Micro For Health Prof
Micro For Health Prof BIOL 220
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5 Structure of HIV Virion contains a membrane envelope with a single viral protein m pr 39 7 Important in receptor recognition O Capsid made up of Gag protein 7 groupspeci c antigen 7 Icosahedral O Interior of capsid contains 3 enzymes 7 Pal reverse transcriptase 7 Prat protease 7 Int integrase Required for early stages ofretrovirus infection Maturational cleawge ofthese proteins occurs only after encaosrdanon and release ofvirion from infected cell FINIA twn Men nal strands Envalgpe phusphnlipid b ayemcen membrane 55 15 Egpm 1 gm f f erw game pra uct quot639 g Emerge Transcriptase pE N E I lintg rase p313 and f Pif fE EEE p1 39 pal gene pmdugtal f Matrix 3an Prmeins 39 gag game praducta PWH 3 E4 55 I Ths glymprmein fita ma CW receptnr n1 any cell that displays 3 cm IEEEPIDI 39 fir 39 39 l I ngII i i ll a 39 j Jl quot I Contains 2 identical RNA strands 7 7000 to 10000 bases long 7 RNA is capped and polyadenylated 7 2 identical strand function unknown I Speculated that it may buffer against a too rapid mutation mte due to inaccuracy of RT o Order of genes in Retrovirus R sequence 7 Present at both ends 20250 bases long 7 Important transcriptional signals that are only used in provirus U5 sequence untranslated 7 Does not encode protein 75200 bases long 7 Has cis acting regulatory signals PB Primer binding site 7 Specific tRNA primer for initiation of transcription binds Gagpr0tpolintenv 7 Gag encodes coat proteins 7 When gagprotpol in same reading frame they are expressed as one precursor protein that must be cleaved by protease 7 Gag expressed in much higher amounts than gagprotpol Q There is a variable length sequence following enV 7 Contains polypurine tract PP important in generating DNA from virion RNA 7 Untranslated sequence U 3 7 2quot copy of the R sequence Two different types of HIV 7 Mtrophic most important in initial infection I Macrophage is initial target I Individuals Who are homozygous for mutation in CCRS are extremely resistant but NOT immune to HIV infection I Individuals Who are heterozygous for mutation in CCRS are less susceptible to infection than normal people I HIV integrates into macrophage enome non dividing terminally differentiated cell therefore HIV cannot replicate Within macrophage I Macrophage acts as trojan horse in bringing HIV to lymphatic cells 7 Ttrophic forms made by mutation in m acrophage o CD4 with CXCR4 coreceptor chemokine receptor become infected with HIV in lymph nodes and other lymphatic tissues I T cells replicate Within these tissues to try to respond to HIV infection I HIV is continually being replicated in these T cells and as cells are killed there is massive stem cell proliferation to replace the lost cells 7 Increase in level of opportunistic infections begin as T cell levels drop to below ZOOcc FIGURE 21 Wral R plitatifj ih Humah JF I39W TW Ifin39jyTES SCEle li l 2th tmh In cmgm39pn m H h39 ihquotl39ec1ef h L mm M Til lj39r r39hrw1 jgth A sh 3amp2ng LEI hf ed with HIV 5quotar2rxir1g virus E39JElf TiEll ZES and E39 l ij afim Uh the CEW m Iiir haghihml FEDS timers B Enlargerth 0f 21 lzjml tirm JF the f I H39JvLJE IIEIM Ilih hell SM r F139e in All SHEEN39ng H IUIUFJIE virus l39har lhlhg huddhg hut hf hi3 Era3h 5m r1273 CE magni ed EUJJUD th TTEE has ea t h HIV THE jelly h IEEWE 1 thIE in the CUM hwmhr39ami iPhcgrf39iilgmph Tammi f Nagmhhml Frag 15 m H Eg mirt h In J I Fr39m39czch If if 339 quotEIquot 695193 rah Fa 71quot I jquot m HIV 1998 In 1996 deaths from AIDS declined for the first time in gt 10 years i Due to powerful therapies that retard the activity of HIV 1 AntiRT drugs protease inhibitors cocktails 00 Trend in industrial countries not representative of the world as a whole 39I39Expanding rapidly thousands of separate epidemics 00 