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目的 探讨宿主因子衣被蛋白复合物亚基β2(Coatomer subunit beta 2, COPB2)在痘苗病毒（Vaccinia virus,VACV）复制过程中的作用及其作为广谱抗正痘病毒药物靶点的潜力。方法 整合分析现有高通量宿主因子筛选研究，挖掘高频出现的促病毒宿主因子，选定COPB2为研究对象，以已知宿主因子整合素亚基β1(Integrin subunit beta 1, ITGB1)为阳性对照。在HeLa和HEK-293T细胞中构建siRNA介导的基因敲低模型，通过CCK-8法评估细胞活性，利用噬斑形成实验检测子代病毒滴度，使用qPCR检测病毒基因组复制载量，并通过Western blot检测病毒早期与晚期蛋白表达水平。结果 成功构建基于siRNA的COPB2与ITGB1基因敲低模型。与对照组相比，敲低COPB2或ITGB1均可显著抑制VACV复制，表现为子代病毒滴度明显降低，病毒基因组复制载量下降以及病毒早期及晚期蛋白表达减少。结论 COPB2是促进VACV高效复制的重要宿主因子，初步推测其功能可能主要参与病毒生命周期的中晚期阶段，但其具体作用环节及分子机制仍有待进一步系统阐明。本研究提示，COPB2具有成为天花、猴痘等正痘病毒广谱宿主靶向抗病毒干预靶点的潜在价值。
Abstract：Objective To investigate the role of the host factor COPB2 in the replication of vaccinia virus(VACV) and to evaluate its potential as a broad-spectrum anti-orthopoxvirus drug target. Methods We performed an integrated analysis of existing high-throughput host factor screening studies to identify frequently implicated proviral host factors. COPB2 was selected as the primary research subject, with the known host factor ITGB1 serving as a positive control. Gene knockdown models for COPB2 and ITGB1 were established in HeLa and HEK-293T cells using siRNA-mediated gene knockdown. Cell viability was assessed by CCK-8 assay. The viral titer of progeny viruses was determined by plaque formation assay. Viral genomic replication load was measured by qPCR. The expression levels of viral early and late proteins were detected by Western blot. Results SiRNA-mediated knockdown models for COPB2 and ITGB1 were successfully constructed. Compared to the control group, knockdown of either COPB2 or ITGB1 significantly inhibited VACV replication. This inhibition was evidenced by a marked reduction in progeny viral titer, a decrease in viral genome replication load, and diminished expression of viral early and late proteins.Conclusion COPB2 is an important host factor that promotes efficient VACV replication. Preliminary evidence suggests its function is likely involved in the mid-to-late stages of the viral life cycle, although the specific molecular mechanisms and precise steps require further systematic elucidation. This study indicates that COPB2 has potential as a hostdirected target for developing broad-spectrum antiviral interventions against orthopoxviruses, including smallpox and mpox.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.260053
中图分类号:R373
引用信息:
[1]李兆卿,霍恕婷,李若瑞,等.促病毒宿主因子COPI成员COPB2调控痘苗病毒生命周期的体外实验研究[J].病毒学报,2026,42(02):454-464.DOI:10.13242/j.cnki.bingduxuebao.260053.
基金信息:
中国疾病预防控制中心青年科学基金项目（项目号：2024A103）,题目：基因组大片段缺失猴痘病毒新变异株的生物学功能改变及其机制研究~~
2026-02-13
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