DC-SIGN诱导的信号通路在HIV-1前病毒活化中的作用  

Role of DC-SIGN induced signaling pathway in the activation of HIV-1 virus

作　　者：李杰[1] 靳昌忠[2] 程林芳[2] 刘福民[2] 吴南屏[2] LI jie JIN Chang-zhong CHENG Lin-fang LIU Fu-min WU Nan-ping(Department of Infectious Disease, The Second Affiliated Hospitlal （ Yuying Children, s Hospital） of Wenzhou Medical University, Wenzhou 325027 China State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, College of Medicine of Zhejiang University, Hangzhou 310003, China)

机构地区：[1]温州医科大学附属第二医院感染内科,浙江温州325027 [2]浙江大学医学院附属第一医院传染病诊治国家重点实验室,浙江杭州310003

出　　处：《中国生化药物杂志》2016年第12期41-45,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81402726);浙江省公益性技术应用研究计划(2015C33183)

摘　　要：目的研究HIV-1 gp120蛋白与DC-SIGN结合对HIV-1前病毒的活化作用及其信号通路机制。方法将DC-SIGN表达质粒和HIV-1 5’末端重复序列(5’LTR)报告质粒转染293T细胞,以HIV-1 gp120蛋白、野生型HIV-1、VSV-G-p NL4.3假病毒分别刺激,检测HIV-1 5’LTR的活性。慢性感染HIV-1的CEM-Bru细胞转染DC-SIGN表达质粒,以HIV-1 gp120蛋白刺激,检测HIV-1Tat mRNA和HIV-1 p24蛋白表达水平。特异性抑制ERK、P38和NF-κB信号通路,再用gp120刺激DC-SIGN(+)CEM-Bru细胞,检测HIV-1前病毒的活化。结果在DC-SIGN(+)293T细胞中,HIV-1 5'LTR可以被HIV-1 gp120激活。HIV-1 gp120蛋白及DC-SIGN刺激DC-SIGN(+)CEM-Bru后,HIV-1 Tat mRNA和HIV-1 p24蛋白表达水平明显升高,提示其早期和晚期HIV-1前病毒复制,用抗体阻断DC-SIGN可抑制潜伏HIV-1活化。HIV-1 gp120/DC-SIGN通过NF-κB信号通路在激活潜伏HIV-1前病毒。结论 HIV-1gp120可通过结合DC-SIGN激活NF-κB信号通路介导HIV-1前病毒活化。Objective To explore the mechanism of latent human immunodeficiency ciency virus type 1（ HIV-1） infection is unclear,especially in dendritic cells（ DC）. We hypothesized that DC-specific intercellular adhesion molecule-3-grabbing non-integrin（ DC-SIGN） binds with HIV-1 may activate HIV-1 provirus. Methods We generated a model by transfecting 293 T cells with a DC-SIGN expression plasmid and a HIV-1 5＇ long terminal repeat（ LTR） reporter plasmid,and then stimulated the 293 T cells with HIV-1 gp120 protein,wild-type HIV-1 and VSV-G-p NL4. 3 pseudotype virus（ without gp120 protein）. CEM-Bru cells were transfected with the DC-SIGN expression plasmid and stimulated by HIV-1 gp120 protein. Then HIV-1replication was detected. The involvement of the ERK,p38 and NF-κB pathways signaling in this response were determined by inhibiting the pathways specifically and detecting the phosphorylation of the signaling kinase. Results The HIV-1 5＇ LTR was reactivated by HIV-1 gp120 in DC-SIGNexpressing 293 T cells. After HIV-1 gp120 protein stimulation of the mold of CEM-Bru cells,the increasing expression of HIV-1 Tat mRNA and HIV-1p24,which implies early and late HIV-1 provirus replication was reactivated by the HIV-1 gp120 / DC-SIGN stimulation. HIV-1 gp120 / DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway. Conclusion HIV-1 gp120 / DC-SIGN stimulation reactivates latent HIV-1provirus via the NF-κB signal pathway.

关 键 词：HIV-1 DC-SIGN 病毒潜伏 信号通路 NF-ΚB 

分 类 号：R322.2[医药卫生—人体解剖和组织胚胎学] R512.91[医药卫生—基础医学]