Week 2 notes- Immunology
Week 2 notes- Immunology Bio 5500/5600
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This 14 page Class Notes was uploaded by Mary on Sunday August 30, 2015. The Class Notes belongs to Bio 5500/5600 at Auburn University taught by Dr. Schwartz in Fall 2015. Since its upload, it has received 58 views. For similar materials see Immunology in Biology at Auburn University.
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Date Created: 08/30/15
Class BIO 55006500 Pattern Recognition Receptors Recognition of microbes and damaged self by the innate immune system Pathogenassociated molecular patterns PAMPs and Damageassociated molecular patterns DAMPs PAMPs recognize molecular structures produced by microbial pathogens 0 Different type of microbes will express different PAMPs DAMPs are endogenous molecules that are associated with damage that results from an injury that should be recognized by the immune system 0 DAMPs can also indicate a sterile injuries acquired through burns and chemical toxins 0 Cell recognition molecules of the innate immune system are expressed by phagocytes primarily macrophages and neutrophils dendritic cells and epithelial cells that form barrier from the body to the environment 0 Patter recognition receptors PRR are receptors for pathogens and damageassociated molecules 39 PRRs recognize patterns 0 PRR bind to PAMPs and DAMPs activating signal transduction pathways which in turn promotes antimicrobial and pro in ammatory functions of the cell in which they are expressed O PRR is the larger term used for tolllike receptors Cellassociated pattern recognition receptors PRR and sensors of innate immunity Tolllike receptors recognize products of a wide variety of microbes and molecules expressed by stressed or dying cells Tolllike receptors were first identified and named after the Drosophilia fruit y receptor Toll discovered by Jules Hoffman and colleagues in 1996 o It was found that the toll protein also mediated antimicrobial responses in fruit y There are 9 different functional TLRs in humans o TLRs 19 Endosomal TLRs Ligand Microorganism recognized TLR3 dsRNA Viruses west niles TLR7 ssRNA Viruses HIV TLR8 ssRNA Viruses in uenza TLR9 Unmethylated Cpg DNA Bacteria viruses herpes TLRs 378 and 9 are mainly expressed inside cells on the endoplasmic reticulum and endosomal membranes Plasma membrane TLRs Microorganism L1gand recogmzed TLR1TLR2 heterodimer Bacterial lipopeptides Bacteriaparasites TLRZ Bacterial peptidoglycan Bacteria TLR4TLR4 homodimer Lipopolysaccharide LPS Gramnegative bacteria TLRS Bacterial agellin Mot11e bacter1a havmg a agellum TLR2TLR6 heterodimer Bacterial 1i 0 e tides Gram positive p p p bacteriayeast fungi TLRs 1245 and 6 are expressed on the plasma membrane where they recognize different PAMPs in the extracellular environment 0 Tolllike receptors pair as they bind ligands and signal 0 TLRZ binds to either TLR 1 or 6 39 With each one it changes the specificity of binding 0 TLR 4 needs help with binding to its ligand lipopolysaccharide 39 It has to be extracted from bacterial membranes or liposomes and it has several molecules that work with it to make LPS available for recognition 0 Myeloid differentiation protein MDZ CD14 and lipopolysaccharide binding protein TLRs can form homodimers or heterodimers o TLR 4 and 2 form homodimers The specificity of the TLR system is extended by the ability of TLRs to heterodimerize with each other 0 ie Dimers of TLRZand TLR6 are required for response to peptidoglycan o Ligand binding to the leucinerich domains cause physical interactions between TLR molecules and the formation of TLR dimers Pattern recognition Location Specific Example PAMP DAMP Ligands receptors Cellassociated Plasma membrane and Varlous m1crob1al molecules TollLike endosomal membranes of including bacterial LPS and receptors dendritic ells phagocytes B TLRs 19 td 1 cans iral n Cleic TLRs cells endothelial cells and pep 1 0g y Civ u others an S Bacterial cell wall peptidoglycans 1123113615 Cytosol of phagocytes NLlliglgal ily Intracellular crystals urate NLRS epithelial C8115 and Others in ammasomes silica changes in cytosolic ATP and ion concentrations lysosomal damage Table 1 Patter recognition molecules in the innate immune system These molecules are responsible for facilitating the clearance of microbes from blood and extracellular uids by enhancing the uptake in phagocytes or by activating extracellular killing mechanisms Innate immune receptors are only able to tell damaged cells from healthy cells 0 They are not able to tell particular species of microbes or cell types Tolllike receptors have ligand binding domains