哈尔滨市新确诊HIV-1 CRF01__AE亚型nef序列分析  

Variability of nef sequences from newly diagnosed HIV-1 CRF01__AE infections in Harbin city of China

作　　者：曹立君[1] 李庆海[2] 杨全[2] 岳超[2] 彭鑫[2] 杨坤[2] 唐煜辉[3] 王福祥[1] CAO Li-jun LI Qing-hai YANG Quan YUE Chao PENG Xin YANG Kun TANG Yu-hui WANG Fu-xiang(Department of Infectious Diseases, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150081 , China)

机构地区：[1]哈尔滨医科大学附属第四临床医学院感染科,哈尔滨150001 [2]哈尔滨医科大学药学院系统组学中心哈尔滨医科大学基因组中心,哈尔滨150081 [3]大庆市疾病预防控制中心,黑龙江大庆163001

出　　处：《中国病毒病杂志》2016年第1期34-39,共6页Chinese Journal of Viral Diseases

基　　金：黑龙江省自然科学基金(H201383);黑龙江省大学生创业创新基金(201510226017;201410226040)

摘　　要：目的分析哈尔滨市新确诊人类免疫缺陷病毒1型(HIV-1)CRF01_AE亚型nef基因及nef蛋白重要功能区氨基酸的变异性。方法从56例2014-2015年新确诊感染者外周血单核细胞(peripheral blood mononuclear cell,PBMC)中提取基因组DNA,扩增全长nef基因,并测序。构建nef基因系统发育树,分析基因特点;将nef氨基酸序列与中国和国际CRF01_AE共享序列进行比对,分析nef蛋白重要功能区氨基酸的变异性。结果 56株nef基因序列与不同CRF01_AE亚簇聚集,其中26株与CRF01-4亚簇聚集,24株与CRF01-5亚簇聚集,6株与CRF01-1亚簇聚集。亚簇内基因距离分析发现,CRF01-5亚簇最小,CRF01-4亚簇次之,CRF01-1亚簇最大。nef蛋白多个功能区氨基酸序列高度保守,但长度可变区、凋亡结构域、酸性区和C端PAK结合区变异率较高,其中凋亡区M50I突变和PAK结合区K192R突变多见于CD4数较低的样本(P<0.05)。结论哈尔滨市新确诊CRF01_AE亚型nef基因的组成较为复杂,可能来源于多个地区和多种感染途径;凋亡结构域和PAK结合区氨基酸变异与疾病进展相关。Objective To analyze the nefgene and deducted amino acid sequences and the variability of functional nef domains in newly diagnosed HIV-1-infected individuals in Harbin city of China. Methods Genomic DNAs were extracted from peripheral blood mononuclear cells（PBMCs）of 56HIV-1-infected individuals who were diagnosed as HIV-1positive from 2014 to 2015.The full-length of nefgene was amplified and subjected to sequencing.Phylogenetic tree was constructed to analyze nefgene feature;and deducted nef amino acid sequences were aligned with consensus references of China and worldwide CRF01_strains to analyze the variability of the main functional domains of the nef proteins. Results The 56 nefgene sequences clustered with three CRF01_sub-lineages as follows：26sequences clustered with CRF01-4,24 with CRF01-5,and 6with CRF01-1.Intra-sublineage genetic distance analysis revealed the smallest sequences in CRF01-5and longest distances in CRF01-1.The amino acid sequences of multiple functional nef domains were highly conserved,but the lengths of the variable regions,apoptosis motifs,acid regions and C-terminal PAK binding domains were highly variable with multiple site mutations.The M50 Imutation within the apoptosis motif and the K192 Rmutation within C-terminal PAK binding domain were frequently observed in samples with CD4 counts fewer than 200. Conclusions The genetic feature of CRF01_nefgenes from newly diagnosed HIV-1-infected individuals is complicated,suggesting that current epidemic of CRF01_viruses in Harbin city of China might be originated from different geographical regions of China and through multiple infection routes.The nef amino acid mutations M501 and K192Rin the apoptosis motif and the C-terminal PAK binding domain,respectively,were associated with disease progress.

关 键 词：人类免疫缺陷病毒1型(HIV-1) CRF01__AE nef基因序列 功能区 

分 类 号：R512.91[医药卫生—内科学]