Since early 1980s 40 million have contracted AIDS and 12 million have died 7 Nearly 6 million people 16000day acquired HIV 7 23 million have died from HIV disease including 460000 children More than 90 of HIV infected people live in developing countries but gt than 90 of the money spent for care and prevention is spent in industrial countries o HIV therapies cost N 10000year per person not available in developing countries o The region below the Sahara in Africa has gt 23 of the globe s HIV infected population and N 90 of all infected children 7 In areas of Botswana Swaziland and several provinces of South Africa one in four adults is infected 7 Life expectancy is falling in Africa 7 Unprotected heterosexual sex accounts for most of HIVs spread but also due to contaminated blood supply 25 of blood is NOT screened for HIV and this is administered to women and children India has 35 million HIV infected people o HIV is spreading into Thailand Burma and Vietnam and China o Epidemiologists have found that 7 Groups Whose human rights are least respected are most affected 7 As epidemics mature the epidemic shifts from the primary population to those who were socially marginalized or discriminated against before the epidemic began gender race economic status culture religion o In 1988 whites accounted for 60 of infections and blacks and hispanics 39 7 By 1996 38 of new cases diagnosed in Whites and 61 in blacks and Hispanics 7 Between 199596 AIDS declined 13 in Whites but not at all in blacks and Hispanics o Future 7 AIDS Will become more concentrated and expand faster in developing countries 7 HIV will enter areas Where it has not been seen be ore 7 Will slow in industrial nations for some populations but increase in marginalized groups 7 Cost of care Will rise dramatically 7 Highest priority must be given to finding a vaccine and making it available to those who need it most amp also to educate those people who need it most o Until a few years ago HIV infection was invariably a progressive lethal disease that robbed its Victims of dignity 7 Most medical interventions focused on treatments for pneum onias and other opportunistic infections rather than controlling HIV itself 7 Since 1995 advances have led to a shift in prospects for most patients who get treatment 0 Between 1996 and 1997 deaths from AIDS in the US declined by 44 Durin s me time hospitalizations due to IDS related complications opportunistic infections also dropped 7 Due to intensive cocktail therapies I Can this be maintained No long term data yet as to how long AIDS symptoms can be delayed with these therapies I Treatment is COSTLY I Some people do not respond well while others do very well 0 Ultimate goal is CURE not maintenance Management of AIDS is real but a CURE is probably not possible 7 Sexual contact 7 Direct exposure to contaminated blood 7 Mother to fetus vertical transmission through placenta and through milk 7 Contaminated needles 7 NOT TRANSMITED BY I SALIVA KISSING UTENSILS HUGGING SNEEZING MOSQUITO SWEAT o HIV infects primarily CD4 cells Helper T lymphocytes but can invade other cells macrophages dendritic cells 7 Replicates and is transmitted to other cells o Severity of infectious symptoms and expected life span after infection depends on infectious viral load o At start of infection there is a high level of viral replication as measured by viral levels in the blood and by a drop of CD4 cells CD4 cells normally are about 800 cells per cubic millimeter of blood True AIDS occurs when these cells drop below 200 About 3 weeks into infection acute phase many people display symptoms like mononucleosis 7 Fever enlarged lymph nodes rash muscle aches and pains and headaches 7 These symptoms resolve in 7 3 weeks as immune response begins to control infectious process killing of virally infected CD4 cells by CD8 cytotoxic cells 