and transmebrane regions NODlike receptors since they are cytosolic they do not have transmembrane regions because they exist in they cytosol outside a membrane bound compartment Class BIO 55006500 TLR recognition of microbial ligands results in the activation of several signaling pathways and ultimately transcription factors that induce the expression of genes that are important for in ammatory and antiviral responses Initiated by hgand binding to Expression of Acute TLR leading NF KB is in ammatory in ammation to Egg activated by genes amp stimualtion dimerization TLR signaling cytokines of adaptive of TLR chemokines immunity roteins TLR12456 Q g Q g a a a a a Initiated by ligand binding to Expression IRFs are lealfnl gm e355 Egt agtivgglgd i ietryf ioln 555 Egt aggigggal dimerizatio with TRIF 3 ln IFN aB INFs n of TLR 51g a 1 g genes proteins TLR34 K K K K g Figure 1 Signaling pathways and functions of TLRS LPS activates TLR4 Peptidoglycan activates TLR1 256 In general MYD88 dependent signaling is the major pathway through which you get acute in ammation and the production of proin ammatory cytokines NFKB Transcription factor is important because its at the center of almost every immune response 0 It is the transcription factor that regulates most genes in in ammatory responses 0 A very critical molecule Type1 interferons good at fighting viral infections Think TRIF and interferon O Trif you get antiviral response 0 Transcription factors interferon regulatory factor that regulates the production of type 1 interferon TLR 124567and 9 signal through MYD88 0 When you think of MYD88 think of Pro in ammatory response Know link between toll like receptors MYD88 NFKB and pro in ammatory cytokines Mouse Ips gene Mutaion maps to TLR4 Beutler and colleagues in 1998 reported in Science an actual function for TLR4 and its role in the LPS response using the endotoxinresistant C3H HE mice These mice were found to have a point mutation in the TLR4 gene that was attributed to a loss response to LPS Endotoxintolerant mice have mutations in Tolllike receptor 4 TLR4 Qureshi ST Lariviere L Leveque G Clermont S Moore K Gros P Malo D I EXP Med Feb 1999 18961525 0 Give a normal mouse a bolus of LPS they develop septic shock and it is fatal 39 Tolllike receptor 4 was the deficiency in the mice 0 C3H He strain had very little detectable response to the injection 39 Identified TLR 4 was the deficiency attributed to the lost of response to LPS Cytosolic Receptors for PAMPs and DAMPs Three major classes of these cytosolic receptors 1 NODlike receptors nucleotide oligomerization domaincontaining protein 0 Generally recognize bacterial cell wall components 0 The NLRP subfamily of NODlike receptors respond to cytosolic PAMPS and DAMPs by forming signaling complexes called in ammasomes O In ammasomes hook u with machinery know as caspase that are essential for generating biologically active IL 1 and IL 18 39 Initially expressed as pro IL1 or pro IL 18 0 Promote in ammation 2 RIGIlike receptors and MDAS o Recognize viral nucleic acids 0 Produce type 1 interferon when RLRs bind viral RNA leading to activation of IRF3 and IRF7 as well as NFKB which induces the production of type 1 interferon 3 Cytosolic DNA sensors 0 Promote in ammation or type 1 interferon production Downstream effects of signaling Downstream effects fever swelling IL 6 production and acute phase response as well as dendritic cell maturation NFKB acitvates gene transcritpion These genes encode o proin ammatory cytokines IL6 IL1 TNF IL 12 and IFNy o chemokines such as IL 8 0 Lipid mediators such as prostaglandins leukotrienes and platelet activating factor PAF o Anitin ammatory cytokine such as IL 10 and TGFB Activation of IRFs such as IRF3 and or IRF7 leads to production and secretion of type 1 interferons Class BIO 55006500 These proin ammatory cytokines chemokines and other mediators orchestrate the in ux and control of neutrophils macrophages dendritic cells and other leukocytes such as T cells and B cells that enter the tissue site of infection Localized vs Systemic infections 0 A little in ammation is a good thing 0 At the heart of septic response is vascular permeability 0 When you have a lot of vascular permeability you cant have decent blood pressure you cant properly oxygenate your organs and you have organ failure Class BIO 55006500 The Complement System The complement system consists of several plasma proteins that work together to opsonize microbes to promote the recruitment of phagocytes to the site of infection and in some cases to directly kill microbes Activation involves protelytic cascades in which an inactive precursor enzyme zymogen is alters