高效抗逆转录病毒治疗对HIV感染者外周血病毒储存库的影响

Effect of highly active antiretroviral therapy on viral reservoir in peripheral blood lymphocytes of HIV infections

目的 探讨接受与未接受HAART的HIV-1感染者外周血淋巴细胞亚群内HIV-1病毒储存库的差异.方法 采集及分离21例未经HAART和22例经HAART的HIV-1感染者外周血单核细胞(PBMC),采用磁珠分选法分选出CD4+T细胞、CD8+T细胞和非T细胞亚群,采用高敏感度实时荧光定量PCR法检测患者外周血HIV-1 RNA及上述3类分选后淋巴细胞亚群内HIV-1整合DNA容量(Alu-PCR方法);流式细胞术检测患者血中的CD3+、CD4+、CD8+T细胞数.结果 两组非T细胞中HIV-1整合DNA容量比较差异有统计学意义(Z=-3.326,P＜0.01),CD4+和CD8+T细胞中的HIV-1整合DNA容量比较差异均无统计学意义(Z=-1.871、-1.360,P＞0.05).未HAART组患者CD4+T细胞中HIV-1整合DNA容量与外周血HIV-1 RNA水平呈正相关(r=0.594,P＜0.05),与CD4+T细胞数和CD4+/CD8+比值呈负相关(r=-0.547、-0.482,P＜0.05),与CD8+T细胞数无相关性(P＞0.05);CD8+T细胞和非T细胞中HIV-1整合DNA与CD4+T、CD8+T细胞数和CD4+/CD8+比值均无相关性(P均＞0.05).HAART组患者T细胞亚群中HIV-1整合DNA容量与CD4+T、CD8+T细胞数和CD4+/CD8+比值均无相关性(P均＞0.05).结论 HIV-1感染者病毒储存库主要存在于外周血CD4+T细胞中,少量存在于CD8+T细胞和非T细胞中;接受HAART可能并不能有效减少T细胞中的病毒储存库.

Objective To explore the difference of HIV-1 latent reservoir in peripheral blood lymphocyte subsets of HIV-1 infected patients receiving HAART and HAART-na(i)ve.Methods The peripheral blood mononuclear cells (PBMCs) of HIV-1 infected patients were collected and isolated from 21 cases with HAART-na(i)ve and 22 cases with HAART.The CD4+T lymphocytes,CD8+T lymphocytes and non-T lymphocyte subsets in PBMCs were sorted by magnetic bead cell sorting.HIV-1 RNA in peripheral blood and HIV-1 integrated DNA in above three lymphocyte subsets were detected by high-sensitive real-time fluorescence quantitative PCR and Alu-PCR,respectively.CD3+,CD4+,CD8+T lymphocyte numbers were counted by flow cytometry.Results The difference between HIV-1 integrated DNA of non-T lymphocytes in the two groups had statistical significance (Z=-3.326,P＜0.01),while the differences of HIV-1 integrated DNA in CD4+T lymphocytes and CD8+T lymphocytes had no statistical significances (Z=-1.871 and -1.360,P both ＞0.05).In HAART-na(i)ve group,HIV-1 integrated DNA in CD4+T lymphocytes had a positive correlation with HIV-1 RNA in peripheral blood (r=0.594,P＜0.05),negative correlations with CD4+T lymphocyte counts and CD4+/CD8+ ratio (r=-0.547 and-0.482,P both＜0.05) and no correlation with CD8+T lymphocyte counts(P＞0.05).There was no correlation among HIV-1 integrated DNA in CD8+T lymphocytes and non-T lymphocytes,CD4+T and CD8+T lymphocyte counts and CD4+/CD8+ ratio (P all ＞ 0.05).There was no correlation among HIV-1 integrated DNA in the T lymphocytes of HAART group,CD4+T and CD8+T lymphocyte counts and CD4+/CD8+ ratio(P all ＞ 0.05).Conclusions HIV-1 latent reservoir mainly exists in peripheral blood CD4+T lymphocytes,with a small amount in peripheral blood CD8+T lymphocytes and non-T lymphocytes.HAART may not reduce the amount of HIV-1 latentreservoir in T lymphocytes.