CMV persists in latent state in myeloid cells Most common cause of congenital infections throughout the world 10 percent of infected babies exhibit symptoms of CNS andor multiple infection 0 5 of these infants die 0 50 of surviving symptomatic babies have longterm consequences such as hearing loss mental retardation vision loss 7 Identify the problems associated with serological testing for antibody to CMV and the preferred methods of detecting CMV infection in the atrisk populations 0 serological tests for CMV must be interpreted carefully O assays for lgM antiCMV have false negatives in newborns and immunocompromised and false positives from rheumatoid factor RF 0 rapid detection not readily available need to document 4fold rise in lgG antiCMV titer in two serial specimens drawn 4 weeks apart 0 newest EIA assay for acute disease diagnosis measures low avidity lgG antibody prior to affinity maturation not widely available in US yet 0 the most reliable diagnostic tests for acute CMV infection are performed on saliva urine or throat swab samples and include 0 shell vial cultures for viable virus can detect infection in 2472 hrs specificity from immunofluorescent staining w monoclonal antibodies 0 qPCR for CMV DNA also gives rapid results and can help to stage disease and monitor effectiveness of antiviral therapy 8 Identify the target tissue of HSV1 and HSV2 viruses and which is of most concern 0 HSV1 is cause of oral herpes Q HSV2 is cause of herpes genitalis genital epithelium most serious of the two HSV infections 9 Identify the testing available for detection of HSV1 and HSV2 infection whether the Ag or Ab to the virus is detected by serology and which test is currently able to distinguish between HSV1 and HSV2 Q Serological testing by EIA detects viral Ag 0 PCR test detects viral DNA and can distinguish between HSV1 and HSV2 10 Identify the two diseases caused by VaricellaZoster Virus VZV and the reason why the viral infection reactivates Q Varicella aka Chicken Pox as an acute disease 0 Herpes zoster aka Shingles as a reactivation of latent disease 0 Following resolution of primary chicken pox VZV retreats to sensory nerve dorsal ganglion cells and reactivates in about 20 of individualsunder stress as Shingles Dont know why exactly it reactivates it s stress induce reactivate virus when stressed 11 Identify the methods used to detect VZV infection versus immunity to VZV 0 Shell vial culture method is coupled with immunofluorescent staining to identify the virus PCR detection however is rapidly becoming the test of choice 0 Serology EIA is used for establishing immunity to VZV over 90 of the population is immune either from natural infection or vaccination by live attenuated varicella vaccine 4 fold rise in paired acuteconvalescent titers is diagnostic 12 Identify the disease caused by Rubella virus and the most serious infection caused by Rubella 0 caused by German Measles Q contagious and acquired from droplets nasopharyngeal secretions 13 Identify the methods used to detect Rubella infection and how a significant titer is interpreted in regards to postvaccination or as an indication of current infection 0 Rubella serology 0 post vaccination antirubella lgG antibody level gt15 lUmL indicates immunity in prospective mothers 0 EIA with synthetic rubella peptides as the test antigen is the most common serology test 0 Pregnant mothers with suspected acute infection a 4fold rise in lgM titer in sample drawn 5 days apart indicate current acute infection 0 lgM antiRubella in newborns indicate congenital disease confirmed by positive culture or PCR 0 NO lgM or lgG means person not vaccinated pg 28 Topic 24 Hepatitis Viruses Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms Q Hepatitis General term that means inflammation of the liver Can be grouped by mode of transmission 0 Fecaloral route Contaminated foodwater Q Parenteral Bloodbody fluids 0 Bloodbody fluids Parenteral Q HBcAg O Hepatitis B core antigen a protein surrounding viral DNA in virus core 0 Not detectable in serum 0 HBeAg O Hepatitis Be antigen also a protein surrounding viral DNA in virus core 0 Appears shortly after HBsAg and indicates high viral replication and a high degree of infectivity Q HBsAg O Hepatitis B surface antigen a protein in outer virus envelope and in particles in blood O The first HBV antigen to appear in serum l Detectable 212 weeks after exposure I Peaks during acute infection I Levels decline as patient Ab increases usually undetectable 1220 weeks after symptoms l Persistent HBsAg is an indication of chronic or active infection I Not detectable in serum 0 AntiHBs 0 Appears during recovery 0 Persists for years and provides protective immunity 0 Failure of infected patient to develop antiHBs indicates chronic infection 0 This antibody is produced in response to vaccination Q AntiHBc O lgM Appears first I Used as an indicator of current or recent acute infection I Useful for detecting core window time between disappearance of HBsAg and detectable antiHBs O lgG Appear later and persists for life I Indicates past infection 2 Identify the main health concern associated with viral hepatitis acute chronic etc Associated with high rate of mortality in pregnant women who become infected 3 Recognize the significance of lgM isotype antibody lgG isotype antibody and viral Ag or nucleic acid RNA or DNA in general staging of viral infections by laboratory testing lgM Indicates a current or recent acute infection lgG Indicates current or past infection and immunity in most cases 4 For HAV HEV HBV HCV and HDV identify the mode of transmission the incubation period whether the disease is only acute or becomes chronic what percent by age group develop chronic infection the longterm consequences of chronic disease and the antibodiesantigens that are useful for disease diagnosis and staging by serology testing It may be beneficial to create a study table with these column headings This table content will be most extensive for HBV testing and some content may not be applicable for all hepatitis viruses NOTE This is a very important learning objective Although it is only a single learning objective it usually results in 5 or more questions on the hourly examination HAV 5 Explain how a RIBA test such as that used to confirm HCV infection differs from a western blot test 6 Given the results of hepatitis panel tests for acute viral hepatitis interpret the disease that the patient has and suggest whether further testing is warranted and what that testing should be pg 29 Topic 25 Herpes Viruses EBV CMV HSV and VZV amp Rubella Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms including what cells the term applies to TORCH testing titer heterophile antibody and herd immunity ToRCH testino Toxoplasmosis other infections Rubella Cytomegalovirus and Herpes Simplex Virus 0 Often performed in young women of childbearing age because of consequences of acute infection in utero or for newborns if acute infection is transmitted from mother to baby m Reciprocal of last dilution that yields a positive test in assay Heteroohile Antibodv O lgM antibodies that are capable of reacting with similar antigens from two or more unrelated species for EBV sheep and cows are the unrelated species 0 Heterophile Ab are found in other cases besides IM but the term heterophile antibody is now usually reserved to designate IM 0 For IM screening with Monospot test heterophile antibodies are detected with extract of bovine cow rbc antigens Herd lmmunitv Prevent pregnant women from being exposed to rubella 0 Protection from the spread of infection by vaccinationinduced population immunity 0 95 have to be vaccinated