to become an active protease that cleaves inducing the activity of the next complement protein in the cascade Soluble recognition receptors and effector molecules of innate immunity There are several molecules that recognize microbes and promote a innate response that exist in a soluble form in blood and extracellular uids 1 Control in ammation recruitment of phagocytes Z Enhance pathogen uptake and clearance Opsonization o Pathogens become decorated with complement proteins and bind with specific factors Opsonization 3 Lytic attack of cell membranes Killing bacteria 0 Pore formation on bacterial membranes The Complement system Pathways Trigger Downstream effect Deficiencies Classical Pathway Antigen antibody Clr and C15 are formed Deficiencies in MAC complexes and initiate proteolytic causes increased Clq and pentraxins can activate cascade Perforation of pathogen ell membranes susceptibility to a limited of microbes ie Neisseria bacteria Deficiency in C1 will cause deficiency in classical pathway Alternative Pathway Pathogen surfaces C3 gets broken up into Deficiencies in C3 First to act C3a and C3b cause for increased C3 complement susceptibility to protein Recruitment of reoccurring and often in ammatory cells lethal bacterial No initiation infections molecule MBLectin Pathway Lectin binding to Two zymogens formed Deficiency in MASP 1 pathogen surfaces Mannosebinding lectin MBL Highly specific for pattern of molecules MASP 1 and MASP 2 associate with MBL to initiate proteolytic steps identical to classical pathway Opsonization of pathogen and MASP 2 will cause a deficiency in MB Lectin pathway All three pathways result in recruitment and assembly of different complement proteins into protease complexes One of these complexes is C3 convertase that cleaves C3 which then produces C3a and C3b 0 C3 convertase in the classical pathway is made up of C2a and C4b in the alternative pathway it is made up of C3b and Bb o C3a leads to recruitment 0 C3 convertase is at the heart of all three pathways which eventually leads to increased permeability migration of monocytes and neutrophils into the tissue 0 C3b serves as an opsonin to promote phagocytosis of the microbes I C3b can act as receptor and bind to bacteria leading to opsonization o The key feature in optimization is the zippering of ligands and pulling into the cell 0 C3b also binds to other complement proteins forming a protease called C5 convertase I C5 convertase cleaves C5 and forms C5a and C5b C5 is a very active anaphylatoxin very good at recruiting cells I C5a causes in ammation I C5b initiates the formation off C6 C7 C8 and C9 which are assembled into a membrane pore called the membrane attack complex MAC C3a is released and stimulates in ammation by acting as a chemoattractant for neutrophils Regulatory proteins of the classical and alternative pathways C1 inhibitor CIINH binds to activated C1r Cls removing Clq and to activate MASPZ removing it from MBL Factor H H Binds C3b displacing Bb cofactor for I Factor I I serine protease that cleaves C3b and C4b aided by H MCP C4BP or CR1 CD59 protectin prevents formation of membrane attack complex on autologous or allogeneic cells Widely expressed on membranes 0 Factor H and I are intermediate stages 0 CD59 binds to C9 and prevents formation of the pore How does the alternative pathway differ from the other two pathways Lacks initiation molecule 0 Comes from activation of complement directly on bacterial surface 0 Happens spontaneously at all times constantly being hydrolyzed so C3a and C3b are being produced 0 C3 is spontaneously hydrolyzed in serum and the end result is C3b which can attach to the bacterial membrane and liberation of C3a which acts as a chemoattractant anaphylatoxin to phagocytes 0 Self perpetuating Class BIO 55006500 The Terminal complement components that form the membrane attack complex Protein Concentration Function in serum Hgm1 C5 85 On activation the soluble C4b fragment initiates assembly of the membraneattack complex C6 60 Binds to and stabilizes C5b Forms a binding cute for C7 C7 55 Binds to C5b6 and exposes a hydrophobic region that permits attachment to the cell membrane C8 55 Binds to C5b67 and exposes a hydrophobic region that inserts into cell membrane C9 60 Polymerization on the C5b678 complex to form a membranespanning channel that disrupts the cells integrity and can result in cell death Everything from C5b is specific to the membrane attack complex Membrane lytic component part does opsinization Pentraxins Family of penatmeric proteins in plasma that recognize microbial structures Prominent membranes of the penatmeric family 0 Creactive protein CRP I Bind