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目的:阐明HIV-1感染者外周血中具有CD4+CD25nt/hiCD127lo特征的调节性T细胞(Treg)表面PD-1的表达水平与疾病进展的关系。方法:选取108名未经治疗的不同进展期的HIV-1感染者和27名健康人对照,采集静脉血,用Ficoll-Hypaque密度梯度离心法分离获得PBMC,加入PerCP-CD4抗体、FITC-CD25抗体、PE-CD127抗体和APC-PD-1抗体,经细胞表面四色染色、流式细胞术(FCM)分析Treg表面PD-1的表达;另将50 L全血加入Trucount绝对计数管,采用Multitest CD3/CD8/CD45/CD4试剂盒检测CD4+T细胞绝对数;分离静脉血血浆,NucliSens EasyQ测定血浆HIV-1病毒载量;实验数据采用SPSS14.0统计学软件分析处理。结果:HIV-1感染者Treg表面PD-1表达水平显著高于健康人(5.33%±2.24%vs1.72%±0.65%,P<0.01);AIDS期(7.87%±2.23%)明显高于进展期(5.21%±1.72%,P<0.05)和新近感染者(3.22%±1.01%,P<0.05);HIV-1感染者Treg表面PD-1表达水平与血浆中的HIV-1病毒载量和CD4+T细胞绝对数密切相关。结论:首次证实HIV-1感染者外周血中Treg表面PD-1表达增加,且表达水平与病程进展相关。该结果为进一步揭示HIV-1感染中Treg的效应机制、探索新的免疫治疗方案提供了理论及实验依据。
Objective: To elucidate the relationship between the expression of PD-1 on the surface of regulatory T cells (Treg) with CD4 + CD25nt / hiCD127lo in peripheral blood of HIV-1 infected patients and the progression of the disease. Methods: 108 untreated patients with HIV-1 and 27 healthy controls were enrolled in this study. Venous blood was collected and PBMCs were isolated by Ficoll-Hypaque density gradient centrifugation. PerCP-CD4 antibody and FITC-CD25 PE-CD127 antibody and APC-PD-1 antibody. The expression of PD-1 on Treg surface was analyzed by flow cytometry (FCM). Another 50 L whole blood was added into Trucount absolute counting tube. CD4 + T cells were detected by Multitest CD3 / CD8 / CD45 / CD4 kit; venous plasma was separated and NucliSens EasyQ was used to detect the plasma HIV-1 viral load. The experimental data were analyzed by SPSS14.0 statistical software. Results: The expression of PD-1 on Tregs in HIV-1 infected patients was significantly higher than that in healthy controls (5.33% ± 2.24% vs 1.72% ± 0.65%, P <0.01), and was significantly higher in AIDS (7.87% ± 2.23% (5.21% ± 1.72%, P <0.05) and newly infected patients (3.22% ± 1.01%, P <0.05). The expression of PD-1 on Treg surface in HIV-1 infected patients was significantly higher than that in HIV- The amount of CD4 + T cells is closely related to the absolute number. Conclusions: It was confirmed for the first time that the expression of PD-1 on Treg surface in peripheral blood of HIV-1 infected patients was increased, and the expression of PD-1 was correlated with the course of the disease. The results provide a theoretical and experimental basis for further revealing the mechanism of Treg in HIV-1 infection and exploring new immunotherapy programs.