Objective To assess the impact of the virus on the complementary determining region 3 (CDR3) length diversity of T cell receptor(TCR) Vβ repertoires of CD4+ T lymphocytes and to explore its association with viral load in individuals with HIV-1 infection. Methods The TCR repertoire was examined using spectratyping of CDR3 length diversity within CD4+ T cells in HIV infected and healthy adults. Separation of CD4+ T cells from peripheral blood mononuclear cells ( PBMCs) was carried out by using immunomagnetic beads coated with anti-CD4 antibody. Total RNAs from the purified CD4 + T lymphocytes were isolated and used to perform nested-PCR amplifications in CDR3 of 22 TCR gene families. CDR3 diversity and its association with viral load in individuals with HIV-1 infection were analyzed. Results An average diversity for all CDR3 profiles in CD4+ T cells from 25 HIV-infected individuals was significantly different as compared to 10 age-matched healthy donors (P＜0.05) with the HIV-infected individuals losing diversity in the CDR3 profiles. There was positive correlation between changes in TCR CDR3 diversity and viral load (r = 0. 494, P ＜ 0. 05). The changes in CDR3 length diversity of Vβ families in HIV-infected individuals, particular in Vβ8, Vβ22, Vβ23 were statistically different from the healthy controls. Conclusion HIV-1 infection might induce the loss of TCR Vp repertoire diversity and disrupt the CDR3 Gaussian distributions within CD4 + T cells. There should be positive correlation between changes in TCR CDR3 diversity and the viral load in HIV-1 infected patients.

Objective To explore the polymorphism of HLA class I alleles of HIV-infected former plasma donors and to investigate its impact on HIV-1 viral load in central China.Methods 106 subjects chronically infected with HIV-1 were recruited and HLA class I alleles were genotyped with PCR-SSP assay.HLA class I genotypes and haplotypes were determined and their association with plasma viral loads were analyzed.Gag-specific CTL responses were detected by an IFN-λ EUSPOT assay by using overlapping peptides,and their association with plasma viral loads were also analyzed.Results Subjects homozygous at HLA class I locus had higher plasma viral loads(P=0.0098);HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads(P=0.0004,0.0103,0.0058,0.0371 and 0.0006);an inverse correlation between p2p7p1p6-specific CTL responses and viral loads in subjects with HLA-A*30/B*13/Cw*06 haplotype as well as an inverse correlation between p17-specific CTL responses and viral loads in subjects with HLA-Cw*14 allele were observed.Conclusion HLA-A*30,-B*13,-Cw*06,-Cw*14 alleles and HLA-A*30/B*13/Cw*06 haplotype were associated with lower plasma viral loads and Gag-specific CTL responses restricted by these HLA alleles may contribute to the protection.

Objective To study biological characteristics of R5 tropic human immunodeficiency virus (HIV)-1 strains in different disease stage. Methods Primary clinical viruses were isolated from fresh peripheral blood mononuclear cells (PBMC) using co-culture methods; meanwhile, viral co receptor usage and infectivity were tested using flow cytometry on GHOST (3) cell lines,which expressed CD4 receptor and CC ehemokine receptor 5 (CCR5) or CXCR4 eoreceptor; to identified CCR5 tropic viruses(R5 tropic strains). Viral replication kinetics was detected in PBMCs. Plasma viral load was measured using an HIV-1 nucleotide fluorescence quantification assay kit. Results There were 22 individuals with HIV-1 subtype B' infection, in which 11 were CD4>0. 2 × 109/L and 11 were CD4≤0. 2 × 109/L. All isolated viruses used CCR5 coreceptor and therefore were HIV-1 R5 tropic strains. The infectivity of R5 tropic strains isolated from patients with CD4≤0.2 × 109/L was (7.392 7 ± 4. 584 2) % ; while the infectivity of R5 tropic strain from patients with CD4>0. 2 × 109/L was (2. 613 6 ± 1. 610 5)%. There were significant statistical difference(t= 3. 262, P<0.05). The possibility of viral replication became strong after the day 7 post-infection. There was a significant difference of viral replication between two groups in the day 7,10, 15 post-infection(t value was 3. 771, 2. 509 and 2. 260 respectively, P<0. 05). The possibility of viral replication was higher in CD4≤0.2 ×109/L group than that of CD4>0.2 × 109/L group. The logarithm of viral load was (5. 606 8 ± 0. 815 1 ) copies/mL in CD4≤0.2 × 109/L group and (4. 729 8 ± 0. 431 6) copies/mL in CD4> 0.2 × 109/L group. There was a significant difference between two groups(t = 3. 771 ; P<0.05). Conclusion Viral infection and replication are enhanced during progression of disease, even if viral coreceptor usage do not switch from CCR5 to CXCR4.

Objective To explore the association among viral load,CD4 count and neutralizing ac-tivity of plasma from chronic HIV-1 infected former blood donors in Anhui province, China. Methods 294 chronically HIV-1 B' infected individuals from former blood donors in Anhni province were enrolled, whose plasma and peripheral blood mononuelnar cells (PBMC) were isolated. The viral load and CD4 were detec-ted, and the neutralization activity of plasma against primary virus (1597) and lab-adapted strain (SF33) was detected by Luciferase Assay System based on TZM cell line, and 50% neutralizing level was calculated by the Reed-Mueneh method. Results Neutralizing activity responding against SF33 strain was higher than 1597 (P<0.05). There was a positive correlation between neutralizing concentration of plasma against SF33 and viral load (1g) (r = 0.191, P = 0.001), but there was no correlation between neutralizing concen-tration of plasma against 1597 and viral load(lg) (r = 0.097, P = 0.096). Conchtsion In chronic HIV-1 infectious people, neutralizing level against SF33 increases with viral replication, however, there is no asso-ciation of plasma neutralizing against 1597 replication with viral load.

Gene therapy is one of several approaches that are being tested in the search for an effective anti-HIV treatment. In this strategy, a "resistant" gene would be introduced into target cells, rendering them resistance to the infection of HIV. The HIV-1 Tat protein transactivate HIV-1 gene expression at the transcriptional level by interacting with its response element(TAR) in the long terminal repeat(LTR). Previously, we have shown that antisense polyTAR-Core RNAs can inhibit the transactivation of HIV-1 Tat protein in transiently transfected Jurkat cells. To determine whether this antisense polyTAR-Core RNAs could inhibit HIV-1 replication in CD4+ T cells, we transfected the antisense polyTAR-Core gene to MT4 cells and challenged them with HIV-1 SF33 strain. Levels of HIV-1 p24gag antigen were reduced more than 4-fold in cultures of the transduced MT4/LR cells infected with HIV-1SF33 strain. In contrast, cultures of nontransduced MT4 cells and control LX vector transduced MT4/LX cells infected with the same viruses had high levels of HIV-1 p24gag. Our work showed that antisense polyTAR-Core RNAs were able to inhibit HIV-1 replication in CD4+ T cells, and could be used as resistance gene in further studying for gene therapy against HIV-1.