HIV-1中国流行株的生物学表型与V3环序列变异相关性初步研究  

Biological characteristics of HIV-1 isolates circulating in China are linked to its env V3 loop sequence variability

作　　者：黑发欣[1] 洪坤学[1] 宋艳辉[1] 唐海丽[1] 彭虹[1] 徐建青[1] 邢辉[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病中心病毒免疫室,北京100050

出　　处：《中华医学杂志》2004年第23期1968-1972,共5页National Medical Journal of China

基　　金：国家重点基础研究发展规划基金资助项目 (G19990 5 410 7);美国NIH中国CIPRA基金资助项目(1U19AI5 1915 0 2 )

摘　　要：目的　分析我国HIV 1流行株的生物学表型及其与V3环序列变异的相关性。方法　采用传统的共培养方法从HIV 1感染者新鲜外周血单个核细胞 (PBMC)中分离病毒并在MT 2细胞上测定分离株的融合诱导性 ,用表达CD4和趋化因子受体CCR5或CXCR4的GHOST( 3)测定毒株的辅助受体利用情况 ,用巢式 聚合酶链反应 (nest PCR)扩增V3环及两侧区序列并对其序列进行测定 ,用GCG软件分析氨基酸序列。结果　在所分析的 5个原代毒株中 ,LTG0 2 13和LTG0 2 14为合胞体诱导型 (SI) ,利用CXCR4辅助受体 ;XJN0 0 2 1、XJN0 0 91和SHXDC0 0 4 1为非合胞体诱导型 (NSI) ,利用CCR5辅助受体。X4 /SI病毒和R5 /NSI病毒在V3环氨基酸序列方面 ,存在明显的区别。CCR5表型中国株第 8、11、18及第 2 5位氨基酸的共同基序为 :8 TXXS/GXXXXXXR/QXXXXXXE/D 2 5 ,CXCR4表型株在这些位置出现碱性氨基酸取代 (第 2 5位除外 ) ,引进正电荷。结论　HIV 1中国流行株的生物学表型与V3环序列变异密切相关 ,V3环上第 8、11、18及第 2 5位积累碱性氨基酸 (或由酸性氨基酸转变为中性氨基酸 ) ,可以使病毒由NSI/R5表型转变为SI/X4表型 。Objective To investigate the biological characteristics of the HIV-1 isolates circulating in China and to define the association of these properties with env V3 loop sequence variability. Methods Primary viruses were isolated from fresh peripheral blood mononuclear cells (PBMCs) using the traditional co-culture method and their capacity of inducing syncytium was tested in MT-2 cells; meanwhile, their coreceptor usage was determined with GHOST-cell lines which stably express CD4 and the chemokine receptor CCR5 or CXCR4. Furthermore, HIV-1 V3 and its flanking region sequences were amplified by nest-polymerase chain reaction (nest-PCR) and sequenced. A GCG software was used to translate the DNA sequences into polypeptide sequences.Results Five primary viral strains were isolated from 3 different regions in China. The isolates LTG0213 and LTG0214 induced syncytia in MT-2 cells and used CXCR4 as coreceptor. The isolates XJN0021, XJN0091, and SHXDC0041 did not induce syncytia and used CCR5 as coreceptor. There were obvious differences between X4/SI and R5/NSI viruses in env V3 loop sequences. A consensus motif at the positions 8, 11, 18, and 25 in V3 loop was identified as follows: a sequence as “8-TXXS/GXXXXXXR/QXXXXXXE/D-25” will predict the usage of CCR5 coreceptor; a sequence replacing these positions with basic amino acids (except position 25) will very likely predict the usage of CXCR4 coreceptor.Conclusion The biological characteristics of HIV isolates are linked to env V3 loop sequence variability: introducing basic amino acids (or translating from acidic amino acids into neutral amino acids) at the positions 8, 11, 18, and 25 in V3 loop will change viral strain′s biological phenotype from NSI/CCR5 to SI/CXCR4. The biological phenotype of HIV-1 can be predicted with V3 loop sequence analysis.

关 键 词：HIV-1 中国流行株 生物学表型 V3环序列变异 相关性 

分 类 号：R373[医药卫生—病原生物学]