to have herd immunity 2 Identify which viruses other common names belong to the Herpes Virus Family EpsteinBarr Virus 0 Also known as EBV or Human Herpes Virus4 HHV4 Cytomegalovirus O CMV O HHV5 Herpes Simplex Viruses O HSVt O HSV2 VaricellaZoster Virus 0 VZV O HHV3 0 Other Human Herpes Viruses O HHV6 O HHV7 O HHV8 3 Identify the disease caused by EpsteinBarr Virus EBV the immune cell target of EBV and the receptor that it uses to target that cell and the effect of EBV on the infected cell 0 CD56 and CD16 0 There is CD56 high NK cells that secrete large amounts of cytokines when activated and CD56 low NK cells that circulate in the bloodstream looking for something to kill 4 Associate detectionscreening for heterophile antibodies with diagnosis of infectious mononucleosis 0 Cause of Infectious Mononucleosis IM aka Mono in young adults asymptomatic in children 0 Fever 0 Lymphadenopathy O Sore Throat O Lethargy Q EBV infects epithelial cells of the oropharynx and B lymphocytes via CD21 0 Infected B cells undergo polycoona activation proliferate and secrete an body O EBVspecific antibody 0 Autoantibody O Heterophile antibody used for diagnosis I Heterophile antibodies are lgM antibodies that are capable of reacting with similar antigens from two or more unrelated species for EBV sheep and cows are the unrelated species I Heterophile Ab are found in other cases besides IM but the term heterophile antibody is now usually reserved to designate IM I For IM screening with Monospot test heterophile antibodies are detected with extract of bovine cow rbc an gens O Other EBV Serology l About 20 of patients with EBV do not make heterophile Ab and give false negative testing in the Monospot must differentiate when symptoms could also be associated with CMV Toxoplasmosis or Adenovirus infections 5 Identify confirmatory testing for EBV infection and the significance of these tests in disease staging 0 K cells target and kill virusinfected cells by detecting a loss of MHC Class 1 on the infected cell The receptors fall into two categories lectinlike receptors and lglike receptors Both of these families contain inhibitory and activating receptors The ligands for these receptors are variable but often involve MHC Class 1 proteins 6 Associate detectionscreening for heterophile antibodies with diagnosis of infectious mononucleosis Q Heterophile antibodyused for diagnosis of EBV mono lgM Ab that are capable of reacting with similar antigens from two or more unrelated species 0 Found in other cases besides mono but the term is now reserved to designate infectious mono Q Detected with extract of bovine cow RBC Ag 7 Identify confirmatory testing for EBV infection and the significance of these tests in disease staging 0 About 20 of patients with EBV do not make heterophile Ab 0 False negatives Q lgM Ab to viral capsid Ag lgM anti VCA indicates acute disease 0 lgG antiVCA also present during acute IM but persists and could indicate previous infection 0 Antibody to EBV nuclear antigen antiEBNA appears during convalescence when you start to feel better 8 Identify the patient population at greatest risk from cytomegalovirus CMV infection and the possible consequences of infection Q CMV is the most common cause of congenital infections throughout world 032 of all live births Q 10 of infected babies exhibit symptoms of CNS and or multiple organ infection 0 5 of these infants die 0 50 of surviving symptomatic babies have long term consequences such as l Hearing loss I Mental retardation l Vision loss 9 Identify the problems associated with serological testing for antibody to CMV and the preferred methods of detecting CMV infection in the atrisk populations Rapid detection not available need to document 4 fold rise in titer in two serial specimen drawn 4 weeks apart Assays for lgM antiCMV have false negatives in newborns and immunocompromised and false positives from rheumatoid factor RF Shell vial cultures for viable virus can detect infection in 2472 hours specificity from immunofluorescent staining with monoclonal antibodies Quantitative PCR for CMV DNA also give rapid and can help to stage disease and monitor effectiveness of antiviral therapy 10 Identify the two diseases caused by VaricellaZoster Virus VZV and the reason why the viral infection reactivates Varicella chicken pox an acute disease Herpes zoster shingles reactivation of latent disease Following resolution of primary chicken pox VZV retreats to sensory nerve dorsal ganglion cells and reactivates in 20 o of individuals under stress as Shingles Shingles is eruption of a painful blistering rash over the site of the nerve where VZV was latent Usually occurs in those who have decreased Tc cellmediated immunity Particularly severe in AIDS patients and the elderly 11 Identify the methods used to detect VZV infection versus immunity to VZV Shell vial culture method couples with immunofluorescent staining to identify the virus PCR for VZV DNA detection is rapidly become the lab test of choice Serology ELISA used for establishing immunity to VZV gt90 o of population is immune either from natural infection or vaccination by live attenuated varicella vaccine 12 Identify the disease caused by Rubella virus and the most serious infection caused by Rubella Rubella is a RNA virus that is not related to herpes family Causative agent of german measles which may be acquired from infected people or congenitally wasn t seen in US because of MMR vaccines Acquired rubella infection german measles Rubella is very contagious and acquired from droplets of nasopharyngeal secretions Mostly school ages children get infected Symptoms are usually benign rash low fever headache Natural disease of vaccination provides lasting immunity Congenital rubella infection RARE but bad Fetus may be stillborn Severe bone or cardiovascular defects Mental retardation Microencephaly Hepatomegalysplenomegaly Cataracts High incidence of IDDM later in life Vaccine should be given if pregnant Vaccine program exists to herd immunity and prevent bad infection 13 Identify the methods used to detect Rubella infection and how a significant titer is interpreted in regards to postvaccination or as an indication of current infection Post vaccination gt1 8 titer of lgG suggests immunity 1015 lUmL Pregnant mother with suspected acute infection a 4 fold rise in lgM titer in samples drawn 5 days apart indicates current infection Eg 14 titer on first sample 116 titer in second sample would be indicative of mother with acute infection when samples are tested at the same time by the same lab lgM antirubella in newborn indicates congenital disease confirmed by positive culture or RTPCR for rubella RNA No lgM or lgG means the person has not been vaccinated or infected EIA with synthetic rubella peptides as the target test antigen is most common serological test TITER highest dilution of patient solution that still gives positive result Topic 26 Spirochete Serology Syphilis amp Lyme Disease Objectives At the completion of the modules and the assigned reading the student should be able to a Get it from Treponema pallidum ssp pallidum spirochete b Transmitted from sex or mother congenital c Treat it with penicillin in early stages 2 Define the following terms nontreponemal tests treponemal tests reagin Q Nontreponemal tests Use a test antigen consisting of a solution of cholesterol lecithin and cardiolipin to detect reagin antibodies 0 Ag it uses is artificial Q Treponemal tests Detect antibody specifically to treponemal antigens Q Reagin Non specific antibodies to cardiolipin causes positive result in nontreponemal tests for syphilis Q Treponemal Ab against the organism itself 0 