【摘要】 采用同源重组技术构建了非洲猪瘟病毒(African swine fever virus, ASFV)MGF110-3L基因缺失(ASFV-△MGF110-3L)重组病毒，通过检测该重组病毒在原代猪肺泡巨噬细胞(porcine alveolar macrophages, PAMs)中的复制水平，探究其在ASFV复制周期中的功能。结果显示，MGF110-3L基因缺失重组病毒的复制动力学与其亲本病毒ASFV-WT无差异，表明缺失MGF110-3L基因不影响病毒复制。进一步将ASFV-WT和ASFV-△MGF110-3L分别感染PAMs,并于感染后4、12、20 h收集样品，进行转录组测序(RNA-seq),发现缺失MGF110-3L显著促进IFN-β、CCL2、CCL4等细胞因子基因的转录；KEGG通路分析显示，与ASFV-WT感染组相比，ASFV-△MGF110-3L感染组中差异表达基因主要与细胞因子-细胞因子受体之间的相互作用相关。本研究通过构建ASFV-△MGF110-3L缺失毒，并对其与野毒进行转录组分析发现，MGF110-3L基因作为ASFV的复制非必需基因具有拮抗细胞因子表达的功能，结果不仅为阐明MGF110的生物学功能提供科学数据，还为深入揭示ASFV拮抗宿主细胞天然免疫反应的分子机制提供了参考。更多还原

【Abstract】 Using homologous recombination technology,an MGF110-3L gene-deleted recombinant virus(ASFV-△MGF110-3L) was constructed and used to explore its role in the African swine fever virus(ASFV) replication cycle by evaluating the replication efficiency of the recombinant virus in primary porcine alveolar macrophages(PAMs).The results showed that the replication kinetics of ASFV-△MGF110-3L was comparable to that of ASFV-WT,indicating that MGF110-3L deletion does not affect viral replication.To further elucidate the biological function of the MGF110-3L protein in the ASFV replication cycle,PAMs were infected with ASFV-WT or ASFV-△MGF110-3L and samples were collected at 4,12,and 20 hours post-infection for transcriptomic sequencing(RNA-seq) analysis.The findings revealed that the deletion of MGF110-3L significantly promoted the transcription of IFN-β,CCL2,and CCL4 and other cytokine genes.KEGG pathway analysis indicated that the differentially expressed genes in the ASFV-△MGF110-3L-infected group were primarily associated with the cytokine-cytokine receptor interactions compared with the ASFVWT infected group.In summary,ASFV-△MGF110-3L recombinant virus was generated and used to investigate the functions of MGF110-3L gene.The findings demonstrated that MGF110-3L is a non-essential gene for ASFV replication,and its functions to antagonize cytokine expression,which provides valuable scientific data for understanding the biological function of MGF110 and offers insights into the molecular mechanisms by which ASFV counteracts the host’s innate immune response.更多还原