摘要
目的 探讨人巨细胞病毒 (HCMV)主要基因序列表达情况对其感染结局的影响。方法采用不同滴度HCMV感染传代培养的人胚肺成纤维细胞 ,建立HCMV梯度感染细胞模型。分别应用FQ PCR法测定感染细胞内IE基因拷贝数、免疫组化法联合计算机病理图文分析系统检测p5 2蛋白表达水平以及RT PCR法测定MCPmRNA转录水平 ,光镜观察致细胞病变作用 (CPE)、电镜检查细胞超微结构改变。结果 IE基因在病毒滴度最低的A组细胞内负荷量最低 (P <0 .0 5 ,P <0 .0 1) ;p5 2蛋白在病毒滴度较高的B、C组表达水平明显高于A组 (P <0 .0 1) ,其表达定位于受染细胞核内 ;仅在B、C组内检测到MCPmRNA。A组未检出细胞病变 ,B、C组细胞检测到随感染时间延长而加重的CPE及超微结构改变。结论 HCMV受染细胞内IE基因负荷量与病毒基因组表达启动密切相关 ,p5 2蛋白高效表达是病毒基因组完整表达的必要条件 ,MCPmRNA转录是活动性HCMV感染的标志。
Objective To investigate the relationship between the expression of viral main genes and pathologic outcomes of infected cells in HCMV infection. Methods HCMV echelon-infected cell model was set up in vitro by coincubating passage cultured cells and HCMV with different titers. FQ-PCR was performed to evaluate the number of IE gene copies in infected cells and the expression of p52 was determined by immunohistochemical test and quantitated by computer imaging analysis. Major capsid protein (MCP) mRNA was also measured by RT-PCR. Meanwhile the cytopathologic effects(CPE) was observed by microscopy and TEM. Results Compared with group B and C in high HCMV titer, group A with low viral titer showed low IE gene load(P<0.05, P<0.01), while in group B and C, p52 expression was significantly elevated (P<0.01) and MCP mRNA was detected. At the same time, group B and C carried developing CPE and ultrastructural impairment during HCMV infection, that wasn′t detected in group A. Conclusion In HCMV infected cells, IE gene load is associated with initiation of viral genome functioning and efficient p52 expression is necessary for its complete performance. The detected MCP mRNA is an indicator of active HCMV infection.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2004年第10期834-838,共5页
Chinese Journal of Microbiology and Immunology
