相同来源HIV-1 Env、Gag基因序列变异和宿主基因多态性与疾病进展关系的分析

Association between sequence variation of Env, Gag genes from the same source and Ⅳ-1 disease progression and host genetic poIymorphism

目的探讨HIV—1基因序列变异和宿主基因多态性与疾病进展的关系。方法PCR方法从外周血细胞中扩增HIV—1 Env、Gag区片段和测序，分析序列变异、糖基化，超突变等指标。RFLP法确定宿主基因多态性。结果未治疗组中，env基因区PCR和克隆序列平均离散率分别为0．1和0．06，差异有统计学意义，治疗组内差异没有统计学意义。V3环顶端序列在未治疗和治疗组中均以GPGQ比例最大（61．5％和39％），治疗组出现稀有多肽序列如GPGH、GQGR、GLGR、12位I／V和21位Y／H变异与疾病快速进展相关。Env区段上进展较快速组（RRP）比典型进展组（TP）的糖基化程度高（平均值分别为14．56和13．20个），差异有统计学意义。Env区段上RRP比TP组GA取代百分率和绝对数平均值都高（8．7％／6．9％和10．1／7．6），差异也有统计学意义。TP组中SDF1—3’A和CCR2V62I基因频率均高于RRP组，但差异没有统计学意义。CX3CR1 V249I／M280T与疾病快速进展没有显著相关性。结论V3区序列主要位点的氨基酸变异、Env区段糖基化程度、GA取代与疾病进展相关。SDF1—3’A、CCR2V641和CX3CR1 2491／280M与疾病进展均无显著相关性。

Objective To understand the relationship between the HIV-1 viral sequence variation and host factors associated with HIV-1 disease progression. Methods Env and gag fragments of HIV-1 were amplified with PCR, cloned and sequenced. Bioinformatics was employed to find the genetic variation, N-linked glycosylation, hypermutation etc. Host gene polymorphism was analysed by using restricted fragment length polymorphism （RFLP）. Results Significant difference was found in genetic divergence between Env PCR dominant and clonal sequences （0.1 and 0.06, respectively） in non-treated group, but no significant difference was found in the HAART treated group. V3 GPGQ accounted for the most part in both treated and nontreated groups, rare V3 loop such as GPGH, GQGR and GLGR was found in treated group, V3 substitutions of I/V （position 12） and Y/H （position 21 ） was associated with the relatively rapid progression （RRP）. Glycosylation was significantly higher in RRP than in TP for Env region, GA substitution in RRP was also significantly higher than that in TP group. SDF1- 3＇A and CCR2 V64I gene frequency was higher in TP than in RRP, but the difference was not significant. Conclusion Disease progression was associated with V3 AA change, glycosylation and GA substitution in env gene. SDFI-3＇ A, CCR2 V64I and CX3CR1 V249I/M280T was not associated with disease progression significantly.