to several species of bacteria and fungi I Bind phosphorylcholine an acute ohase protein indicator of infection and in ammation I Recognizes the molecular ligand phosphatidylethanolamine o Sserum amyloid P SAP I Bind to several species of bacteria and fungi I Binds phosphatidyl ethanolamine I Recognizes the molecular ligand phosphatidylethanolamine 0 Long pentraxin PTX3 CRP SAP and PTX3 bind Clq and initiate the classical pathway CRP plasma concentration are normally very low in healthy adults however increase up to 1000 fold during infection known as acute phase reactants since they are high in the blood during in ammatory reactions 0 This increase is a result of increased synthesis by the liver induced by cytokines IL 6 and IL 1 Collectins and Ficolins Collectins a family of trimeric or hexameric proteins Contains a collagenlike tail with Ctype lectin head Collectin soluble effector molecules of the innate system 0 Mannosebnding lectin MBL I Binds mannose bacteria and initiates MLB pathway I Also works as an opsonin by binding to and enhancing phagocytosis 0 Pulmonary surfactant SPA and SPD I Main purpose is to maintain the abilities of the lungs to expand and as a mediator of the innate immune responses in the lung I Reside in the alveoli I Act as opsonins with some direct killing activity I Defect in surfactant will lead to increased bacterial diseases Ficolins plasma proteins structurally similar to collectins Conatin collagenlike domain but have a fibrinogen domain instead of a C type lectin domain 0 Binds opsonizes and activates complement in a manner similar to MBL The In ammatory response Three major ways in which the innate immune system protects against infection 1 Inducing in ammation 2 Inducing antiviral defense 3 Stimulating the adaptive immune system One of the earliest responses to the innate immune system to infection and tissue damage is the secretion of cytokines by tissue cells 0 The major proin ammatory cytokines are IL 1 and IL 6 0 Cytokines of the innate immunity are produced mainly by macrophages and dendritic cells 0 Act on cells close to their cell of orision 0 Can have similar and overlapping actions or are functionally unique 0 One cytokine can stimulate the development of another amplifying the reactions 0 Induce in ammation inhibit viral replication promoting T cell responses and limiting innate immune response Three of the most important cytokines of the innate immunity are 1 TNF mediator of the acute in ammatory response to bacteria and their infectious microbes O Produced by macrophages dendritic cells and other cell types 0 Causes necrosis of tumors as a result of in ammation and thrombosis 0 Causes septic shock 2 IL1mediator of the acute in ammatory response with similar actions as TNF 3 IL6 mediator of the acute in ammatory response having both local and systemic effects 0 Induces the synthesis of other in ammatory mediators in the liver stimulates neutrophil production in the bone marrow and promotes the differentiation of IL 7 producing helper T cells 0 TNF and IL 1 and IL 6 are responsible for many of the clinical signs of infections and in ammatory disease Class BIO 55006500 0 Act on hypothalamus to induce fever and increase body temp endogenous pyrogens IL12 secreted by dendritic cells and macrophages and stimulates the production of IFNA production by NK cells and T cells 0 IL 12 is a heterodimer made up of 31 and 32 subunits Ingestion and killing of Microbes by Activated Phagocytes Neutrophils and macrophages are recruited into site of infection Ingestion and killing of Microbes by Activated Phagocytes covered in depth on 82 1 notes I Phagocytosis active energy dependent process of engulfment of large particles Microbe binds to phagocytic receptors 9 phagocyte membrane zips up around molecule microbe ingested in phagosome fusion of phagosome with lysosome 9 activation of phagosome killing of microbes by ROS N0 and lysosomal enzymes in phagolysosomes Phagosome contains the ingested foreign particle Reactive oxygen species ROS activated macrophages and neutrophils convert molecular oxygen onto reactive oxygen species ROS 0 Highly reactive oxidizing agents that destroy microbes Nitric oxide macrophages produce reactive nitrogen species mainly nitric oxide NO by the action of enzyme called uinducible nitric oxide synthesis iNOS Proteolytic enzymes produced by activated neutrophils and macrophages Produced in the phagolysosomes that function to destroy microbes Neutrophil extracellular traps NETS neutrophils kill microbes by extruding their DNA and granule contents forming extracellular threads net on which bacteria and fungis are trapped and killed Symptoms