Serial Passage of Virus Clade C SHIVCHN19P4 in Chinese Rhesus Macaques

Abstract

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. 目的研究C亞型SHIVCHN19P4強毒株在中國恒河猴體內傳代中病毒學和免疫學等反應的變化特點,分離制備SHIVCHN19P4中國恒河猴傳代適應株病毒。方法選擇4只健康成年恒河猴,其中兩只經后肢靜脈感染SHIVCHN19P4病毒,60 d后,分別采集EDTA抗凝全血靜脈途徑傳代至另兩只猴,使用流式細胞術、PCR、結合抗體檢測和序列分析等方法研究傳代動物病毒學、免疫學和序列變異特點。選擇性從傳代動物感染急性期外周血中分離PBMC,CD8+T細胞敲除后與正常PBMC共培養分離病毒。結果 4只傳代動物均獲得系統性感染,且傳代后病毒毒力明顯增強,序列分析發現SHIVCHN19P4病毒序列在傳代過程中發生適應性改變;同時,成功分離制備SHIVCHN19P4傳代適應性病毒株。結論 SHIVCHN19P4在中國恒河猴體內適應性傳代研究為進一步建立C亞型SHIV強毒株/SAIDS模型奠定了良好的實驗基礎,為研究C亞型HIV-1流行株致病特點以及預防性黏膜疫苗和殺微生物的有效性評價提供了數據支持。 更多還原