Nontreponemal Ab against component that show up in cell damage 3 List the 4 stages of syphilis the symptoms of each stage and the timeframe in which each stage may be observed following initial infection 0 Primary syphilis Well defined painless lesion chancre with an ulcerated center and red raised border forms 16 weeks 0 Usually on genitals O Lesion will resolve on own if untreated 0 Secondary syphilis O Occurs in 25 of untreated infections 1 2 months after primary 0 Organism spreads through body and causes generalized lymphadenopathy and flulike O lesions common on trunk palms of hands and soles of feet are highly contaigous Q Latent syphilis O No symptoms for many years despite organism presence lasts 1030 years 0 Tertiary syphilis O Develops 1030 years later in about 30 of those who have latent syphilis Shows up in brain heart muscle and bone symptoms 4 List the three general types of testing available for diagnosing syphilis the usual use of each general type of test and categorize each specific test discussed in lecture into each of these general types 0 Direct spirochete detection in lesion fluid based on corkscrew shape and flexing motility not specific for syphilis because of normal flora Q Nontreponemal testsuse a test antigen consisting of a solution of cholesterol lecithin and cardiolipin to detect reagin antibodies 0 Two classic screening tests use this method I VDRL venereal disease research laboratory of CDC 0 Cholesterol lecithincardiolipin suspension mixed with patient serum forms flocculation microscopic precipitation in positive test 0 Not common and labor intensive and need a fresh sample daily 0 This is the ONLY syphilis test approved to test CSF to diagnose neurosyphilis l RPR rapid plasma reagin test 0 Patient plasma can be used as sample instead of serum and does not need to be heat inactivated to remove complement activity 0 Same basic reagent as VDRL except added charcoal particles allow macroscopic flocculation l Nontreponemal ElA s also exist that coat 96 well plates with cholesterol and lecithin and cardiolipin l Non treponemal tests detect lgG Ab which can be from mother so these tests can t be used to diagnose congenital syphms Q Treponemal testsdetect antibody specifically to treponemal Ag 0 EIA an ClAs treponemal lysate or recombinant treponemal proteins detect either lgM or lgG Ab to syphilis using antibodycapture technique most EIACIA testing currently this type 0 FTAABS fluorescent treponemal Antibody Absportion Tests 0 traditional confimatory test for syphilis O T pallidum organisms are fixed on a slide patient serum is added bound patient antibody is detected with fluorescent labeled antihuman globulin Q TPPA T pallidum particle Agglutination 0 Gel particles coated with treponemal lysate Ag or recombinant treponemal Ag 0 Positive test gel particles agglutinate in presence of patient serum 0 Syphilis tests vary in sensitivity during stages of disease 5 List the components of the test antigen reagent used for nontreponemal tests for sypth 0 Cholesterol lecithin cardiolipin for VDRL 0 Additional charcoal for RPR a VDRL Venereal Disease Research Laboratory of the CDC i Suspension of cholesterol lecithin cardiolipin from beef heart is mixed with patient serum flocculation microscopic precipitation is a positive test resuH ii This is the ONLY syphilis test approved to test CSF to diagnose neurosyphilis serum VDRL is not sensitive iii A positive test result is tittered by patient sample dilution b RPR Rapid Plasma Reagin Test i Patient plasma can be used as a sample instead of a serum hence the name and does not need to be heatinactivated to remove C activity ii Same basic reagent as VDRL except added charcoal particles allow macroscopic flocculation iii A positive test result is tittered by patient sample dilution Nontreponemal ElA s also exist that coat 96 well plates with cholesterol lecithin cardiolipin Example Reagin ll Assay 6 Identify the only test that is currently approved to diagnose tertiary neurosyphilis and the patient sample used for this testing 0 Nontreponemal test VDRL is used for CSF testing cerebral spinal fluid instead of serum for neurosyphilis 7 Identify which type of test for syphilis loses sensitivity during the tertiary phase of infection ie can give a false negative result in a patient who really has tertiary sypth Q VDRL and RPR nontreponemal tests Q List the proper order of screening and confirmatory testing for syphilis as it was classically performed and how this has changed now with newer reversed order syphilis serology testing a The traditional order was with nontrep As screening test and rep as confirmatory test Now trep EIA or CIA in 96 well plate is used for screening and nontrep RPR is used to confirm 8 Identify the causative agent of Lyme disease the characteristic symptom of primary infection that is observed in 80 of patients and disease sequelae that may be seen in chronic infection 0 Caused by spirochetes O Organisms causing infection are difficult to culture 0 Infections can lead to chronic sequelae resulting from in untreated patients 0 Tick borne infection caused by Borrelia burgdorferi Q Often a rash forms around the site of infection Erythema chronicum migrans rash bullseye rash is seen by about 80 of patients in about one week following bite Q If untreated organism spreads through bloodstream and may cause multiple lesions at sites distant from the bite Q Migratory pain in joints tendons muscles and bones may be exhibited 0 About 15 of patients have neurologic or cardiac involvement about 6 weeks after onset of infection 0 Most common chronic symptom is large joint arthritis 0 Facial palsy may be seen as neurological symptom 0 Enzyme immunoassays EIA or lmmunofluorescent assay IFA are traditional screening tests run with patient serum samplesllack specificity and sensitivity 0 Patient samples with equivocal or positive screening test are retested using western blot confirmatory test 0 New C6 B burgdorferi lyme ELISA has 99 specificity and has sensitivity 10 Identify possible causes of biological false positive results in Lyme disease serology testing Syphilis and other treponemal diseases Infectious mononucleosis EBV Autoimmune diseases Rocky mountain spotted fever pg 31 Topic 27 Group A Streptococcus Objectives At the completion of the modules and the assigned reading the student should be able to 1 List the main distinguishing features of Group A B hemolytic streptococcus S pyogenes used to identify the organism in a microbiology laboratory 0 Grampositive cocci that grows in chains 0 Belongs to serotype Lancet group A 0 based upon surface M protein 50 serotypes total 2 Define M protein and SpeA1 and identify which is a super antigen 0 M Protein Helps the organism evade innate immune cell defenses Q Streptococcal pyrogenic exotoxin A1 SpeA1 is a second virulence factor that acts as a super antigen 3 Describe the characteristics of these virulence factors that help S pyogenes escape the host immune response 0 Two major virulence factors and MANY minor virulence factors 0 M protein helps the organism evade innate immune cell defenses l Prevents phagocytosis l Inhibits C3b deposition on bacterial surface 0 Streptococcal pyrogenic exotoxin A1 SpeA1 is a second virulence factor that acts as a superantigen 4 Describe how a super antigen activates T lymphocytes 0 Causes NonAntigen Specific activation of T Lymphocytes O Physically holds the MHC thing and TCR together 0 This causes the T cell to be activated even if it is not specific for that an gen Q Can cause Cytokine storm large release of cytokines if 1045 of T cells activated 5 Describe the main clinical features of Impetigo