CAJ | 학술논문

目的调查同一供体来源的人类免疫缺陷病毒(HIV)-1感染不同个体后病毒包膜糖蛋白的变异、病毒侵入靶细胞能力以及包膜抗原主要中和表位的变化,为了解病毒感染规律及机体抗病毒免疫奠定基础.方法对病毒包膜糖蛋白基因序列进行基因离散分析;用包膜蛋白表达质粒与HIV-1骨架质粒共转染293T细胞构建包膜包膜假病毒,用假病毒感染HIV-1靶细胞U87.CD4.CCR5或U87.CD4.CXCR4细胞检测假病毒侵入靶细胞的能力及病毒亲嗜性;对包膜糖蛋白中已知的广谱中和抗体识别表位进行分析.结果 24个有完整开放读码框的env基因克隆与河南省HIV-1毒株CNHN24的基因离散率为(7.91±0.78)%,与云南省分离毒株RL42的基因离散率为(6.90± 40.79)%.各可变区基因离散率呈现严重不均衡性,其中,V1/V2区的离散率最高,V4区的离散率次之,V3区离散率最小.包膜假病毒中既有CCR5亲嗜性和CXCR4亲嗜性的,也有双亲嗜性的包膜.而且上述包膜中主要中和表位IgG1b12、2F5和4E10抗体识别表位保守,但447-52D抗体识别表位变异较大.结论同一来源的HIV包膜糖蛋白在4～7年间的不同受者体内发生了较大变异并影响了病毒侵入靶细胞的能力;主要广谱中和抗体的识别表位部分保守.
목적 조사동일공체래원적인류면역결함병독(HIV)-1감염불동개체후병독포막당단백적변이、병독침입파세포능력이급포막항원주요중화표위적변화,위료해병독감염규률급궤체항병독면역전정기출.방법 대병독포막당단백기인서렬진행기인리산분석;용포막단백표체질립여HIV-1골가질립공전염293T세포구건포막포막가병독,용가병독감염HIV-1파세포U87.CD4.CCR5혹U87.CD4.CXCR4세포검측가병독침입파세포적능력급병독친기성;대포막당단백중이지적엄보중화항체식별표위진행분석.결과 24개유완정개방독마광적env기인극륭여하남성HIV-1독주CNHN24적기인리산솔위(7.91±0.78)%,여운남성분리독주RL42적기인리산솔위(6.90± 40.79)%.각가변구기인리산솔정현엄중불균형성,기중,V1/V2구적리산솔최고,V4구적리산솔차지,V3구리산솔최소.포막가병독중기유CCR5친기성화CXCR4친기성적,야유쌍친기성적포막.이차상술포막중주요중화표위IgG1b12、2F5화4E10항체식별표위보수,단447-52D항체식별표위변이교대.결론 동일래원적HIV포막당단백재4～7년간적불동수자체내발생료교대변이병영향료병독침입파세포적능력;주요엄보중화항체적식별표위부분보수.
Objective To investigate the evolution of HIV-1 envelope (env) gene from the individuals infected by the virus from one donor, the entry mediated by the envelope glycoprotein and the variation in the main neutralizing epitopes of envelope. Methods The genetic distances of the HIV-1 envelope genes derived from previous studies were analyzed. A series of envelope-pseudotyped viruses were constructed by co-transfecting HEK293T cells with a HIV-1 plasmid bearing the firefly luciferase reporter gene and an envelope expression plasmid. The entry ability of the envelope-pseudotyped viruses into U87. CD4. CCR5 or U87. CD4. CXCR4 cell lines was examined. The ami-no acid sequences representing the epitopes to the broad-neutralizing antibodies within the envelope glycoproteins were also investigated. Results It was found that the genetic distance of the 24 env genes with complete open reading frame was (7.91 ±0.78)% towards HIV-1 CNHN24, and (6.90 ±0.79)% towards RL42. Among the variable regions, the genetic distance of V1/V2 showed the biggest distance, and that of V3 showed the smallest distance. There were CCR5-tropic, CXCR4-tropic and CCR5/CXCR4-dual-tropic Env-pseudoviruses. Furthermore, in these envelopes, the epitopes to IgG1 b12 2F5 and 4E10 antibody were conserved, while the epitope to 447-52D was variable. Conclusion There is definite env gene variation among the viruses derived from the same donor. The variation influences the entry ability and tropism of emelope pseudoviruses. The epitopes to the main broad-neutralizing antibodies are conserved.