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目的研究SHIV-XJ02170在中国恒河猴感染后期传代过程中病毒和宿主的变化特点,并分析env基因的序列变异。方法将感染中国恒河猴G0401V后期(5年)的SHIV-XJ02170病毒垂直传代2只猴(G0401V→G0402V→0403V),同时,剔除G0401V猴CD8+T细胞使潜伏的病毒大量复制后传代1只猴(G0401V→G0404V),应用流式细胞术、病毒载量测定、序列分析等方法研究该病毒在猴体内长期适应后的病毒和免疫学指标及序列变异特点。结果 G0401V在感染后期仍能稳定传代,且表现出毒力增强的特点。其传代猴G0402V在传代后41 d死亡,外周血CD4+T淋巴细胞衰竭,仅为43个/mL,符合艾滋病感染猴快速进展型的特征。剔除体内CD8+T细胞之后的传代猴G0404V的表现类似G0401V,即长期低水平的病毒血症水平。env基因序列分析发现SHIV-XJ02170在G0401V体内长期适应后发生了可遗传的序列变异,并引起糖基化位点的改变。结论 SHIV-XJ02170在猴体长期适应后的传代过程中表现出向强毒株过渡的特征,为进行SHIV-XJ02170感染性克隆的构建奠定了良好的实验基础。
Objective To investigate the variation of SHIV-XJ02170 virus and host during the late passage of rhesus monkey infection in China and to analyze the sequence variation of env gene. Methods Two monkeys (G0401V → G0402V → 0403V) were passaged vertically with SHIV-XJ02170 virus infected with G0401V of Chinese rhesus at the later stage (5 years). Meanwhile, a large number of latent viruses were replicated and 1 passaged after passage of G0401V monkey CD8 + T cells (G0401V → G0404V). The virus and immunological indexes and sequence variations of the virus after long-term adaptation in the monkey were studied by flow cytometry, viral load assay and sequence analysis. Results G0401V was stable in the later stage of infection and showed enhanced virulence. Its passage monkey G0402V died at 41 days after passage, CD4 + T lymphocyte depletion in peripheral blood was only 43 cells / mL, which was in line with the rapid progress of AIDS-infected monkeys. The passage monkey, G0404V, which excludes CD8 + T cells in vivo, behaves like G0401V, a long term, low level of viremia. Env gene sequence analysis found that SHIV-XJ02170 in G0401V long-term adaptation occurred after the genetic sequence variation, and cause changes in glycosylation sites. Conclusion SHIV-XJ02170 showed the transition to virulent strains during the passage of long-term adaptation of SH-XJ02170, which laid a good experimental foundation for the construction of SHIV-XJ02170 infectious clone.