阜新市HIV-1 env基因C2-V4区序列特征分析  

Sequence analysis of C2-V4 of HIV-1 env genes in Fuxin

作　　者：李贺 杜波 田晓东 刘文新 姚文清[2] LI He;DU Bo;TIAN Xiaodong;LIU Wenxin;YAO Wenqing(Fuxin Health Service Center,Fuxin,Liaoning 123000,China)

机构地区：[1]阜新市卫生健康服务中心,辽宁阜新123000 [2]辽宁省疾病预防控制中心

出　　处：《预防医学》2020年第10期983-986,共4页CHINA PREVENTIVE MEDICINE JOURNAL

基　　金：国家科技重大专项艾滋病和病毒性肝炎等重大传染病防治项目(2017ZX10103007)。

摘　　要：目的分析阜新市艾滋病病毒1型(HIV-1) env基因C2-V4区序列及代表性广谱单克隆中和抗体的表位变异,为HIV-1变异趋势研究及阐明V3环与病毒生物学特征提供依据。方法采集2018—2019年在阜新市卫生健康服务中心确证阳性的112例HIV-1感染者全血标本,采用巢式PCR扩增env基因C2-V4区核苷酸序列并测序,采用MEGA软件鉴定分型,采用HIV Database等软件分析V3环顶端四肽、辅助受体、净电荷和特征性氨基酸。结果获得101条有效基因序列,发现5种V3环顶端四肽类型,其中GPGQ 77条,占76.24%,存在于CRF01_AE、CRF07_BC、CRF65_cpx和G亚型;GPGR 19条,占18.81%,存在于CRF01_AE、CRF07_BC和B亚型;GPGH 3条,GPGK和GPGA各1条,均只存在于CRF01_AE亚型。辅助受体主要使用CCR5,84条占83.17%。CRF01_AE、CRF07_BC、B、CRF65_cpx和G亚型V3环净电荷数分别为3.28±1.17、3.22±0.92、4.25±0.83、2.50±0.50和3。结合b12和VRC01中和抗体表位氨基酸位点突变率为0-9.90%;295位和332位氨基酸N-糖基化位点缺失率分别为18.81%和14.85%,未发现两者同时缺失的情况。结论 2018—2019年阜新市HIV-1毒株主要是以CCR5为辅助受体的巨噬细胞嗜性非合胞体诱导型病毒,V3环顶端四肽以GPGQ为主,复制速度较慢,逃逸中和抗体能力较低。Objective To learn the sequence characteristics of C2-V4 of HIV-1 env genes and the epitope variation of representative broadly monoclonal neutralizing antibody in Fuxin, so as to provide evidence for the HIV-1 variation trend and the biological characteristics of V3 loop.MethodsThe whole blood samples of 112 HIV-1 cases in Fuxin Health Service Center from 2018 to 2019 were collected and the DNA was extracted. The C2-V4 of env genes were amplified by nested-PCR and the PCR products were subjected to sequencing. The bioinformatics analysis was carried out using MEGA software, and the V3-tip motifs, co-receptors, net charge and characteristic amino acids were analyzed using HIV Database.ResultsTotally 101 effective gene sequences were obtained, and 5 types of V3-tip motifs were found. Among them, 77 pieces of GPGQ(76.24%) were found in CRF01_AE, CRF07_BC, CRF65_cpx and G subtypes;19 pieces of GPGR( 18.81%) were found in CRF01_AE, CRF07_BC and B subtypes;3 pieces of GPGH, 1 piece of GPGK and 1 piece of GPGA were only found in CRF01_ AE subtype. The co-receptor was mainly CCR5( 84,83.17%). The net charge numbers of V3 loops in CRF01_ AE, CRF07_ BC, B, CRF65_cpx and G were 3.28±1.17,3.22±0.92, 4.25±0.83, 2.50±0.50 and 3, respectively. The mutation rates of neutralizing antibodies binding b12 and VRC01 were 0-9.90%. The deletion rates of N-glycosylation sites of 295 and 332 were 18.81% and 14.85%, without the loss of both sites.ConclusionThe HIV-1 strains in Fuxin from 2018 to 2019 are macrophage-tropic and non-syncytium-inducing, with GPGQ as the main type of V3-tip motif, CCR5 as the main co-receptor, slow replication and low ability to escape neutralizing antibodies.

关 键 词：艾滋病病毒1型 ENV基因 变异 包膜蛋白 V3环 

分 类 号：R512.91[医药卫生—内科学]