and Pathologic Consequences of In ammation TNF and IL 1 act on the hypothalamus to induce fever 0 These cytokines are known as endogenous pyrogens Fever is induced by increasing the production of prostaglandins in hypothalamic cells IL 1 and IL 6 induce hepatocytes to produce acute phase reactants CRP SAP and fibrinogen which are secreted into the blood 0 CRP and SAP play protective roles in infection 0 Fibrinogen contributes to hemostasis and tissue repair TNF has many roles in severe infections 0 Plays 0 Inhibition of myocardial contraction and vascular smooth muscle tone resulting in marked decrease in blood pressure or shock Intravascular thrombosis Stimulates endothelial cell expression factor activates coagulation Inhibits expression of thrombomodulin which inhibits coagulation 0 Causes necrosis of tumors Antiviral response Type 1 interferons a large family of structurally related cytokines that mediate early innate immune response to viral infections Class BIO 55006500 Innate Immunity IV Antiviral Response Acts in both autocrine and paracrine manner to protect from further viral infection 0 Autocrine supporting of self 0 Paracrine helping your neighbors Induction of type 1 Interferons in response to viral infections Produces Interferon B and several interferon a s o IFN 3 is produced by many cell types 0 Major source of IFN a is plasmacytoid dendritic cells but can also be produced by mononuclear phagocytes I Both IFN 3 and IFN a bind to the IFN a receptor 0 Type 1 Interferon a large family of structurally related cytokines that mediate the early innate immune response to viral infections 0 Primarily a paracrine action in that a virally infected cell secretes interferon to act on neighboring cells that are not yet affected 0 Increase expression of ligands for receptors on NK cells 0 Increases the toxicity of NK cells and promotes the differentiation of naive T cells to the TH1 subset of helper T cells 0 Induces resistance to viral replication in all cells 0 Upregualte the expression of class 1 MHC molecules increasing the probability that virus infected cells will be recognized and killed Biological actions of type 1 interferons 0 Production of type 1 interferon by virally infected cells induces an expression of enzymes that block viral replication antiviral state and lead to 0 Inhibition of viral protein synthesis 0 Degradation of viral RNA 0 Inhibition of viral gene expression and virion assembly shuts down the virus expression 0 Downstream effect review 0 Viral recognition 0 Production of interferon B and several interferon a s 0 Induction of resistance to replication antiviral response 0 Increase the expression of ligands fro receptors on NK cells 39 Can upregualte MHC making it a better target for a T cell 0 Induced heightened expression of things that activate the NK cells Tolllike receptor 0 TLR 78 and 3 have special specificity for certain viral DNA RIGlike receptors cytosolic sensors of viral RNA that respond to viral nucleic acid by inducing the production of antiviral type1 interferon 0 Two RIGI receptors RIGI and MDA5 recognize different sets of viral RNAs that are characteristic of distinct viruses Interferon regulated factor RIF transcription factor that regulates interferon responses 0 Involved in type 1 interferon synthesis and its regulating genes Stimulation of Adaptive Immunity The innate immunity response provides signal that function in concerti with antigens to stimulate the proliferation of antigenspecific T and Blymphocytes The innate immune response provides the initial defense against microbes and is the precursor to the adaptive immune response Natural Killer Cells and Other Innate Lymphoid Cells Natural killer cells the best described innate lymphoid cell Lymphocytes if the innate immune system that play an important role in innate immune response mainly against intracellular viruses and bacteria Express receptors that recognize altered or normal ligands on target cells Major function killing infected or quotaberrantquot injured cells and production of IFNy o IFNy activates macrophages to destroy phagocytosed microbes 39 Type 1 interferon alpha and beta 11 alphas interferons and 1 beta interferon 0 Viral clearance 39 Type 2 interferon gamma 1 0 Product of NK cells and T cells 0 Activates macrophagesmakes macrophages a better killer of the pathogens it has already taken up Killing activity is balanced by activating and inhibitory receptors 0 Forms a quotsynapsequot between immune cells and delivers toxic molecules to site of interaction 0 Ultimately result in DNA cleavage nuclear fragmentation membrane blebbing Use germline DNAencoded receptors to distinguish