7 Antibody molecules produced V 4 9 a a1 axiuf 39 g amp xii 3111 a 4 LA d hm 14Jg hstt i E D u T Veg 4 11 L44 wmia Lwr nai W3 SOURCE n u A H aucr WVEEKS EIGL H1 Ama s wa I temal Med r M zine VG 39i I24399 YEARS 6 a 3 8 9 m H u 1 I 1 n a SEE Suzy EE gag Qwimm Ex E p EH3 9n 3 Ema CHROME PHASE o By siX months rate of Viral replication is at a lower but steady state 7 Seroconversion antibody levels detectable and can be measured by ELISA at 6 months 7 Level of virus replication is patient dependent and will determine the subsequent rate of disease progression I Generally 810 years pass before major HIVrelated symptoms appear I Chronicprolonged phase of infection 7 Over time CD4 levels gradually fall When less than 200 cellscc patient has AIDS HIV is able to infect cells other than CD4 7 In addition to the CD4 antigen there are other coreceptors to which the HIV binds I CCRS found on macrophages I CXCR4 found on T cells 7 If these coreceptors are mutated on the cells then HIV is unable to adsorb attach and the individual is resistant to infection HIV replicates only when the infected cell replicates 7 When specific immune response begins specific helper cells die o HIV mutates at high rate and immune response cannot keep up with antigenic changes after 1012 years Q As CD4 levels drop below 100 HIV levels in blood increase 7 Bacteria that are normally contained begin to proliferate and cause opportunistic infections I Pneumocystis carinii and toxoplasmosis 7 Once these symptoms begin to appear the patient usually has 12 years before the disease is lethal O Once HIV replication begins the RT begins to replicate both the viral genome and the host cell genome host cell activation and replication 7 Protease enzyme cuts new viral proteins into forms that are packaged with the viral RNA two identical copies of RNA 7 Viruses bud from cell pick up host cell membrane and infect other cells 7 When these cells become specifically activated the virus replicates and cycle continues with many mutations 7 Most mutations probably make non productive virus while others give resistance and change antigenicity of virus O AntiRetroviral Drugs 7 Block viral replication in two ways Inhibition of reverse transcriptase prevent integration into host genome by preventing RNA DNA transcription iNucleoside analogues resemble natural nucleotides but prevent completion on growing strand AZT 1987 zidovudine Inhibition of HIV proteases block catalytic site of HIV protease preventing it from cleaving newly made HIV proteins o Early beliefs of HIV disease 7 Suggested that only a few CD4 cells were actually infected and that HIV replicated weakly for a long period of time I This view implied that most ofthe lost T cells were killed by a mechanism other than by HIV replication if this were true then HIV directed dmgs might not be able to prevent this T cell killing 7 It is now believed that HIV replicates fast from the start I HIV levels remain stable for several years because body responds by making very high levels of CD4 T cells replaces those that are killed o The strength of the initial immune response has a significant effect on progression to AIDS 7 Those who respond with strong CD8 activity get greater suppression of viral replication early in infection and progress more slowly towards AIDS than those who mount a weak response 7 A strong initial response helps to later manufacture the subset of CD4 cells that specifically react to HIV I Once these speci c T cells are lost they may not come back with treatment even though other CD4 T cells are made to increase T cell number to greater than 200 At any stage viral levels correlate with prognosis 7 Patients Whose viral levels fall into the undetectable range and stay there are most likely to avoid progression to AIDS Thus the amount of virus in the system plays a major role in