The Study Of Safety And Efficacy Of Adoptive Cellular Therapy With HLA-A02 Restricted HIV-Specific CD8+T Cell

Objective: To select HLA-A02 restricted conservative HIV antigen epitopes,develop method to manufacture HLA-A02 restricted HIV-specific CD8+T cell product for clinical use.Recruit cohort for the adoptive infusion of the cell product and evaluated the safety and efficacy of the therapy.Method: 1.Subjects: 1)Epitopes selection study: cohort participants were individuals infected HIV via contaminated commercial blood donation in 1994-1995 in Shuangmiao Village,Henan province.This cohort is characterized by similar infection timepoint,same transmission route and close genetic background.2)Safety and efficacy of adoptive infusion study: cohort participants were MSM HIV+ individuals commenced c ART in acute infection in Beijing You’An Hospital.This cohort is characterized by same transmission route and early initiation of c ART.2.Experimental Methods: Blood samples were collected from participants and PBMCs were separated to: 1)In epitopes selection cohort: DNA extracted from PBMC to analyze the HLA alleles and HIV proviron DNA of the participants by PCR to investigate the correlation between HLA-A02 and virus mutation.Reactions of the epitopes by the PBMC were tested by Elispot.2)In adoptive infusion cohort: co-cultivation of the PBMC and epitopes peptide in non-serum environment with cytokine,analyze the count and phenotype of the cells when cultivation is finished.Pyrogens and other contaminations were tested and products were packed when suitable for adoptive infusion.Participants received the infusion and followed-up by 5 time points.In each time point,symptoms,vital signs,laboratory tests were collected and acquired blood samples to separate PBMC for further fluroscence cytometry tests.Result: 1.3 epitopes were found HLA-A02 restricted(pol-IV9,gag-SL9,pol-YI9)and 5 epitopes were found with better resistance to immune pressure induce mutation(pol-IV9,gag-FK10,pol-AM9,pol-VL9,nef-PL10).Breath of recognition(>1 epitope)by immune system could resist epitope mutation better.2.The cell product manufacture method was able to expand CD3+T cells,CD3+CD8+IFN-γ+ cells,HIV would not replicate through the procedure and products were suitable for clinical use.3.The therapy was safe with a low rate of adverse effects which were controllable,there was no occurrence of severe adverse effects.4.The therapy could down-regulated the membrane expression of PD-1 and CD95/Fas of the CD8+T cell,proving the efficacy of the therapy.Conclusion: 1.Some of the HLA-A02 restricted HIV antigen epitopes(pol-IV9,gag-FK10,pol-AM9,pol-VL9 and nef-PL10)are better resisting mutation from immune pressure.2.Wider breath(>1 epitopes)of immune recognition of the antigen epitopes can effectively resist epitope mutation.3.Method of the cell cultivation and expansion is able to manufacture cell infusion product of HIV-specific CD8+T cell for clinical use.4.The Adoptive Cellular Therapy with HLA-A02 restricted HIV-specific CD8+T Cell studied caused only a few controllable adverse effects and no severe adverse effect,can be considered safe.5.The therapy could downregulate PD-1 and CD95/Fas expression on the surface of CD8+T cell,help the reconstitution of immunity.