早期 HIV-1感染者单核细胞对 TLR 配体反应性研究

摘要

目的：探讨HIV-1早期感染者体内单核细胞的成熟状态和对TLR配体的反应特点，了解HIV-1感染早期单核细胞功能状态和与疾病进展状态的相关性。方法选取35例HIV-1早期感染者和13例HIV阴性的健康人，采血分离外周血单个核细胞（ PBMC ）后纯化出单核细胞，流式细胞术检测单核细胞表面活化／抑制分子表达。用脂多糖（ LPS）和Pam3CSK4分别刺激单核细胞，培养20 h后，流式细胞术检测单核细胞表面活化／抑制分子表达，ELISA方法检测培养上清中促炎细胞因子IL-1β、TNF-α、IL-6的分泌水平。结果 HIV-1早期感染者，单核细胞表面活化分子CD80、CD86、CD40和抑制分子PD-L1表达均升高（除CD86 P＝0．01外，P均＜0．001），CD40表达水平与血浆病毒载量正相关（P＜0．001，r＝0．553）。单核细胞受LPS和Pam3CSK4刺激后，活化成熟能力降低，HLA-DR、CD80、CD86、PD-L1表达增加均较健康对照低（P＜0．001），而促炎细胞因子 TNF-α（LPS：P＝0．004，Pam3CSK4：P＝0．012）和IL-6分泌（LPS：P＝0．006）增加。结论在HIV-1感染早期，单核细胞即处于免疫活化状态，受TLR配体刺激后促炎细胞因子分泌增加可能与HIV-1感染后的微生物移位和免疫活化相关。%Objective To investigate the maturation status of monocytes and their responses to the stimulation of toll like receptor (TLR) ligands during primary HIV-1 infection, and to further understand the correlation between functional status of monocytes and disease progression during primary HIV -1 infection. Methods Peripheral blood mononuclear cells ( PBMCs) were collected from 35 subjects with primary HIV-1 infection and 13 HIV-negative healthy subjects to isolate monocytes .Monocytes were stimulated with LPS and Pam3CSK4, respectively, and cultured for 20 hours.The expression of activaion/inhibitory markers on monocytes were analyzed by flow cytometry before and after stimulation .The secretion of proinflammatory cy-tokines ( IL-1β, TNF-αand IL-6) by stimulated monocytes were detected by ELISA .Results The expres-sion of activation markers CD80, CD86, CD40 and inhibitory marker PD-L1 on monocytes were increased in subjects with primary HIV-1 infection (P<0.001 except for CD86 P=0.01).The level of CD40 was posi-tively correlated with viral load in plasma (P<0.001, R=0.553).Compared with control group, primary HIV-1 infection group showed a less increase in the expression of HLA-DR, CD80, CD86 and PD-L1 on monocytes after stimulation with LPS and Pam3CSK4 (P<0.001), but the secretion of proinflammatory cyto-kines TNF-α(LPS:P=0.004, Pam3CSK4:P=0.012) and IL-6 (LPS:P=0.006) were enhanced in mono-cytes from patients with primary HIV-1 infection.Conclusion Monocytes were activated during primary HIV-1 infection.They secreted higher level of proinflammatory cytokines after stimulation with TLR ligands , indicating monocytes might play a role in microbial translocation and immune activation during HIV -1 infection .