Abstract：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. Methods The 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. Results Totally 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. Conclusions The 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.
李贺, 杜波, 田晓东, 刘文新, 姚文清. 阜新市HIV-1env基因C2~V4区序列特征分析[J]. 预防医学, 2020, 32(10): 983-986.
LI He, DU Bo, TIAN Xiaodong, LIU Wenxin, YAO Wenqing. Sequence analysis of C2-V4 of HIV-1 env genes in Fuxin. Preventive Medicine, 2020, 32(10): 983-986.
 董万强,姚能,徐庆刚.亚洲主要流行HIV-1 Gp120及其5个高变区多态性分析[J].生物学杂志,2013,30(1):9-13.  SCHEID J F,MOUQUET H,FELDHAHN N,et al.Broad diversity of neutralizing antibodies isolated from memory B cells in HIV-infected individuals[J].Nature,2009,458(7238):636-640.  兰芸,胡凤玉,李凌华,等.广州市男男性行为HIV-1感染者主要流行株Gp120序列变异分析[J].中华实验和临床病毒学杂志,2019,33(3):291-296.  HEDSKOG C,BRODIN J,HEDDINI A,et al.Longitudinal ultradeep characterization of HIV type 1 R5 and X4 subpopulations in patients followed from primary infection to coreceptor switch[J].AIDS Res Hum Retroviruses,2013,29(9):1237-1244.  叶景荣,邢辉,刘海林,等.北京市2006年HIV-1流行毒株的gag和env基因序列测定及亚型分析[J].中华流行病学杂志,2007,28(6):586-588.  LENGAUER T,SANDER O,SIERRA S,et al.Bioinformatics prediction of HIV coreceptor usage[J].Nat Biotechnol,2007,25(12):1407-1410.  常晓云,李海宁,王馨仪,等.黑龙江省HIV-1型流行株CRE01_AE env基因序列特征和传播簇分析[J].国际免疫学杂志,2018,41(2):142-147.  王敏连,梁冰玉,叶力,等.广西HIV-1流行株env基因C2V3区序列特征和亚型研究[J].中华疾病控制杂志,2015,19(12):1191-1194.  何瑞英,廖宝林,袁小珍,等.广东省人类免疫缺陷病毒1型CRF01_AE毒株膜区V3环氨基酸序列多态性分析[J].中国热带医学,2018,18(10):984-990.  BOONCHAWALIT S,JULLAKSORN D,UTTIYOUNG J,et al.Molecular evolution of HIV-1 CRF01_AE Env in Thai patients[J].PLoS One,2011,6(11):e27098.  ZHANG Y,LU L,BA L,et al.Dominance of HIV-1 subtype CRF01_AE in sexually acquired cases leads to a new epidemic in Yunnan province of China[J].PLoS Med,2006,3(11):e443.  劳飞翔,莫实德,邓培雪,等.广西防城港市HIV-1流行株env基因C2V3区序列特征及亚型分析[J].医学动物防制,2019,35(7):624-628.  JOSHI A,NYAKERIGA A M,RAVI R,et al.HIV ENV glycoprotein-mediated bystander apoptosis depends on expression of the CCR5 co-receptor at the cell surface and ENV fusogenic activity[J].J Biol Chem,2011,286(42):36404-36413.  张燕,赵广录,石向东,等.深圳市HIV-1主要流行株外膜蛋白V3环氨基酸变异分析[J].中国热带医学,2019,19(7):638-641.  PIYASIRISILP S,MCCUTCHAN F E,CARR J K,et al.A recent outbreak of human immunodeficiency virus type 1 infection in Southern China was initiated by two highly homogeneous,geographically separated strains,circulating recombinant form AE and a novel BC recombinant[J].J Virol,2000,74(23):11286-11295.  MILICH L,MARGOLIN B H,SWANSTROM R,et al.Patterns of amino acid variability in NSI-like and SI-like V3 sequences and a linked change in the CD4-binding domain of the HIV-1 Env protein[J].Virology,1997,239(1):108-118.  ZHOU T Q,XU L,DEY B,et al.Structural definition of a conserved neutralization epitope on HIV-1 gp120[J].Nature,2007,445(7129):732-737.  WU X,ZHOU T,O'DELL S,et al.Mechanism of human immunodeficiency virus type 1 resistance to monoclonal antibody B12 that effectively targets the site of CD4 attachment[J].J Virol,2009,83(21):10892-10907.  KWONG P D,DOYLE M L,CASPER D J,et al.HIV-1 evades antibody-mediated neutralization through conformational masking of receptor-binding sites[J].Nature,2002,420(6916):678-682.  SCANLAN C N,PANTOPHLET R,WORMALD M R,et al.The broadly neutralizing anti-human immunodeficiency virus type 1 antibody 2G12 recognizes a cluster of alpha 1→2 mannose residues on the outer face of gp120[J].J Virol,2002,76(14):7306-7321.