determining a patient s eventual outcome 7 Therapy aims to shut down viral replication 7 For those patient s Whose immune systems are suppressed this is best way to keep viral levels down 7 All patients must stay on medications I a arhuddinzg mammal 15 msipn 1 i i I f P hfa39ti iquot quot i quot I quot 39 39 assembiy a 39 r lt I I if 39 5 i Ia W Ifquot 39 I quotA I r V V nquot u quot r v 39 new m i 39 39 i 1 vquot u 1quot 39 57quot I 333133 i A 391 V 455 V x i I 73 y 332 3 I 3quotiii wzcsm I quot9525 i lt 1433 i m i 3 51quot 39 39I quot39I39 quot 39 iquot quot1 W 39 gt 1 s 13 wq u 4 iii922quot5 393 i 21 514 75 i a 39 w egg3 34 1v 39 V a39 v i i 4 i a 39r39 ltr 39 quotw 39iPi ii 39iquot 153 4 a i V 7 39 r 39 39r i quot39 In 251 r 3 iv nun 1 i tkgug f zgz39s39jr 139 Vii J u 7 V T a b 7 V i 7 I i i a I EUHI EE tranacrimase 7 and FREE 1 i i 1 i i 39 LTFi LTFI i l i 39 quot 7 a i i quot39 1772 397quot 397 A i Pf g n i RNA Win Nuclear transl cati n integrase Ti z i i i i ii i E Jmegratiaun 7 DfCDNMnm V gamma LI If segagl igw meral DNA 3 Humans K39 Cyt p iasm 39 quot quotFiatEggs mediates 39 FEMSe by 39 quotbUdding Em pmiein 1W alters plasma membrane Gmlgi Call genm iii DNA apparatus 1 i Transiatiun if i viral gprmein iii re iiiitiii iyii i i typical riiiiirm Aimptiin iii pinitriiiii iy Tifipiir39miiiiiiii mimiriii in ii iii in partial inmatng iftii viii mpii iii ginirmian ifiDNA E E quotHE2 frag quot h M 1quot E E t w E 23 E r Tquot 39 H 2 m quot l Pea E A r 39Fai 44 27quot EL E 395 E a arma E I 2 2 E m m M a an Mr H 324 a mi Livi Ea ofinwmaiim requii iii division in iii iiiiin ii not required ii iiiiir transport if M iii integratiipmvim M ii iii own ii tilt is W riiiii om iii iimipmmriii iiiim i ii ii iii immcriiiian rennin u g6 x i ii ii iii ii ii iii i iii pituiin iii iii 39 o Optimal Therapy HAART ighly ctive nti etroviral herapy 7 Consists of triple therapy I Two nucleoside analogues and a protease inhibitors 7 Dosage ofpi11sday empty vs full stomach side etfects etc may be too much for some patients andthey stop therapy 7 Least number ofpills is 8day and most is 24day I Cost N 1000012000year I ntl rugs DW 0111 t e Mari 6f quot quot 7v n c i9 rrvquot I 1quot a r quot 341 quotquot quot 44 9quot39 quot Reverse Transcriptase lnhbtnrs Nuclesude Analogue 39nsinewamdn litpillalies ay39 I V an emptyst Emach Lamivudine Epivir STE 1 pm 2 times a clay sravudine Zerit am 4 Zalcitabihe iHlVlD dde l pill 2 times a day 39139 pill3 times 3day ZidevudineRetro tAZT39 1 pill 2 timesa day Pill centainin39g lamivudine l pill2 times a day arid zid vrudine Cumhiw I Reverse Transcriptase Inhibitors Nonnucleoside Analogues Delaeircline Restripterl Nevirapine Viramune r i p ili 2 times aday a j i Pr uteease Inhibitors Indinavir Crixivanl 2 ins3 tithes a day 3 plil39s3timesfa clay 39 Neifinavir Wireteptl 39 1 with some feed Ritenairir North within 2 heurs ef didannsin39e SaqUinavir ilnvira39se a hardgel 6 pills 3 times a day er 2 pills2 times39a day if taken with ritenavir with alarge meal ca psule Fertovase a 39 suitgel capsulel tea of Aprii 1993 3quot 25quotquot 3quotquot quot5 Jquot 39quot quot in a 7 r V 1 if i 3 iiiamp iiitiE urn at an Verifier and 3 while 04 at 32mm ate if gt39lf39f39 rr 1JM 1 quotquot3939Fd1gt39n s quot 39 I we in I 1 F quot 431 a r r 1 5 39u Viquot a sL39ir LiLut izktrsh LLBu a J v 1LCQLUSELI atrtda gum 42 halt a quotail 139 1r W 39 w 14 z i11quotjquotquot H1 quot Lia s r 5 139quot nuquot 5 17 I quotF F r an 391 3153 if inlzln3ril 13l L611 Li I iiltinLlniizq mini LLHII E CJ a w r yquot I i 1 r t T I 395 h 39v 5 w w trin it itnai i at 11 my gainh 1 iTE39JLtir lf t i rtstg t g n a 11 1 r n gut a 3de 3371 at tuit i vi t nil ting v i zisettur 53113 11 quotWTquot quot r x ri n A f 1 flu I a quot 3 1 1 ii39 iaediiiit e A 11123 R U iu a39 mu Wreath Silt UI ELH LJLLL J PM 4 pills 3 times a day mixed ihte