摘要
目的分析我国HIV-1B′亚型R5或R5/X4嗜性毒株在GHOST细胞的感染性。方法采用传统的共培养方法从HIV-1感染者PBMC中分离并培养病毒,用表达CD4和趋化因子受体CCR5或CXCR4的GHOST细胞系,测定毒株的辅助受体利用情况,使用相同病毒量即2ng的HIV-1 p24分别感染表达不同受体的GHOST细胞系,通过流式细胞仪检测分析绿色荧光蛋白(GFP)的表达反映病毒感染细胞的能力。结果35例B′亚型毒株利用CCR5受体,占22例(62.85%),双嗜性即CCR5/CX- CR4均阳性占13例(37.15%)。GHOST-R5/X4细胞的感染性分析结果显示,RS/X4双嗜性毒株的感染性明显高于CD4>200/μl的R5毒株的感染性(P<0.05);R5/X4毒株与CD4≤200/μl的R5毒株感染性比较差异无统计学意义(P>0.05);CD4>200/μl的R5毒株与CD4≤200/μl的R5毒株感染性比较差异有统计学意义(P<0.05);GHOST-CCR5细胞感染性分析结果显示:R5/X4双嗜性毒株的感染性明显下降,与CD4>200/μl的R5毒株的感染性比较差异无统计学意义(P>0.05)。利用相同剂量的双嗜性毒株分别感染R5、X4或R5/X4的GHOST细胞系,显示双嗜性毒株可同时利用CCR5和CXCR4辅助受体,但69.23%的R5/X4毒株以CCR5受体为主,30.77%的R5/X4毒株以CXCR4受体为主。结论HIV-1 B′亚型R5/X4病毒不仅有更广泛的宿主细胞嗜性,而且在GHOST-R5/X4细胞中感染性明显提高。持续使用CCR5受体的毒株在疾病进展的过程中虽然辅助受体的利用是一样的,但病毒感染细胞的能力增加。
Objective To analyze the infectivity of HIV-1B′ subtype R5-tropic or R5/X4-dual tropic isolates in GHOST cell line. Methods The clinic viruses were isolated from PBMC of HIV-1 infectioners by coculture. Meanwhile, their co-receptor usages were determined with GHOST （3） cell fines, stably expressing CD4 and the chemokine receptor CCR5, CXCR4 and R5/X4. Infectivity of 2 ng p24 antigen equivalent virus to different GHOST cell lines was determined using flow cytometer. Results 62.85%（22/35）HIV-1B′ strains used CCR5 as co-receptor; 37.15% （13/35） isolates used R5/X4 as co-receptor. The infectivity of R5/X4 dualtropic viruses was higher than one of R5 tropic viruses from CD4 〉 200/μl individuals （ P 〈 0.05） and the infectivity of R5 strains from CD4≤200/μl individuals was higher than R5 strains from CD4〉 200/μl （P 〈 0.05） in GHOST-R5/X4 cell line. There is no significant difference of viral infectivity between R5/X4 tropism and R5 tropism from CD4≤200/ μl individuals（ P 〉 0.05）in GHOST-CCR5 cell. Using 2 ng p24 antigen equivalent virus to infect three kinds GHOST cell lines expressing CCRS, CXCR4 and R5/X4 co-receptor respectively, 69.23% dual-tropic viruses used mainly CCR5 co-receptor. 30.77% dual-tropic viruses used mainly CXCR4 co-receptor. Conclusion R5/ X4 strains not only have abroad host cell but also have high infectivity in GHOST-R5/X4 cell. Persistent CCR5 utihzation isolates have different infectivity.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2007年第12期1098-1101,共4页
Chinese Journal of Microbiology and Immunology
基金
国家重点基础研究(973)发展计划(2005CB523103
2005CB522903)
CIPRA项目(U19AIS1915-03)