pathogen infected cells from healthy cells 0 Healthy cells express ligand MCH class 1 0 When activating receptor is triggered a signal is transferred to NK cell 0 Depends whether it is positive or negative signal whehter the Nk cells kills it 39 If there is an aberrant protein giving a very strong positive signal it will result in killing of cell of the cell 0 If you have both inhibitory and activating ligand being expressed from a cell if the signal from the inhibitory receptor is stronger the NK cell will not kill it It will be left alone 0 If a cell is expressing aberrant proteins and has down regulated MHC class 1 it is a clear situation on which NK cell would kill it 0 If there are no aberrant proteins expressed and MHC is at normal levels the cell will not be killed straight forward scenario 0 Perforin a NK cell granule facilitates the entry of other granules granzymes into the cytosol of target cell A poreforming molecule 0 Granzymes initiates signaling that leads to the death of the target cell through apoptosis act through capases 39 They are delivered through the pore Class BIO 55006500 Importance of NK cells 0 They act in the window prior to the adaptive response kicking in O Regulate viral replication during this time 0 When cells are infected by viruses they often downregulate molecules such as MHC which are recognized by T cells 0 When cells are infected especially by viruses they often express stress proteins so NK cells fill the gap and respond to the cells that express stress proteins and have lost to expression of normal molecules Virally infected cells Can cause their own proteins to be expressed which would be recognized as different 0 Cause normal host proteins to be aberrantly expressed misfolded 0 Can down regulate MHC class 1 0 It makes the cell invisible to cytolytic T cells which need MHC call 1 to recognize and kill their targets Activating and Inhibitory Responses of NK Cells Killer cell immunoglobulin Ig like receptors KIR s contain a signal domain Ig fold KIR s can be activating or inhibitory depending on their cytoplasmic tail 0 Inhibitory receptors have long cytoplasmic tails 39 Stimulates phosphatases that counteract kinase 39 Contains inhibitory motifs inside cytoplasmic tails that bind to phosphatase and truncate a signaling cascade 39 Binds to normal class 1 MCHlike proteins 0 Activating receptors have short cytoplasmic tails which interact with adapter molecules to facilitate signaling 39 Stimulates protein kinase 39 Cant produce a signal with a short tail they have to partner with an adapter molecule 39 Binds to aberrantly expressed proteins and sometimes to viral proteins things that shouldn t be therequot Activating receptors have immunoreceptor tyrosinbased activation motifs ITAMs 0 Not on activating receptors but they re on the adapter proteins 0 ITAMs are found in cytoplasmic tails Binds kinases Inhibitory receptors have immunoreceptor tyrosinbased inhibition motifs ITIMs 0 On inhibitory receptor 0 Engage molecules that block the signaling pathways of activating receptors Differences in NK cell response vs T cell response 0 NK cells kill target cells that express a combination of too little MHC and too many activating ligands 0 T cells kill target cells that present specific MHCpeptide complex for which they are specific The recognition even between the two is different but the actual form of killing is the same 0 Granzymes and perforin are used in killing for both Clinical consequences of a perforin deficiency 0 Failure to clear viral infections and a display of immunopathology mediated INFy and TNFa 0 In humans hemophagocytic lymphohistiocytosis FHL has been determined to be linked to mutations in perforin 0 Because of the lack of ability to kill you get to much expression of INFy Cytokines of the Innate Response Principle Source Biological Effects Interleukin1 IL1 Macrophages endothelial In ammation and epithelial cells Causes fever and synthesis of acutephase proteins in the liver Tumor Necrosis factor a Macrophages dendritic Activation of endothelial TNF 00 cells cells and neutrophils fever acute phase proteins and cachexia wasting of muscle fat cells Chemokines Macrophages epithelial Recruitment and cells fibroblasts activation of leukocytes IL 12 Macrophages dendritic Stimulates IFN y cells synthesis and increased cytosolic activity of NK cells Type 1 IFNs a and B Macrophages epithelial Induction of antiviral state cells fibroblasts and increases Class 1 MHC expression activation of NK cells IL 6 Macrophages epithelial Synthesis of acutephase cells proteins IFNy NK cells Macrophage activation
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