C亚型SHIVCHN19P4强毒株在中国恒河猴体内的传代研究

Serial Passages of Virus Clade C SHIV_(CHN19P4) in Chinese Rhesus Macaques

目的研究C亚型SHIVCHN19P4强毒株在中国恒河猴体内传代中病毒学和免疫学等反应的变化特点,分离制备SHIVCHN19P4中国恒河猴传代适应株病毒。方法选择4只健康成年恒河猴,其中两只经后肢静脉感染SHIVCHN19P4病毒,60 d后,分别采集EDTA抗凝全血静脉途径传代至另两只猴,使用流式细胞术、PCR、结合抗体检测和序列分析等方法研究传代动物病毒学、免疫学和序列变异特点。选择性从传代动物感染急性期外周血中分离PBMC,CD8+T细胞敲除后与正常PBMC共培养分离病毒。结果 4只传代动物均获得系统性感染,且传代后病毒毒力明显增强,序列分析发现SHIVCHN19P4病毒序列在传代过程中发生适应性改变;同时,成功分离制备SHIVCHN19P4传代适应性病毒株。结论 SHIVCHN19P4在中国恒河猴体内适应性传代研究为进一步建立C亚型SHIV强毒株/SAIDS模型奠定了良好的实验基础,为研究C亚型HIV-1流行株致病特点以及预防性黏膜疫苗和杀微生物的有效性评价提供了数据支持。

Objective To evaluate the virological and immunological changes in rhesus macaques infected with SHIVCHN19P4 virus,isolate and prepare SHIVCHN19P4 virus stock suitable for passaging in rhesus macaques.Methods Two adult rhesus macaques were infected with SHIVCHN19P4 virus intravenously.For serial passages,10 mL of whole blood from each of the P1 rhesus macaques at 60 d was transfused intravenously into two native rhesus macaques.To analyze the changes of features in those monkeys by flow cytometry,PCR,binding antibody and sequence alignment.Simultaneously,SHIVCHN19P4 virus stock was propagated by PBMCs co-culture.Results All of the four rhesus macaques established systemic infection,the infectivity of the virus was considerably enhanced after passaging.The sequence analysis revealed that SHIVCHN19P4 acquired adaptive changes.Conclusions Our findings provide a firm basis for establishing a SHIV infection model of Chinese rhesus macaques.This model may be very useful for HIV-1 subtype C vaccine and pathogenesis studies.