water Within an heur of antacids er didanosine 39 5 pills 2 timesa day or 4 pills 2 tii i lES a dag39 if taken with saquinavir with feed and net i I v b 39 39 39 39 39 V t a v I t v J 39n39ewr pathli 39 Nailsea ciia rrhea pa nc39reatitin ammatie n peripheral USualIy39 nehe I 39 t Peripheraineumpathr Peripheral neumpathy meuth in ammatien pancreaticin ammaton Nanseaheadacheanemia neatrepenia redacted levels of rieutrephil white bleed EEIISLWEBRHESE insomnia 39 39 Same as far z idawdine het iFi i heetice heeatitis A Rashhepatitis Kidney stenes nausea hea d39ache blurred Visit n 39 39 an em pty stemath or with a gianti fat snackl Famine55 rash metallic taste in Mouth ab39nermai and not within Zheurs of didanesine distribLitiehef fatelevatedtriglyceride and cholesterei levelsglucesein39ttileran39ce39 39 Diarrheaabhermal distributien of fat elatiated triglyceride andchelesterel levels glucese intolerance Nausea verniting diarrhea abdominal painheacl ache prickli ng 39sehsati n in skirt hepatitis weaknees 39 f 39 abnormal distribution of fat elevated trig lyCeride and gchelesterel levelegluceee ihteleranre 39 39 39 Nausea diarrhea headache abhermal distribution ef T fat elevated triglyceride and Charlastem levels glucase inteierahce SUM FILMS in x 39Iquot 11 IL 4 4 i a 1n nn quot1quot r a w 39 a a 39 393 1 u39 quot Iraf quot39 nh x39 P g a L tLLE E quotMi 1 N run Em ma1 1111 AL ii 3153 hisas it Deity t t i 1 quot 391 d r en v 4 iu u v Ian war 5 1 gt are n I 4 13939 39 39 EELHHQ m 51 1quotEQ3CLL3JR1LS it 3511614 titan ailTM LZL Ht itg t I l I I a a h v i 1 i r I v V swag 1 4H J8 quots gr Fae3 4 x 1 39q 515 5 7 i at l 1 Mali zue a HE LeiM quotw ygiayx till UH3 11352125 1111 395 MEL195 1 11quot Sit3 1 4 39g 1 e rw sWyx quoti aslp 1 I I hi quot1 1 3 1 i 1 in J x x 1 1 Chi332 5L EJ t5 that M a 39 tum LIELe JEL ELSLM Eizah Luff Hui hi u l u uh 3 1quot I 1quot quotg i39v39uquot 391r r39 3quot quotI r s il In 1 39 1 Equot 39quot quot 139 quot P 39t 39i urn311 3 hi mum 1213 hi E n 1 4113 t um ing139 d1 metre I u 31quot an g I vrr 39 quotTit391 E H Ia Hi 4 quot14 Iquot I 1 39 JCI 52 th Em Hulaa mm at rue it an Ev 111th M ht45 l wnm WHGHD n am mDH A HA man Dm m M Auwm n m 347 quot lAa 911 u 1 u 3 13 IA 3 A u A A llnlrlhllLrIAA kw w 1 A Hiawig 1 A1 A AA A mm m m inqqu mm A Amm inw A A A EJUEUH A A A W W A A A M A w a H H H AM AA w H M H 3119 A A H rmam immlm H A w mm mm gmnw mwmAA A mm minw H H M EZEEAQU A A M H H A AA y A m mwwm m mjm W A A H A 5353 A AA A w A A A H A u a 1 M l f A y a L N u A u x t A I u a t A A 4 I u Au mgwi mmam A A A A mm w mm quwjmnnv mm H AA mimm A M M Eggnog H H J m A A A H x r A V I f r u 42 L A 141 a lfm nm 4 x I 1 12 Al I 1L L r SJ A A A Lu k9 qu v Av 121 A P A A r K g iquot A 5 1 J ansr l39 A A ENE an H H A A w m mme Hidxm m m AA AA Am A A W AA p A A A w H g A A A u a I m A A w A A w u A A A a x A A Aquot a v x 1 v x r r L A A A n a a A x A h A A 1 A A 4 AA AA a v A z A r A A A s V A u A 4 J urnl Aanmu iq A A A M 1 IV A A A A A A L l mi A A A A A H A s m A A y I A A A F A a n 1 1 A y u v 7 391 t I V s AA v A n A n u L A A I a A m Drug Resistance o Mediated by mutations 7 Nucleoside analogue resistance may be caused by a single mutation to reactive protein 7 Protease resistance to drugs usually requires at least two mutations in a single gene o HIV makes N 10 billion replicatesday 7 Done Without accuracy genome of each new particle probably differs from parent genome in at least one spot 7 Thus every mutation able to contribute to drug resistance is likely to be made in some of the particles TOR EXIST MG INHHB EASE HW FRET U E SITE ACT DRUG BINDlNG f AREAS A ALTER NHIEATQR DER U N Thus if patient has never been treated any compound that is given will encounter some HIV variant that is already resistant If antiretroviral drug taken drug resistance can be attained by only lt5 mutations