and Group A Streptococcal pharyngitis Identify which or both is associated with Scarlet Fever Rheumatic Fever Acute Proliferative Glomerulonephritis and Toxic Shock Syndrome 0 Impetigo skin infection that usually occurs in young children on the face 0 Associated with l Acute proliferative glomerulonephritis destruction of kidney glomerulus by deposition of exotoxin l Toxic shock syndrome TSS Cytokine Storm from massive T cell activation leads to peeling skin shock due to lack of volume control etc 0 Group A Streptococcus 0 Secondary sequelae can result from primary infection apparently from antibody response to the organism l Scarlet fever Strep exotoxin itself creates characteristic body rashpeeling skin associated with Strep throat rarely seen in US due to antibiotics l Rheumatic fever Joint pain carditis murmurs due to crossreactive antibodies binding to cardiac myosin arterial smooth muscle cells and muscle glycogen 23 weeks following strep throat 1 risk following untreated Strep pharyngitis l Acute Proliferative Glomerulonebhritis Destruction of kidney glomerulus by deposition of exotoxin Q Antibody immune complexes and C lysis following either strep throat or streptococcal pyoderma most common 0 Toxic Shock Syndrome TSS 0 Due to superantigen of S pyogenes and also associated with superantigen of S aureus 0 Cytokine Storm from massive T cell activation leads to peeling skin shock due to lack of volume control etc 6 What is the main drawback of rapid testing for Strep Throat Q Strep throat easily diagnosed by rapid immunochromatography or latex antigen test 80 o sensitive O Negative rapid strep testing should be followed by culture in cases of suspected Strep pharyngitis 0 Major drawback is that is lacks sensitivity 7 Compare and contrast the efficacy of the ASO antiDNase B and Streptozyme assays in diagnosis of past Group A Strep infections Impetigo and Strep Throat 0 A80 antistreptolysin O test 0 Historical red blood cell lysis assay rarely performed in clinical labs any longer 0 Newer version of assay is performed by nephelometry to detect streptolysin O reagentantibody patient immune complexes Q Is positive in 80 of strep throat cases but does NOT become positive in Strep pyoderma aka it LACKS sensitivity O The A80 antibodies are produced a week to a month after initial step infection 0 AntiDNase B O AntiDNase B assay is more sensitive than A80 and will be positive in cases of streptococcal pyoderma I Can be performed by colorimetric assay anti DNase B blocks reagent DNase enzyme activity that would otherwise clear a green solution of DNA EIA or nephelometry l Nephelometry is the most cost effective 0 Streptozyme O Streptozyme test is a slide red blood cell agglutination screening test I Sheep rbc are coated with streptolysin streptokinase hyaluronidase DNase and NADase and mixed with 1100 dilution of patient serum l Significant false positives and false negatives 0 Recommended that at least two separate test formats be performed to achieve 95 sensitivity for Group A strep sequelae detection FINAL EXAM INFORMATION 0 Half old 0 Material that is mentioned over and over again 0 Differences between adaptive and innate immunity 0 How are B cells formed versus how T cells formed I How they function I What are the differences 0 Heavily based off Exam 1 material I Except not all exam 1 so keep that in mind I Focus on other main concepts in other exams 0 Half new pg 32 Topic 28 Hypersensitivity Responses Objectives Topic 28 Hypersensitivity Responses Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms RIST RAST Q RIST Radioimmunosorbent Test Total serum lgE measured by chemiluminescence as a screening tool REGARDLESS OF Ag SPECIFICITY ADDITIONAI INFORMATION Q Radioimmunosorbent Test a competition test performed in Vitro used to measure IgE specific for a particular antigen Known amounts of radiolabeled IgE compete with the patient39s unlabeled IgE to bind to a surface coated with antilgE The reduction in radiolabeled IgE due to the presence of IgE in the patient39s serum can be determined by comparison with known IgE standards thus the amount of the patient39s total serum IgE can be determined 0 RAST Radioallergosorbent Test Laboratory testing for allergenspecific lgE Current technology can still be called RAST even though radioactivity is not used in the assay Specific to a particular ag ADDITIONAL INFORMATION 0 Radioallergosorbent Test a radioimmunoassay test to detect specific IgE antibodies responsible for hypersensitivity the allergen is bound to insoluble material and the patient39s serum is reacted with this conjugate if the serum contains antibody to the allergen it will be complexed to the allergen Radiolabeled antihuman IgE antibody is added where it reacts with the bound IgE The amount of radioactivity is proportional to the serum IgEAd 2 Explain the immune system mechanism responsible for each of the 4 types of hypersensitivity reactions including the isotype of the antibody involved as appropriate andor effector cell types acting in each Type Features Conditions Type I IgE mediated Asthma mast cell degranulation Hay Fever Type II Abmediated cell surface rxns Hemolytic anemia that cause cytotoxicity HDFN complement activation Type III Immune complex mediated Arthus rxn complement activation Type IV cell mediated Contact dermatitis sensitized T cells activated macrophages 3 Explain how an immediate hypersensitivity differs from a delayedtype hypersensitivity DTH response and what type of hypersensitivity I IV is being tested for by each 0 DTH type 4 hypersensitivity 0 Develop symtpoms in 13 days following contact with allergen O A TB skin test is an example of a DTH response testing for cellular immune response to mycobacterium tuberculosis 0 Read 48 hours later unlike allergy skin testing which is read in 30 minutes 0 Wheal and flare skin testing greater or equal to 3mm swelling in 15 minute 0 Only tests limited individual allergens O Type1 hypersensitivity O Mast cells degranulate 0 Immediate and late phase responses right away and 6 hours later 4 Identify which hypersensitivity is associated with histamine release 0 Type I hypersensitivity O Histamine is 10 of total mast cell granule contents helps overcome the allergen responses Q It increases vascular permeability and causes smooth muscle contraction 5 Identify the major granulocyte that plays a role in allergic and parasitic diseases the antibody isotype that triggers its degranulation and the unique components of this cells granules Q Mast cell 0 Antibody lgE triggers its degranulation O Granules both cause inflammation l 10 percent Histimine l Histamine PG Leukotrienes 6 Describe the utility of total serum lgE testing skin prick testing and allergen specific laboratory testing listing the advantages and disadvantages of each Total serum lgE testing 0 An elevated total serum lgE Suggests type 1 hypersensitivity 0 Advantages l Cheap l Suggests further testing 0 Disadvantages I Not sensitive 13 ppl with allergies have normal lgE l Does NOT identify the offending allergen Skin allergy testing 0 Cutaneous or intradermal performed by allergist or nurse 0 Graded for wheal reaction gt 3mm swelling in 15 min 0 Advantages l A positive test is clinically significant 0 Disadvantages l Danger of systemic reaction I Traumatic I Only tests certain allergens Allergen specific allergen laboratory testing 0 Historical ways of testing 0 Radioimmunosorbent RIST and radioallergosorbent RAST l Traditionally immunoassays utilizing radiolabel to detect low levels of circulating human lgE I Was set up to determine total lgE or allergen specific lgE I No longer used bc current testing is done with