R5嗜性的人类免疫缺陷病毒-1在疾病不同阶段的生物学特性  

The biologic characteristics of human immunodeficiency virus-1 subtype B＇ R5 tropic strains in different disease stage

作　　者：郭艳芳[1] 马丽英[1] 张跃新[2] 袁林[1] 孙坚萍[1] 徐维四[1] 赵全壁[1] 屈水令[1] 黄洋[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室,北京100050 [2]新疆医科大学第一附属医院感染科

出　　处：《中华传染病杂志》2008年第7期425-429,共5页Chinese Journal of Infectious Diseases

基　　金：国家973计划资助项目（2005CB523103）;中国综合性艾滋病研究资助项目（U19AIS1915-03）;国家863计划资助项目（2006AA03Z323）

摘　　要：目的研究HIV-1感染者不同时期分离的R5毒株的生物学特性。方法采用传统的共培养方法分离并培养HIV-1，用表达CD4和CC趋化因子受体5（CCR5）或CXC趋化因子受体4（CXCR4）的GHOST细胞系，通过流式细胞仪测定病毒辅助受体的利用和感染性，从而判断所分离毒株的CCR5嗜型（R5型毒株）；使用2ng P24病毒量感染正常人分离的外周血单个核细胞（PBMC）．ELISA法检测第1、3、5、7、10、15天的HIV-1 P24抗原．反映病毒复制能力；采用HIV-1核酸荧光定量检测试剂盒测定血浆病毒载量。数据分析采用t检验。结果HIV-1B’亚型感染者22例，其中CD4^＋细胞〉0．2×10^9／I。和CD4^＋细胞≤0．2×10^9／L各11例；昕分离的病毒仅利用CCR5辅助受体，均为R5型毒株．感染性的结果显示．来自CD4^＋细胞≤0．2×10^9／L的11株R5毒株的感染性为（7．3927±d．5842）％，而CD4^＋细胞〉0．2×10^9／L的为（2．6136±1．6105）％，差异有统计学意义（t=3．262，P〈0．05）；两组病毒复制滴度在第7天开始明显上升，培养第7、10、15天，两组病毒复制动力学差异有统计学意义（t值分别为3．771、2．509和2．260；P〈0．05），CD4^＋细胞≤0．2×10^9／L的R5毒株的复制能力较CD4^＋细胞〉0．2×10^9／L的明显增强；CD4^＋细胞≤0．2×10^9／L R5型毒株的病毒载量的对数值为（5．6068±0．8151）拷贝／mL，CD4^＋细胞〉0．2×10^9／L的为（4．7298±0．4316）拷贝／mL，两组差异有统计学意义（t=3．771，P〈0．05）。结论疾病进展过程中。即使病毒的辅助受体利用未从CCR5转变为CXCR4，但病毒的感染性和复制能力已有明显改变。Objective To study biological characteristics of R5 tropic human immunodeficiency virus （HIV）-1 strains in different disease stage. Methods Primary clinical viruses were isolated from fresh peripheral blood mononuclear cells （PBMC） using co-culture methods; meanwhile, viral co receptor usage and infectivity were tested using flow cytometry on GHOST （3） cell lines, which expressed CD4 receptor and CC chemokine receptor 5 （CCRS） or CXCR4 coreceptor; to identified CCR5 tropic viruses（R5 tropic strains）, Viral replication kinetics was detected in PBMCs. Plasma viral load was measured using an HIV-1 nucleotidc fluorescence quantifieation assay kit. Results There were 22 individuals with HIV-1 subtype B＇ infection, in which 11 were CD4 〉 0.2 × 10^9/L and 11 were CD4 ≤ 0. 2 × 10^9 / L. All isolated viruses used CCR5 coreceptor and therefore were HIV-1 R5 tropic strains. The infectivity of R5 tropic strains isolated from patients with CD4≤0.2 × 10^9/L was （7. strain from patients with CD4〉0. 2 × 10^9/1. was 392 7±4. 584 2） % ; while the infectivity of R5 tropic （2. 613 6±1. 610 5）%. There were significant statistical difference（t=3. 262, P〈0.05）. The possibility of viral replication became strong after the day 7 post-infection. There was a significant difference of viral replication between two groups in the day 7,10,15 post-infection（t value was 3. 771, 2. 509 and 2. 260 respectively, P〈 0. 05）. The possibility of viral replication was higher in CD4≤0.2 × 10^9/L group than that of CD4〉0.2 × 10^9/L group. The logarithm of viral load was （5. 606 8±0. 815 1） copies/mL in CD4≤0.2 × 10^9/L group and （4. 729 8±0. 431 6） copies/mL in CD4 〉 0. 2 × 10^9/L group. There was a significant difference between two groups（t= 3. 771 ; P〈 0.05）. Conclusion Viral infection and replicatioh are enhanced during progression of disease, even if viral coreceptor usage do not switch from CCR5 to CXCR4.

关 键 词：HIV-1 受体 细胞表面 受体 CCR5 受体 CXCR4 

分 类 号：R51[医药卫生—内科学] R3[医药卫生—临床医学]