in a genome will block not variants but resistant forms will proliferate and some of the semi resistant variables will continue to divide and allowed to generate other mutations towards resistance 7 Antiretroviral drugs will select for variables mutants 7 Use of polytherapy vs monotherapy lfvirus is detected in blood after 46 months of therapy then probably a variant that is drug resistant Must alter therapy depending upon the resistance type found Viral levels assessed by viralload assays which count copies of HIV RNA in a milliliter of plasma The number of viral particles is 2 the RNA count 7 Current test sensitive to RNA concentrations of 500 or more copiesml 7 Within 1st 8 weeks of therapy viral loads should drop about 10X by 6 months undetectable 7 Triple therapy successful in 7585 of patients Viral load tests measure viral RNle7 Number of Viral particles is 12 the number of RNAs found why 7 Each viral particle contains two RNA strands In 1996 researchers examined l600 samples of blood from untreated patients and compared viral load with prognosis of survival 7 Found HIV replicates at high rate from start 7 Found viral load directly correlated with survival I 70 with gt30000 copiesml died Within 6 years I Iflt500 lt1 died in 6 years average gt 10 years PATENTS SURWWNG PERCENT 40 20 WNITIALWHAL LOAD HW RNA Epies per miMHiter of pigma a L 77157WJ gt H7 9 I A h 39k V I M V I I 7 V gt V I t ltii wwwwwvmeMwwwwwwwww f J 500 501 TO TO mm 1 001 T MING 2 30000 2 J W Emg AFTER WRAL LOA i 6 WAS M EASURED o New challenge now is to identify cells that contain provirus resting T cells in which Virus does not replicate 7 Current drugs do not do this 7 To develop these types of drugs researchers must develop new viral load assays that measure infected cells I This Will 2110 for measuremenw of success ofnew treatments to eliminate proviral c ls I these cells would have no markers speci c for Virus how would you nd these and identify them Trial patients versus field Clinic patients have different results 7 Trial patients have 7585 success 7 Field patients exhibit 50 success I Field patients more heterogenous I Field patients often start later in disease process I Field patients often do not follow protocol andor stop when they feel sick I Field patients may have been on antiretrovirals previously and contain resistant forms Even successful therapy does not restore immune function totally 7 Mix of CD4 cells may be abnormal may not recognize as many pathogens or may be less effective Resting T cells with provirus are not killed by current therapies 7 If become activated will replicate virus 7 Certain cells act as reservoirs of HIV I macrophageshort lived brain neuronslong lived How to activate immune system 7 Immune cells must see antigen to destroy cell I Give substance that activates infected cells 7 Give HAART to prevent other cells from being infected I Remove stem cells grow in vitro and add back I Remove stem cells add a gene protective against HIV grow in vitro to expand and return to patient I IL2 therapy only cells with IL2R T and B cells o Other Strategies for preventing infection 7 Block integrase enzyme 7 Knock out Zn from protein that needs it in order to draw HIV RNA into new particles being made in cell 7 Use of antisense DNA to inactivate two tar and rev that are necessary for manufacture of other proteins 7 Block entry into cells I Must bind to CD4 and coreceptor proteins I CD4 blockage not very effective but blockage of other coreceptors effective SA Sept 1997 7 Use altered designer virus that infects HIV infected cells and kills those cells HIV Vaccines Natural immune response that vaccine elicits does not destroy HIV in cells 7 Will serve to block HIV from infecting new cells humoral arm of immune system No vaccine made yet to activate cellular arm 7 Will act as a protective mechanism