The Impact Of HIV-1Specific Cellular Immune Responses In HIV-1-monoinfected And HIV-1/HCV-coinfected Individuals

The clinical therapy and the relevant pathogenic mechanisms of humanimmunodeficiency virus type1(HIV-1) and hepatitis C virus (HCV) coinfection areChallenges facing the clinical and research focus. HIV-1and HCV share the sameroute of transmission, for example, blood and blood products, sex transmission,mather to child transmission, so the phenomenon of HIV-1/HCV-coinfection isvery common. As the highly active Anti-retroviral therapy (HAART) has beenextensive and standardized applicated, the morbidity and mortality of opportunisticinfections have significantly decreased, and liver damage cased by co infected withHCV has became one of the most primary reasons of death in AIDS patients. The lowefficacy of anti-HCV, the increasing of adverse reactions in HAART and thegenotypes and quasispecies are complex epidemical in China are hard to treat, thesemake the therapy of HIV-1/HCV-coinfection more difficult. The reason is not veryclear until now, so it is very necessary to study in depth from the aspects of virologyand immunology to HIV-1/HCV-coinfection.Many studies have been confirmed that the cytokines play an important role inthe process of disease progression in HIV-1infection, and the studies also showed thatthe IL-4can restrain the level of virus in vivo,IL-7plays an important role inmaintaining the survival of lymphocytes,IL-17may take part in the immuneresponse,IL-21can stimulate the cell proliferation and activity, IFN-γis the mostimportant cytokines of the immune system in vivo for resisting to virus infection.We speculate that the above mentioned cytokines in HIV-1infectors would effect by coinfection with HCV, but the effect between HIV-1/HCV-coinfection and thecytokines has not been elaborated much, so it is necessary to study the secretion statusof the above cytokines in HIV-1/HCV-coinfected patients. The studies about specificcellular immune response in HIV-1infection and HCV infection all have promptedthat it is important for the specific response of CTL to the control in HIV-1infectionand HCV infection; there are a few studies about specific cellular immune response ofHIV-1/HCV co infection reported. Because of the differences in geographic andethnic, it is necessary to study the specific cellular immune response of HIV-1/HCV-co infection in Southern China. We detected the immune response of CD8+T toHIV-1synthetic epitope in HIV-1infection by the method of ELISPOT in theprophase studies and faund that: the frequency of IFN-γ secreting cells induced byHIV-1Gag peptides is the highest. So we selected the peptide library, whichcomposed by12overlapping peptides compound by the amino acids in the P24region(Gag133~191) of HIV-1, to be specific peptide epitope,and began the study about thespecific CTL response of HIV-1in HIV-1/HCV-coinfection and HCV mono-infection.Part Ⅰ To detect IL-7,IL-21, IFN-gamma mRNA levels of CD4+Tcells, CD8+T cells and PBMC in HIV-1-monoinfected and HIV-1/HCV-coinfectedindividualsSubjects: The object of our study was25HIV-1mono-infected patients,22HIV-1/HCV co-infectors which were all treated by HAART above12months and20healthy person in Southern area of China.Methods: We cultivate the CD4+T lymphocytes, CD8+T lymphocytes（whichselected by PBMCs through the technology of immunomagnetic microbeadsseparation）and PBMCs of above mentioned three groups with or without a peptidepool containing12overlapping peptides in HIV-1P24. And then we tested the mRNAlevels of IL-7, IL-21and IFNG secreted by CD4+T lymphocytes, CD8+T lymphocytesand PBMCs respectively, through the technology of RT-PCR. And we analyzed resultswith one-Way ANOVA.Results: Comparion of the mRNA levels of IFN-gamma, IL-7and IL-21among 24HIV-1mono-infected cases with HAART、25HIV-1/HCV Co-infected cases withHAART and20healthy control casesAfter cell culture with P24peptides stimulation, the IFN-gamma mRNA levelsof CD8+T cells in HIV-1mono-infection was significantly higher than that of CD4+Tcells (p<0.05);After cell culture with or without P24peptides stimulation, theIFN-gamma mRNA level of healthy control was significantly higher than that of theother two groups (p<0.05); After cell culture with P24peptides stimulation, theIL-7mRNA level of HIV-1/HCV coinfected group was significantly higher