enzyme fluorescence or chemiluminescence 0 Current technology I Testing is done on large automated analyzers single EIA Q Can test total lgE or a huge array of individual common allergens 0 Solid phase carbohydratebased assay agarose beads crystalline cellulose particles cellulose disks used to provide high test antigen dose for increased sensitivity l Lower specificity than skin testing 0 Only tells if lgE is present NOT if responsible for current patient symptoms I Can be used when patients are taking antihistamine l Performed in large panels 0 Lab testing for lgE microarray panel testing I Advantage is that it greatly increases the number of allergens that can be tested per sample 0 Glass platform based printed microarrays O Purified allergen is dotted on a glass slide in a predetermined pattern 0 Patient sera is added 0 AntilgE fluorochrome is added and the fluorescent dots are read via automation Q Bead array 0 A mixture of beads of different sizes and containing built in identifying fluorochrome are coated with individual allergens mixed with patient serum labeled with detection antilgE fluorochrome and run thru a flow cytometer 7 Identify common causes of type II hypersensitivities Hemolytic anemia Thrombocytopenia decreased platelets Also responsible for blood transfusion reactions and some autoimmune diseases Drugs bind to cell surface creating novel antigen B Cell respond and produce lgG antibody Upon 2nd exposure preformed lgG coats cells can cause direct C lysis or uptake by phagocytosis inflammation induced by phagocyte activation 8 For the following illnessesdiseases identify the type of hypersensitivity responsible serum sickness celiac disease poison ivy hypersensitivity allergic asthma food allergy such as shellfish allergy serum sickness type 3 celiac disease type 4 DTH poison ivy hypersensitivity type 4 DTH allergic asthma type 1 food allergy such as shellfish allergytype 1 9 Identify which hypersensitivity responses involve complement activation 0 Type III hypersensitivity is due to immune complex deposition activating complement which leads to in ammation Type 2 has direct complement lysis or uptake by phagocytosis Topic 29 Autoimmunity Organspecific Diseases Objectives At the completion of the modules and the assigned reading the student should be able to 1 Identify which types of hypersensitivity responses can cause autoimmune disorders and which of these is most commonly encountered 0 Type 2 Autoantibodies against cell surface or extracellular matrix proteins MOST COMMON 0 Type 3 Formation of soluble immune complexes that deposit in tissues joints or kidney glomeruli esp leads to systemic autoimmunity 0 Type 4 Effector t cells directly destroy body cells 0 lgE Type 1 is NEVER responsible for autoimmune diseases 2 Be able to match the autoimmune endocrine disorders Chronic Autoimmune Thyroiditis Grave s disease Addison s disease and Type 1 diabetes mellitus with the autoantibodies that may be observed in each disease the serological testing done to diagnose the disease if appropriate and the immune mechanism that is responsible for the observed tissue destruction 0 Grave s diseasehyperthyroidism 0 Due to Ab to thyroidstimulating hormone receptor anti TSHR Llil39 w T 39 cam men autoimmune 0f I aritfhiaut nee for r m Tl If i glam Nth0 h ute i it u n test ruc n thyri epithelial I viiag mloulin pie I quotl P nt in of h atients but I 0 Both can be assayed by enzyme immunoassay particle agglutination or indirect immunofluorescence fixed sections of monkey thyroid tissue incubated with patient serum then detected with fluorescent anti human globulin O 80 of patients have one or both autoAb s Q Addisons disease primary adrenal insufficiency O Autoantibody destroys the adrenal cortex type 2 hypersensitivity 0 Patients have insufficient cortisol production salt cravings and increased blood potassium O Diagnosed primarily by chemistry tests decreased cortisol response to ACTH adrenocorticotropin hormone injection 0 Type 1 diabetes pancreatic beta cell destruction 0 Type 1 diabetes mellitus old namelDDM results from autoimmune Tc destruction of pancreatic beta cells that normally secrete insulin type 4 hypersensitivity 0 Auto antibodies are detected before destruction of pancreas and disease symptoms hyperglycemia appear and appear to trigger disease initiation Q Ectopic lymphoid tissue can develop in pancreas 0 Diagnosis fasting hyperglycemia 0 Screening for predisease in research aimed at preventing disease onset includes combination testing for l AntiGAD glutamic acid decarboxylase antibody by EIA l AntilA2 transmembrane tyrosine phosphatase protein antibody by EIA l Insulin autoantibody lAAdone by immunofluorescence 3 Identify four autoimmune diseases that have a much higher incidence rate gt 5 times in women than in men 0 Hashimoto s thyroiditis G Graves disease 0 Systemic lupus erythematosis Q Addison s disease 4 Describe the general cause of Celiac Disease autoantibodies that can be used to diagnose it and why dermatitis herpetiformis is associated with this disease O The body produces antibodies which attack the intestinal tissue l lgA AntiGliadin l lgA antiendomysium attacks the smooth muscle of the intestine l lgA antiTTG Q AutoAntibodies used for diagnosis 0 lgA AntiTTG Tissue Transglutaminase O lgA AntiGliadin O lgA Antiendomysium Smooth muscle connective tissue 0 Dermatitis Herpetiformis Q If continue to eat gluten even though have gluten allergy you will get this Gluten sensitivity necessitating in a gluten free diet No wheat barley rye 0 Its associated with high Serum lgA AntiTTG Q Ulcerative colitis inflames the large intestine only and spares the anus O cANCA Antineutrophil cytoplasmic antibodies associated Q Crohn s disease can affect any part of the intestinal tract mouth to anus an is associated w skip lesions healtth and diseased tissue alternates Additional Information Antineutrophil antibodies ANCA are most commonly seen in ulcerative colitis patients who have vasculitis and another autoimmune vasculitis called Wegener s granulomatosis O Ulcerative colitis patients usually have a pANCA peripheral pattern 0 Wegener s granulomatosis patients usually have cANCA cytoplasmic pattern 6 Identify the autoantibodies associated with Pernicious Anemia and the vitamin deficiency they can cause 0 megaloblastic anemia due to cobalamin Vit B12 deficiency 0 antibodies interfere with uptake of cobalamin from intestine O antiintrinsic factor antibody is most commonly seen PA 0 antipariental cell antibody can also cause PA 7 Identify the autoantigen that is targeted in Myasthenia gravis the disease pathophysiology the immune mechanisms responsible for the disease and the laboratory test used for diagnosis Q Neuromuscular disorder with fatigue and skeletal muscle weakness 0 Due to Ab to acetylcholine receptor l Acetylcholine ACH is released from motor neuron ending and binds to ACHR on muscle fiber to initiate action potential I Ab blocks binding to ACHR and also increases receptor uptake by the muscle cell decreasing available receptor 0 Lab diagnosis by detection of antiACHR Ab by precipitation radioimmunoassay RIA 8 Identify the autoantigens that are targeted in Multiple sclerosis the disease pathophysiology and the immune mechanisms responsible for the disease 0 MS is an inflammatory autoimmune disease of the CNS 0 Autoantigens include I MBPmyelin basic protein l MOG myelin oligodendrocyte glycoprotein 0 These autoantigens are targeted by both Ab and T cells Th1 and Tc 0 Both To and antibody play a role in myelin destruction 0 Symptoms O Destruction of myelin sheath around axons causes visual problems weakness uncontrolled limb