for infection 7 Cannot immunize with vaccine against all variants of HIV anger of using both Whole killed viral vaccine and live attenuated viral vacc39ne O Best vaccine Will activate both humoral and cellular branches of immune response HIV env protein is a gpl60 7 Composed of gp 120 that interacts with CD4 protein on TH ce s 7 Composed of gp 41 anchors gplZO to membrane of HIV Antibodies to both gpl60 and gplZO elicit Ab formation and block HIV infection in test tubes 7 Only recognize strains from which they have been made not other strains I Antigenic sites may be different between lab strains and patient str 39 I Difference in coiling may expose hidden antigenic sites that are not protective in patient strains o In vivo strains may have surface antigens that are highly coated with sugars that would serve to block the ability to recognize antigenic sites o People who are infected with HIV but remain healthy make a very small amount of antibody which can neutralize viruses from many different patient isolates 7 If understand differences in antibodies made then might be able to develop vaccine for HIV Making a killed virus vaccine requires a rigorous inactivation procedure 7 Residual genetic material may be dangerous 7 Harsh treatment makes vaccine less effective 7 Inactivation may cause HIV to lose its gp120 and make it less effective Envelope proteins embedded in pseudovirions empty lipid shells To make T cytotoxic response must associate Ag with cell membranes and self MHC 7 Viral gene inserted into host target cells by viral vector ENWROTEIN If Fm LABORMBHV 9quot GROWN Hw H z I V 39 w 39L EDHATCJH F PATIENT RQWN HW ANTIBnm I mm n quot quot quot at Niar wgv r h a K M39FI39lw 3quot may 39 Aw 3quot ur l quot elp 39 39 39 39 1m quot39 39 M r 77quot 39 r m Mnc wmwr V v Mnm pm v quot uq m H w quotm m wquotquot lu m Jru M410 39 I 5 u k uh I ww P mu H a wer c f H in pul 39 39 g39 m Hunquot q 2 wkquot id 39 i p99quot i gun 1 amid Hmsaw39 H vquot Mk 39 I v 9 V II a 39 Mark quotWquot Hquot 3953 ll wiquot quot Eryn 5 I ndquot 39 39 39HWW ll1 H J v gw39m f I 39 quot311 b a39ay v 62iquotfm iquot a 39139 I I V39 v x JJ I I A er 1quot W39 I J I r aly39l39l uy39lldquot l 1121 JI r II I N nun39uwlsr li39li H L rquot y i K r1 391 039 J quot fr v39v l I fquot 1 hr v r H H V J I w 5339 if quot Illii H I 3 3 41 139 Iquot EV ixuv v I39a 39 I ii I q x 34 339 ll q l JariLi H 1 M1 i vuil i l Jaw 39 39uL39HV 9 39LI 3 39 l E v 1 z l39 I HIM E j n w 39 lf39x I I Most common Viral vector being used currently is the Canarypox Virus 7 Canarypox is defective and will not multiply 7 Genes for env and gp120 and other nonsurface HIV proteins inserted into canarypox gag and protease genes 7 Have proved safe in human testing and have elicited a modest TC immune response I Need to have Viruses that cause production of larger amounw of HIV proteins 7 Multiple shots boosters o Other researchers use of Viral peptides 7 Does not elicit good immune response I May be degmded too fast n g 23 39l V W 1165 1 mac in fl CANARYPDX 39ak 4h w FD 39 r Lia Nun Mm m 3 U j 1 I CELL CELLS PHIMED TO ATTACK HIVINFECTED EEEDMES Injection of naked HIV DNA 7 Does get into cells and direct production of viral proteins I Does seem to cause production och I Must evaluate safety and effectiveness o Combination vaccines 7 To activate both arms of immune response I First may be exposed to canarypox virus carrying env to stimulate cellular response I Months later receive pure gp120 to elicit antibody Rx 7 Need to improve delivery system and strength of response use more HIV proteins quot A4 1quot Jv4quot l t r 39 l quotquotquot lI39 r V quot 39 vr uvw r n 39 n v a41vntvo HW4V 5 nr dunr Wh n 4 quotquotamprrgt quotquot lquot39 1 v quot 395 39Iquotquot 1 lquotquotquot ilvz 11 quot39L39L39lz Iiquotl quot39 39 flquotw vH39 quotl quotI quotl l U quot quotH n Hup U