HIV-1B′亚型毒株抵抗VRC01抗体中和作用的机制研究  

Molecular mechanism of resistance to VRC01 neutralization in HIV-1 subtype B′strains

作　　者：张岱[1,2] 刘真[3] 王炜[1] 侯佳利[2] 郝彦玲[2] 任伟宏[1] 任莉[2] Zhang Dai;Liu Zhen;Wang Wei;Hou Jiali;Hao Yanling;Ren Weihong;Ren Li(Department of Clinical Laboratory,the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;AIDS Research Center,the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院检验科,郑州450000 [2]中国疾病预防控制中心性病艾滋病预防控制中心,北京102206 [3]河南中医药大学第一附属医院艾滋病研究中心,郑州450000

出　　处：《中华微生物学和免疫学杂志》2021年第9期692-697,共6页Chinese Journal of Microbiology and Immunology

基　　金：政府间国际科技创新合作重点专项(2016YFE0107600);河南省中医药科学研究专项课题(2018JDZX065,2019JDZX2090);河南省科技攻关计划(202102310178)。

摘　　要：目的探讨具有广谱中和活性的患者DRVI01血浆来源的HIV-1B′亚型毒株抵抗VRC01抗体中和的机制。方法比较同期感染相同亚型且对VRC01抗体敏感的病毒序列,结合文献报道筛选可能影响VRC01中和作用的关键氨基酸,通过定点突变、不同来源膜蛋白序列交换,验证这些位点氨基酸对VRC01抗体中和作用的影响。结果位于gp120的LoopD区E279D、R282K和V5区N460A、N463Q单点突变,逆转了病毒对VRC01中和敏感性。上述2个或3个位点联合突变后的毒株与单点突变毒株比较,对VRC01抗体中和敏感性明显增强。与文献报道不同,N276糖基化位点突变并未改变毒株对VRC01的敏感性。结论具有广谱中和活性患者DRV01血浆来源的HIV-1B′亚型毒株主要通过LoopD区D279、K282和V5区N460、N463突变,抵抗VRC01抗体的中和作用。Objective To investigate the molecular mechanism of VRC01 resistance in HIV-1 subtype B′strains isolated from a patient(DRVI01)with broadly neutralizing antibody(bNAb).Methods Sequences of the HIV-1 subtype B′strains isolated from patient DRVI01 were compared with those of HIV-1 subtype B′strains that were isolated at the same time but sensitive to VRC01 antibody.Key amino acids that might affect the neutralization of VRC01 were selected according to literature reports.Effects of the selected amino acids on VRC01 neutralization were verified by site-directed mutation and sequence exchange of membrane proteins from different patients.Results Single-point mutations of E279D and R282K in LoopD region and N460A and N463Q in V5 region reversed the viral sensitivity to VRC01 neutralization.Combined mutations in two or three above-mentioned sites significantly increased the viral sensitivity to VRC01 antibody compared with single-point mutations.Contrary to literature reports,the glycosylation site mutation of N276 had no influence on the viral sensitivity to VRC01.Conclusions HIV-1 subtype B′strains isolated from patient DRV01 with bNAb carried the mutations of D279 and K282 in LoopD region and N460 and N463 in V5 region,resulting in resistance to VRC01 antibody.

关 键 词：HIV-1 膜蛋白 中和抗体 定点突变 

分 类 号：R392[医药卫生—免疫学]