movement loss of balance and pinsneedles sensa ons 0 MS presents in two ways in affected patients I Chronic progressive disease I Acute relapsingremitting O No reliable lab test for diagnosis I MRI for lesions in brain or spinal cord used Topic 30 Autoimmunity Systemic Rheumatoogic Diseases Objectives 1 Identify the two most common rheumatologic diseases that are considered autoimmune diseases 0 Rheumatoid arthritis 0 Systemic lupus erythematosus SLE or Lupus 2 Explain why autoimmune rheumatologic diseases are difficult to diagnose Q Overlap of symptoms from one disease to another 0 Overlap of laboratory test results 0 No test is completely specific or sensitive for the disease ie not all patients have the expected autoantibodies 3 Define FANA ANA and ENA the staining patterns that may be observed in FANA and the two specific ENAs that are diagnostic for Systemic Lupus Erythematosus SLE orLupus 0 FANA Fluorescent antinuclear antibody usually SLE tests is performed by this 0 Patterns observed I Homogenouseven staining of entire nucleus most common I Speckled nucleus stains evenly except for centromeres I Nucleolar large discrete staining of nucleoli only I Peripheral nuclear rim staining outlines nucleus I Mixed a mix of above patterns 0 ANA Antinuclear antibodies classic screening test for SLE 0 ENA Extractable nuclear antigen 0 Anti dsDNA and antiSm are specific for SLE diagnosis but are NOT very sensitive I Anti dsDNA only positive in 5070 of SLE patients I AntiSmantibody to smith antigen only positive in 1530 of SLE patients I Anti histone ab is only ENA positive in drug inducedlupus 4 Categorize the autoimmune mechanism responsible for Lupus as one of the Type I IV hypersensitivity responses Type 3 hypersensitivity 5 Describe how titer is important in interpretation of FANA and what the maximum titer is that is still considered normal 0 Titer is important so that you don t get false positives Q lt 180 gt normal 0 gt 1160 gt positive 6 Identify other diseases that may show a low positive titer in an FANA test as well as other nondisease states 0 Positive FANA does NOT diagnose SLE O 5 of healthy ppl have low positive 0 Up to 70 of elderly have a low positive 0 Low positive titers in RA 50 o scleroderma 6090 Sjogren s syndrome 80 7 Identify the specific ENA that is associated with druginduced lupus 0 Antihistone antibody is only ENA in drug induced lupus though up to 70 of SLE patients have a low titer I Explain why lupus patients can have a false positive syphilis test Choose whether this false positive would be in either the nontreponemal or treponemal test for syphilis or both 0 False positive nontreponemal syphilis test 0 Indicator of anti cardiolipin antibody 0 Positive antidsDNA or antiSm Q SLE patients may develop antibodies to two phospholipids one of which can cause false positive syphilis test and is related with heart muscle inflammation anti cardiolipin The other is Lupus anticoagulant which binds to several clotting factors and phospholipid and falsely prolongs PT and APTT test but actually increases clotting in vrvo Q Anticardiolipin can cause false positive test results for syphilis This would be a nontreponemal test for syphilis 9 Define rheumatoid factor RF and identify what diseases it may be found in and for which disease it is considered most significant 0 Rheumatoid factor is lgM lgG or lgA antibody to the Fc portion of lgG antiimmunoglobulin antibodies 0 Most significant in RA 0 Also seen in syphilis lupus hepatitis TB Infectious mono and sjogrens syndrome 10 Describe how the screening test for lgM isotype RF differs from the specific tests for lgG and lgA isotypes of RF Which isare most specific for diagnosis of RA 0 Screening tests for RF are aggulination tests with either RBC or latex particles coated with human lgG 0 Only lgM RF will cause agglutination 0 NOT specific to RA 0 lgA and lgG tested by ElA or nephelometry are much more disease specific and present in nearly 100 of patients 11 Define the antigen used for antiCCP testing the disease this antibody is associated with and its usefulness in relation to RF testing 0 AntiCCP anticyclic citrullinated peptide is new screening test often paired with RF for new diagnosis of RA with 98 o specificity to RA 0 Citrulline is a non standard amino acid created by deamination of arginine by the enzyme peptidylarginine deaminase O This process is post translational modification of joint proteins fillagrin vimentin and fibrin makes them more durable I Self tolerance occurs in thymus or bone marrow where the post translational mod Are not made but if post trans Occurs in tissues then they are never gonna be exposed to antigen In patients with injury to joint and proteins are released to circulation and can be processed then you can get activation against that antigen 12 Identify the diagnostic criteria used for RA and whether it is necessary for the RF andor antiCCP tests to be positive or not 0 Criteria for RA diagnosis 6 or more points 0 Number of joints large and small involved up to 5 points 0 Seriology RF and AntiCCPACPA Up to 3 pts 0 Acute phase reactant testing Acute phase reactant testing up to 1 pt 0 Duration of symptoms up to 1 point 0 Of significance 0 RF and AntiCCP tests do not need to be positive to be diagnosed for RA 13 Recognize the laboratory tests that may be used to follow the course of SLE and RA and whether the results will be abnormally high or abnormally low in uncontrolled disease flares 0 SLE severity 0 Shows Increase in SLE l Rise of CReactive Protein Erythrocyte Sedimentation Rate AntidsDNA titer Urine Protein I Fall of CBC wbc rbc platelets Complement Serum albumin 0 RA progress 0 Increased severity of disease I Increased Erythrocyte Sedimentation Rate l Increased CReactive Protein l Decreased complement components TITER pg 35 Topic 31 Autoimmune Mechanisms amp Antibody Therapies Objectives At the completion of the modules and the assigned reading the student should be able to 1 Identify whether CD4 T cells CD8 T cells andor B cells are responsible for autoimmune disease a Both autoimmune antibodies B cells and cytotoxic T cells CD8 T cells have been found to be responsible for eliciting autoimmunity b All persistent autoimmunity must be due to breach in T helper cell tolerance c HENCEthe link of autoimmunity to HLA genes especially MHC class II but also MHC class I d So all of them are responsible 2 Identify two proteins involved in self tolerance that when defective in humans lead to generalized autoimmune disease a Homozygous deletion mutations of i AIRE ii FoxP3 b Or lL1O and TGFB functional cytokines of Treg 3 Explain how an allele of CTLA4 normally found in the human population can lead to increased susceptibility to autoimmune disease a One allele of CTLA4 limits production of soluble CTLA4 which is apparently important in optimal Treg function i WEAK risk factor ii Allele is present in 50 o of population increases to 60 in patients with autoimmune disease iii NOT associated with increased risk for a particular autoimmune disease but instead autoimmunity in general iv Suppression of autoreactive T cells by Treg cells requires them to interact with the same APC 4 Define relative risk and the ultimate effect that it has on an individual s likelihood to develop any given autoimmune disease Be able to use disease incidence and relative risk to calculate actual risk for an individual who may have an at risk allele a Relative risk RR if you have the associated HLA type RR is the increased risk you have over the general population to get the disease most are MHC class 2 b If you have the HLA susceptibility it does NOT mean you will get the disease i If