Human cytomegalovirus (HCMV) is a paradigm for pathogen-mediated immune evasion. The immune response to HCMV has been intensively studied for many years and still remains the focus of attention for numerous research groups. UL23 is an early gene of HCMV, belonging to the US22 gene family, encoding protein UL23. However, no monoclonal antibodies against to HCMV UL23 protein have been reported to prepare for the research. In this study, we prepared a highly specific monoclonal antibody against UL23 protein by alternately immunizing BALB/C mice with both UL23 recombinant protein and HCMV Towne. Recombinant protein UL23 was used as a detection antigen to screen 305 strains of hybridoma cells. One of them was identified to secrete IgG1 mAb named as 26C5. Western blotting results showed that not only the overexpressed UL23 protein in 293T cells but also the viral UL23 protein in HCMV-infected human foreskin fibroblast cells specifically were recognized by 26C5 mAb. Notably, we found that UL23 protein were enriched by 26C5 mAb in coimmunoprecipitation experiment with high potency and the native form of UL23 protein localizing primarily in the cytoplasm were recognized by 26C5 mAb in immunofluorescence assay with high specificity. The monoclonal antibody obtained in this study lays the foundation for further study of HCMV UL23 protein.

中文翻译：

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抗人巨细胞病毒UL23的小鼠单克隆抗体的产生和应用。

人类巨细胞病毒（HCMV）是病原体介导的免疫逃逸的范例。对HCMV的免疫反应已经进行了多年深入研究，仍然是许多研究小组关注的焦点。UL23是HCMV的早期基因，属于US22基因家族，编码蛋白UL23。然而，没有针对HCMV UL23蛋白的单克隆抗体的报道为该研究做准备。在这项研究中，我们通过用UL23重组蛋白和HCMV Towne交替免疫BALB / C小鼠，制备了针对UL23蛋白的高度特异性单克隆抗体。重组蛋白UL23被用作检测抗原，以筛选305个杂交瘤细胞株。鉴定出其中之一可分泌称为26C5的IgG1 mAb。Western印迹结果表明，不仅26C5 mAb可特异性识别在293T细胞中过表达的UL23蛋白，而且在HCMV感染的人包皮成纤维细胞中也可特异性识别病毒UL23蛋白。值得注意的是 我们发现，在高效共免疫沉淀实验中，26C5 mAb富集了UL23蛋白，在免疫荧光测定中，26C5 mAb可以识别主要位于细胞质中的天然形式的UL23蛋白。本研究获得的单克隆抗体为进一步研究HCMV UL23蛋白奠定了基础。