早期HIV-1感染者单核细胞对TLR配体反应性研究
梁华1，沈弢2
基金项目：教育部博士点基金(20100001120047)；国家自然科学基金项目(81271826, 31100126) 作者简介：梁华，女，医学博士，主要研究方向为病毒免疫学 通信联系人：沈弢(1971-)男，副教授，主要研究方向为HCV的病毒学和免疫学. 

（1. 中国疾病预防控制中心性病艾滋病预防控制中心病毒免疫室； 5 2. 北京大学医学部基础医学院病原生物学系）
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，采血分离PBMC后纯化出单核细胞，流式细胞术检测单核细胞表面活10 化/抑制分子表达。用LPS和Pam3CSK4分别刺激单核细胞，培养20h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15 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。
关键词：人类免疫缺陷病毒；单核细胞；TLR2；TLR4 20
中图分类号：[R394.5]
Responses of Monocytes to TLR Ligands in Primary HIV-1 Infection
Hua Liang1, Tao Shen2 25
(1. National center for HIV/AIDS control and prevention, China CDC; 2. Department of Microbiology, Peking University Health Science Center)
Abstract: Objective To explore the maturation status of monocytes and the responses of monocytes to TLR ligands in primary HIV-1 infection, and to understand the association between functional status of monocytes and disease progression in primary HIV-1 infection. Methods 30 Monocytes were isolated from PBMC of 35 primary HIV-1 infected subjects. Activate/inhibitory markers expression on monocytes were analyzed by flow cytometry. Monocytes were stimulated with LPS and Pam3CSK4 and cultured for 20h. Activate/inhibitory markers expression on monocytes were analyzed by flow cytometry and secretions of proinflammatory cytokines by stimulated monocytes were detected by ELISA. Results In primary HIV-1 infection, activate 35 markers CD80, CD86, CD40, and inhibitory marker PD-L1 expression on monocytes were increased (P<0.001 except for CD86 P=0.01). CD40 expression was positively correlated with plasma viral load (P<0.001，R=0.553). After stimulated with LPS and Pam3CSK4, the increases of HLA-DR, CD80, CD86, and PD-L1 expression were lower in primary HIV-1 infection compared with healthy controls (P<0.001)，and secretions of proinflammatory cytokines TNF-α40 （LPS：P=0.004，Pam3CSK4：P=0.012）and IL-6（LPS：P=0.006）were enhanced in monocytes of primary HIV-1 infection. Conclusion In primary HIV-1 infection, monocytes were activated and secreted higher levels of proinflammatory cytokines after TLR ligands stimulation, suggesting a possible role of monocytes in microbial translocation and immune activation in HIV-1 infection.
Key words: human immunodeficiency virus; monocytes; Toll like receptor 2; Toll like receptor 4