total risk population is 1 in 100000 ii Your relative risk is 35 due to HLA association iii Your actual risk is 35 in 100000 or 1 in 28600 10000035 iv 128600003 o chance 1 RR1 no association 2 RRgt1 positive association 3 RRlt1 is negative protective association 5 Explain the role of HLA alleles in development of autoimmunity ie what are the HLA molecules doing exactly to trigger autoimmunity a Human HLA MHC class 1 and 2 are linked to increased or decreased susceptibility to certain diseases b Fails to provide specific protection c Presenting self antigen d People with certain HLA Ag are more likely to develop certain autoimmune diseases 6 List and explain the mechanisms of at least six 6 factors that have been proposed to be risk factors for autoimmune disease development besides gender and HLA alleles In doing so be sure to define the term hapten a Release ef sequestered antigens tissu a damage Narmally intracellular preteins and cleic aCidiS not in uid certain are red in certain mquot Traumatic mayquot release these molecules and at the same time elicit an inflammatory response 10 Inflammation causing ectoie expression of on cells that weuld normally not express it IFN gamma can cause several cell types to express class ii cells a protein not in the thymus an hence no T cell tolerance exists to the prete in iii However the cells still do not usually have eo stimulatory molecules 0 Molecular mimicry in an une response to a pathogen trigge a R that crass reacts self Ag i Rheumatic fever Ab to group strep bi ndrs t heat protest cernlement activation and acute inflamm atory reapense against the heart uscle usu ally Itf limit e ii Coxsaekie virus proteinsAb made in response to cross reaet with pancreatic eta cells and may increase risk or tye 391 diabetes at Polyclonal b cell activation can trigger Ab to Ag i negative bacteria and EBV all polyclenal cell ii Antibedies produced are lg M no T cell help 6 Plant mitogens cause paol yC39lonal lymhocyte activation r Miner defects the itself Inhibitory F cyiR polymorphis lead to inability to inhibit B cell even antibedy adequate ii Genetic mutations in Ct C4 and mplement receptors that bind 1 1q predispose to SLE 1 Hlyip ethzeticaxl explanation in effective clearance of immune lexes eventually activates complement where the Ab deposits in tissues cells instead clean and neat apoptosis eut cell proteins which are processed and presented in MHC class 2 g Chemical Environmental i Chemicals pollutants smoking tobacco food toxins 1 Can act as a haptensomething that by itself cannot act as an antigen in the immune system but when it is chemically covalently linked to our own protein it now produces a new novel antigen that you can respond to ex Poisen ivy to change self proteins into novel Ag 2 Can cause chronic damage that releases hidden Ag ii Once the immune response is triggered the BCRTCR then recognize the normal self protein as foreign as well 7 Define epitope spreading in the context of autoimmune disease and how linked recognition is responsible for epitope spreading Take what you want an antibody response to and inject it into a large animal many times over an extended time period to create a response and then take the serum out a When a patient is first diagnosed auto Ab are limited b As the autoimmune disease progresses the patient develops autoAb against more and more autoAg c This is thought to be due to the sloppiness of T cell help to B cells d CD4 t cells specific for one epitope of a macromolecular complex can provide help to B cells specific for other accessible epitopes of the complex e A B cell that internalizes a macromolecular complex can present Ag to T cells specific for any of the proteins in the complex To get t cell help and present Ag f Bcr and tcr have linked recognition and it can recognize them 8 Discuss the general usefulness of Mg RhoGAM and other antibody preparations used in the therapeutic treatment of humans 0 Used in human medicine 0 Mg concentrated human IgG pooled from multiple normal donors 0 RhoGAM used in prevention of hemolytic disease of the newborn poly clonal antibidy purified from pooled Rhnegative Mom s who have anti Rh antibody 0 monoclonal antibody or recombinant protein therapies used to target and kill tumors or as immunosuppressants 9 Describe the general process of hybridoma creation for the production of monoclonal antibodies 0 Monoclonal antibodies are antibody preparations in which the antibody is all identical produced by a cell clone called hybridoma grown in vitro 0 Production 0 Inject immunize animal mouse with an antigen so the animal will make antibody to the antigen 0 Isolate B cells plasma cells from the spleen of the animal and fuse the B cells to an immortalized myeloma o Grow fused cells in chemical selection media so that ONLY cells that have fused B cell and myeloma can survive unfused spleen cells and myeloma cells will die 0 Singlecell clone the surviving hybridoma cells and select for a clone that produces high affinity antibody against the antigen you used to immunize the animal your target antigen 0 Isolate the pure monoclonal antibody from this hybridoma cell line 10 Describe how monoclonal antibody preparations differ from polyclonal antibody preparations aka antiserum 0 Reagents for test kits clinical laboratory testing and research immunohistochemistry clinical chemistry nephelometry testing EIA flow cytometry virtually any test that relies on antigen identification using an an body Q Treatments for infusion into humans exploding market 0 Poly Clonal l Mixture of different isotypes and different affinities l Good for treatment 0 Monoclonal l One single isotype with one single affinity l Good for diagnosis 11 Identify specific monoclonal antibody therapies used to treat Rheumatoid Arthritis and other destructive autoimmune disorders 0 Treatment of specific cancers 0 lmmunosuppressive uses of mAb O AntiTNFalpha blocking mAb used to treat rheumatoid arthritis blocks joint destruction 0 Antialpha4 integrinblocks homing of inflammatory cells preventing autoimmune tissue destruction MS IBD psoriasis O AntilL1 or antilL6 antibodies to interfere with macrophage inflammatory cytokines when antiTNFalpha does not work 12 What are HAMA Would they cause false positive of false negative assay results in clinical laboratory EIA testing 0 HAMA Human antimouse antibodies 0 Generally cause a false positive result that does not correlate with patient history 0 Problems with monoclonal antibody therapies 0 Any laboratory assay that uses enzyme immunoassays for detection and quantification of an analyte can give false positives if antibodies against another species are present Commonly called Heterophile antibodies pg 36 Topic 32 Vaccines Topic 32 Vaccines Objectives At the completion of the modules and the assigned reading the student should be able to 1 Define the following terms antigen immunogen hapten adjuvant vaccination herd immunity 0 Antigen a substance that reacts with Ab BCR or T lymphocytes TCR 0 lmmunogen implies that the Ag evokes an immune response by causing inflammation 0 Hapten small molecules even chemicals that become immunogenic when complexed to larger molecules like proteins even though by themselves they are not immunogenic 0 latex allergies drug allergies penicillin poison ivy are all examples of this 0 Adjuvant a substance administered with mixed with an antigen to enhance the immune response to the antigen 0 vaccination any deliberate immunization intended to induce protective immunity against disease 0 herd immunity the ability to prevent epidemics of dangerous infections when the vast majority of individuals are immune by