u A Hun quotLa 39 M 39l H 4 r 1 H u n a quot39r u 39 vl A v gt y I V n14 r 39quot Ovv 39 lquot I I r ll a n394 l M Iquot vIl V J qu a iv INA 2 H u I w 39n V uu A39quot Iquot Iquot H l l9quotquot 39 39 quotquot 7 I l Iquot l 1 quot V 1x 9 1 4 u um I Hr quot mu A quot quot llla39tll1i nnstltuan3ts quot I39 39 I l 39I a r quot 5 t Ia quot In R 39 J wquot 39 39 v 39 n gt7 r x r n 39 I l quot v Iquot 39 HIJ rr u AH um H quot quot v39u 39 vllquotl H 1 Ar v u gt Vaccines Eliciting AntiHm Antibadies 39 39 Viral surface pruning InlphasElandflltriangQ j safea ndgimpletnprepare V vaginaIlcltedantllmd i have su chasgplZ WhlchfexaminesafetyV I 1 I 39 v falled tinrecclgnlzel5lllfmmpatler s I k 3919 quot2quot W 39 N lu dersmdy 5 3hauldp39re5ant Htwrsurfacev SillghtrlskthatPreparati hsmight I inhuman I w p fipl t ih lln a1ir latljvelynatural include samei actiyeuiruslnactlvated gt I 7 quot 1jmnf rm ti Elmleiquot virLls mightvsh dits pratelnsan 39 39 r39 39 39 v g W I 39 g I j l I Imprepare a T I becameine ectlve I 39 Psg dwirlnns Cl seta Phaseltrl39als 39 PreseljfthllJsprf t pm felnai 1 gEll iEUlttd l dLlCEal ldtDEl lEurE quot a i cialvirusesl f finf ralatlvelywnyamla l g r 1 lgng ltarm stability 39 39 1 quot l39vkmnfarmatlunqg 39 39 39 39 i 39 3 Vaccines Eliciting Cellular Responses f Liveyyewctaru39rirus r39 I I ll phaslelltrlal39s39 V H I I Hawkscammmlallmum 39C dmpllcat d Id prenar urrent 39 lfnrl39lHl llr Viruses ll and kindsafarlralgpmt elns 39 vactlneselititm des timmune englneeredmcarrygenes39 39 39 g 39 A f i z plgduCEdw V 39 response I annealinglllemtelnsl 39 39 j f A 39 39 I I Naked Drum j lnphaselt39riia ls f I 5 39 jSimpleandlnegtpensiue 1 39 39 7 Slumewnrrythatintegratinn quW39 cantai i ng FlE or mare I I j 39 39 39 mnremlre 39 39 I c T r 39g39El lESi t human cells uldf harm i Hl39llgenes w A quot I patients I 39 HIV peptides I I39 I f In pha eltr9lals7 quot quot Stimpl ttl l j n f EilluzltstmnglmmunEIIequnse f 1 letelnfragments 39 39739 gt M 39 1 39 39 V 39 v 39 Vaccines Ellciting Antibody and Cellula rResponses l i l 7 m 39 I quot th39ls Ell t 39PIEparE l elements h as pureA 7 7 quot 7 l SIquot thel l l llTILl Er EiSpsj SEl 39 f 39I gpl2 pmteinplus y 39 quot F HWME rcanan j xvectar 39 39 39 39 i 39 LivE attenuated Haw Nut under Study ini39 5 1Masticlaselymimi s39leilway Virus could potentially cause disease V 39 hrum l lslib ngj assessed interfere with ll lf Efti US HW39S 7 39 in nunhuman primates 39 ability It replicate 5 IM MIME HIV VIMmes Pmspelcrs and Challenges SCIENTIFIC AMERICAN Julir 1998 21 Use of live attenuated vaccine 7 Must delete genes critical for HIV replication While maintaining antigenicity of virus 7 Recently a group of physicians volunteered to be given an attenuated virus so that the response process could be monitored I Volunteers feel that value of testing this approach outweighs the potential risks to their health Attenuated SIV in monkeys I Some have proved very effective at inhibiting disease progress ofviiulent strains ofSIV I Protected monkeys do NOT have high Ab s or Tc cell activities so basis of immunity unknown 7 May be combination ofAb Th and Tc At present there is no proof that vaccines will provide longterm full immunity to disease and may even lead to disease Vaccines may give body a head start for protection and thus lower initial level of viral load Wide genetic variability of HIV will limit effectiveness of vaccine I HIV from one patient or one part of the World would be different that from another 39o Vaccine use will not be available for at least 5 more years 7 Phase III trials necessary 151 V39Dr39ugs fundeir studywo uld blackbindi ha i w W PROTEIN IINITEGRASE REVERE I TRANSCRIPTASE inhibi transcn I traverse ptase IIIIIISII Hiya d i mm 3 lat U m MI INFECTED CELL Cu 39H finquot JMJ II b Inhibit protease Some existing drugs SCIENTIFIC AMERICAN July 1998