than thatof HIV-1mono-infected group (p<0.05); After cell culture with or without P24peptides stimulation, the IL-21mRNA level of three groups was not significantlydifference (p>0.05).Part Ⅱ To detect the secretion of IL-4, IL-17A, IL-21,IL-21R, IFN-gamma ofCD3+T-lymphocyte in HIV-1-monoinfected and HIV-1/HCV-coinfectedindividuals by ICSSubjects: The object of our study was24HIV-1mono-infected patients,21HIV-1/HCV co-infected patients which were untreated with HAART and20healthy person as healthy control in Southern China.Methods: We detect the levels of IL-4、IL-17A、IL-21、IL-21R and IFNG in theCD3+T lymphocytes through the technology of intra-cellular CK staining(ICS) aftercell culture with or without P24peptieds stimulation. And we analyzed resultswith one-Way ANOVA.Results: Comparion of the T lymphocyte proportion of IL-4,IL-17A,IFN-gamma, IL-21,IL-21R among24HIV-1mono-infected cases without HAART、21HIV-1/HCV Co-infected cases without HAART and20healthy control casesAfter cell culture with P24peptides stimulation, the T lymphocyte proportion ofIL-4,IL-17A,IFN-gamma,IL-21,IL-21R were not not significantly difference with thatafter cell culture without P24peptides stimulation (p>0.05). After cell culture withoutP24peptides stimulation, CD3+IL-4+T cell/CD3+T cell of HIV-1mono-infectedgroup was significantly higher than that of healthy control (p<0.05); CD3+IL-4+T cell/CD3+T cell of HIV-1/HCV co-infected group was not significantly higher than that of healthy control (p<0.05). After cell culture with P24peptides stimulation,CD3+IL-4+T cell/CD3+T cell of three groups was not significantly difference(p>0.05). Whatever cell culture with or without P24peptides stimulation,CD3+IL-17A+T cell/CD3+T cell of HIV-1mono-infected group> that ofHIV-1/HCV co-infected group> that of healthy control, that of HIV-1mono-infectedgroup was significantly higher than that of the other two groups （p<0.05）;CD3+IFN-gamma+T cell/CD3+T cell of HIV-1mono-infected group> that ofHIV-1/HCV co-infected group> that of healthy control, there was not significantlydifference among that of three groups (p>0.05); there was not significantly differenceamong CD3+IL-21+T cell/CD3+T cell of three groups (p>0.05); CD3+IL-21R+T cell/CD3+T cell of HIV-1mono-infected group> that of healthy control> that ofHIV-1/HCV co-infected group, that of HIV-1mono-infected group wassignificantly higher than that of the other two groups （p<0.05）.Part Ⅲ To detect HIV-1specific cellular immuneresponse inHIV-1-monoinfected and HIV-1/HCV-coinfected individuals by ELISPOTSubjects: The object of our study was the object of the first part and the secondpart except healthy control.Methods: HIV-1P24region overlapping peptide pools as the specific peptideepitopes. HIV-1specific CTL response frequency of PBMC was detected by themethod of enzyme-linked immunosorbent spot (ELISPOT). And we analyzed resultswith statistical methods. Analysis HCV co-infection influence on HIV-1infectioncell immunity from different angles,and HAART influence on HIV-1infection cellimmunity.Results: Comparion of the specific CTL response frequency of HIV-1mono-infected group and HIV-1/HCV co-infected groupThe HIV-1specific CTL response frequency of HIV-1mono-infected group withHAART(13/25) was significantly higher than that of HIV-1/HCV co-infected groupwith HAART(5/22)(P<0.05); The HIV-1specific CTL response frequency of HIV-1mono-infected group without HAART(8/22) was not significantly higher than that ofHIV-1/HCV co-infected group without HAART(6/21)(P>0.05); The HIV-1specific CTL response frequency of HIV-1mono-infected group with HAART(12) was notsignificantly higher than that of HIV-1mono-infected group without HAART(p>0.05); The HIV-1specific CTL response frequency of HIV-1/HCV co-infectedgroup with HAART was not significantly difference comparion with of HIV-1/HCVco-infected group without HAART (p>0.05).Conclusions:1. HCV co-infection may reduce HIV-1specific CTL response of HIV-1infection,this may be one of HIV-1/HCV coinfected HAART poor efficacy reasons.2. Cytokines (IL-7, IL-17,1L-21R) play an important role in the cellular immunityof HIV-1infection. The cellular immunity of HIV-1infection can also beinfluenced by Th17and Tfh pathways, HCV co-infection can influence thesepathways above mentioned. IL-7plays an important role in the cellular immunityof HIV-1/HCV co-infection.