vaccination 0 used this to wipe out smallpox 2 Identify the 4 general requirements necessary for a Good antigen an antigen that will be immunogenic 1 Must have a high molecular weight at least 100000 2 Should have high molecular complexity like proteins made of 20 different amino acids 0 Carbs and lipids gain molecular complexity and become more immunogenic as glycoproteins or lipoproteins 3 Foreignness taxonomically distant from the host 4 lnflammatory usually by possessing TLR ligand or inducing cell stress 3 Identify 4 mechanisms that may be responsible for the ability of adjuvants to enhance the immune response to vaccination 1 Complex with the antigen to increase its size 2 Trap the antigen in material that is relatively indigestible so anitgen slowly leaks out over time lipid emulsion 3 Hyperactivate the innate immune cells like macrophages most likely via TLR s 4 Modulate the immune response by favoring Th1 vs Th2 or vice versa 4 Identify the types of vaccines that are currently in use to immunize individuals against viral infections and bacterial infections including any limitations of the individual types that we may have discussed in class Viral vaccines are one of 3 tvnes Killed or inactivates vaccines O Treated with formalin heat or irradiation so that they are no longer able to replicate 0 Used for salk polio vaccine influenza and rabies Live attenuated viral vaccines 0 More potent than killed vaccines because virus replicates and mimics real diseass 0 Virus is grown in culture on nonhuman cell line and it mutates to grow better on these cells but worse in human cells so that it helps you not infects you cant grow well in humans 0 Used for sabin polio vaccine measles mumps 0 Subunit vaccines 0 Use purified or recombinant surface components or viral capsidenvelope to induce neutralizing antibody never use alive attenuated vaccine for HBV 0 Currently used to hepatitis B virus vaccine with alum to induce protective antiHBs Ab 0 Only effective in 85 of population those who do not respond likely lack a MHC class 2 peptide epitope to present to T cells 0 Also used for hepatitis A vaccine Bacterial vaccine strategies 0 Live attenuated bacterial vaccines 0 Previously created by selection of lesspathogenic strain of bacterium BCG is attenuated mycobacterium bovis used to vaccinate humans for TB 0 Currently created by molecular biology in which a protein necessary for pathogenesis is intentionally mutated by molecular biology 0 Used for salmonella typhi typhoid fever vaccine 0 Bacterial subunit vaccines can be one of two types 0 inactivated toxin vaccines toxoids l toxin is purified treated with formalin to destroy toxicity but retain antigenicity I used for diphtheria and tetanus I must be combined with another vaccine that is more inflammatorywhole killed bacteria to be effective DTP has whole pertussis bacteria newer DTaP has acellular pertussis antigen mix 0 purified capsular polysaccharide vaccines l polysaccharide normally protects bacteria by inhibiting C binding but antibody to polysaccharide can induce 0 binding and lysis I Durified polysaccharide only induces protective Tindependent antibody in healthy adult I coniuoate vaccines couple polysaccharide to a Tdependent protein anitgen to stimulate CD4 helper t cell response and protection in young children and elderly iClicker Questions Lecture 26 1 Which stage of syphilis can present with a painless ulcerated lesion with red raised borders usually on the genitals a Primary Syphilis 2 ln nontreponemal tests for syphilis the reagin antibody in the patient serum if present is binding to a Cardiolipin 3 Charcoal particles are a component of the test reagent for which test for syphilis a RPR 4 The causative agent of lyme disease is a Borrelia burgdorferi 5 The confirmatory test for Lyme Disease is a idk western blot pretty sure Lecture 27 1 Group A streptococcus is a a Gram Positive Coccus Organism b Grows in chains 2 A super antigen causes pathology in the body by a Activating T lymphocytes with common VB gene segments by holding them to mhc 3 Acute proliferative glomerulonephritis can follow which primary infections with group a Strep a Group A strep Pharyngitis or lmpetigo 4 Which test is known to be negative following true cases of Streptococcal Pyoderma a ASO Test 5 Which secondary Group A strep Sequelae results from cross reactive antibodies binding to muscle components especially heart muscle a RHumatic Fever Lecture 28 1 Type IV hypersensitivities are caused by what immune mechanisms a CD4CD8 T lymphocytes 2 An immediate hyperssensitivity is the same as a a Type I Hypersensitivity 3 Histamine is responsible for many of the symptoms in a Type I hypersensitivity 4 What is a disadvantage of skin prick testing for allergies a Testing is traumatic to patient because each allergen tested uses a needle prick b Patient must discontinue use of antihistimine for 48 hours before hand c Potential of producing anaphlaxic shock 5 Serum sickness can be a symptom of a severe form of which hypersensitivity a Type III Lecture 29 1 Most autoimmune diseases are associated with what type of hypersensitivity to self antigen a Type II 2 Hasimoto s disease chronic Autoimmune Thyroiditis is due to what type of hypersensitivity a Type IV 3 Which of the following diseases results from autoimmune destruction of the pancreatic B cells by To a Type 1 Diabetes 4 Which of the following is an autoimmune diseases that results in cobalamin deficiency a Perinocious Anemia 5 Myelin Basic Protein is one of the two most common aautoantigen targed in which autoimmune disease a Multiple Sclerosis Lecture 30 1 In acute disease flares of systemic lupus which of the following assays will have a higher test result in patient a CReactive Protein Complement C3 would be lower 2 Which of the following antidsDNA titers would correlate with more severe disease in a patient previously diagnosed with lupas a 1160 or 1 2560 i 12560 3 AntiCCP autoantibody is associated with which autoimmune diseases a Rheumatoid Arthritis 4 The agglutination test for Rheumatoid factor RF is specific for rheumatoid Arthritis diagnosis a False 5 The RF Test must be positive in order for a person with appropriate symptoms to be diagnoised with rheumatoid arthritis a False Lecture 31 1 WHich of the following immune cell hashave been shown to cause autoimmune disease a CD4 and CD8 and B cells 2 Which of the following proteins is associated with autoimmunity when it is mutated absent or somehow reduced in function a FoxP3 3 HLA allel has a calculated risk of 2 for grave s disease That means you have the allele you have a risk of developing the disease compared to general public a Lower 4 Epitope spreading in autoimmune disease is a Can be attributed to sloppy linked recognition between b and t cells b Is responsible for chronic worsening of autoimmune disease the longer someone has it 5 Monoclonal antibodies paeparations are mixtures of antibodies and affinities a false Lecture 32 1 What percent of individuals in a population need to be protected by vaccination in order for herd immunity to exist for that pathogen a 95 2 A small reactive chemical that can covalently attach to proteins and thereby become immunogenic would properly be called a A hapten 3 What strategy can be employed with adjuvants to make an antigen more immunogenic a Adding a TLR Ligand 4 Viral subunit vaccines are generally used when a A virus is too dangerous to risk to use of liveattenuated or chemically inactivated virus 5 What is a disadvantage of a live attenuated viral vaccine a It can be dangerous for someone who is immunosurpressed
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