深圳市HIV-1主要流行株外膜蛋白V3环氨基酸变异分析

Mutation of envelop protein V3 loop in HIV-1 predominant subtype in Shenzhen

目的了解深圳市HIV-1主要流行株膜蛋白V3环氨基酸变异情况。方法收集深圳市1996—2010年HIV确认阳性样本389份,应用巢式聚合酶链反应(Nested-PCR)技术,对样本膜蛋白(Env)基因V3-V4区进行扩增,并对核苷酸序列进行测定,判定亚型结果。对获得的CRF01_AE、CRF_BC、B’和B重组株/亚型的核酸序列进行比对和翻译,分析其V3环氨基酸变异情况。结果 227例CRF01_AE重组株V3环顶端四肽主要为GPGQ(184/227,81.1%),199例(87.7%)可预测使用CCR5辅助受体,12例(5.3%)使用CXCR4辅助受体;55例CRF_BC重组株V3环顶端四肽主要为GPGQ(54/55,98.2%),50例(90.9%)可预测使用CCR5辅助受体,5例(9.1%)不能预测;49例B’亚型毒株V3环顶端四肽主要为GPGR(39/49,79.6%),25例(51.0%)可预测使用CCR5辅助受体,3例(6.1%)使用CXCR4辅助受体,21例(42.9%)不能预测;10例B亚型毒株V3环顶端四肽主要为GPGR(5/10,50.0%),8例(80.0%)可预测使用CCR5辅助受体,2例(20.0%)使用CXCR4辅助受体。CRF01_AE、CRF_BC、B’和B亚型毒株V3环净电荷数分别为3.29±0.97(n=227)、3.11±0.42(n=55)、3.73±0.93(n=49)和3.50±0.97(n=10)。结论深圳市HIV-1 V3 环顶端四肽主要为GPGQ和GPGR,病毒辅助受体使用以及表型预测结果表明深圳市HIV-1的流行仍以复制速度较慢的非合胞体诱导型病毒为主。

Objective To investigate the characteristics of human immunodeficiency virus (HIV-1) V3 loop amino acid mutations in Shenzhen. Methods Totally 389 HIV-1 positive plasma were collected in Shenzhen from 1996 to 2010.V3-V4 fragments of HIV-l Env gene were amplified by nested-PCR from RNA. Subtype analyses were performed on the nucleotide sequence data. CRF01_AE, CRF_BC, B’ and B strains sequences were aligned and translated, and analyzed the V3 loop amino acid mutations. Results Totally 227 CRF01_AE strain samples showed the main type of V3 loop central motif was GPGQ(184/227,81.1%), 199 cases(87.7%) were predicted to be CCR5-using, 12 cases (5.3%) were predicted to be using CXCR4. 55 CRF_BC recombinant strain samples showed the main type of V3 loop central motif was GPGQ(54/55,98.2%), 50 cases(90.9%) were predicted to be CCR5-using, 5 cases (9.1%) could not be predicted. 49 B’ strain samples showed the main type of V3 loop central motif was GPGR(39/49,79.6%), 25 cases(51.0%) were predicted to be CCR5-using, 3 cases(6.1%) were predicted to be using CXCR4 and 21 cases (42.9%) could not be predicted. 10 B strain samples showed the main type of V3 loop central motif was GPGR(5/10,50.0%), 8 cases(80.0%) were predicted to be CCR5-using, 2 cases(20.0%) were predicted to be using CXCR4. The electrostatic charge of CRF01_AE, CRF_BC, B’ and B strains V3 loop were 3.29±0.97(n=227), 3.11±0.42(n=55), 3.73±0.93(n=49) and 3.50±0.97(n=10) respectively. Conclusion The main types of V3 loop central motif were GPGQ and GPGR in Shenzhen. Co-receptor usage and phenotype prediction results showed that the HIV-1 epidemic in Shenzhen is still slow replication of non-syncytium-inducing virus.