Pande H, Campo K, Shanley J D, Creeger E S, Artishevsky A, Gallez-Hawkins G, Zaia J A
Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California.
J Gen Virol. 1991 Jun;72 ( Pt 6):1421-7. doi: 10.1099/0022-1317-72-6-1421.
We have developed a hybridoma, designated 25G11, which produced a monoclonal antibody (MAb) reactive with a 52K protein of murine cytomegalovirus (MCMV). This MAb, 25G11, was reactive with a protein band of 52K in MCMV-infected cell lysates and with a protein of 49K in human CMV (HCMV)-infected cell lysates as detected by immunoblot analysis. With purified MCMV virions, 25G11 gave a faintly immunoreactive band of 52K. However, no immunoreactive protein band was detected with purified HCMV virions, nor with purified HCMV or MCMV envelope preparations. By immunocytochemistry, 25G11 detected viral antigen primarily in the nucleus of HCMV- or MCMV-infected cells. The antibody 25G11 was used to screen a lambda gt11 library of HCMV DNA fragments. One of the isolated clones (lambda 32323B) was employed for gene mapping on the HCMV genome, which suggested that the immunoreactive HCMV protein was the DNA-binding protein (ICP36). Analysis of the recombinant fusion protein with antibody 25G11 and with an MAb (CH16) specific for an HCMV DNA-binding protein confirmed the identity of the cross-reacting protein as ICP36. Furthermore, we found that whereas the epitope recognized by 25G11 was conserved between HCMV and MCMV proteins, the epitope recognized by CH16 was unique to HCMV and thus represents a variable region in the protein.
我们培育出了一种名为25G11的杂交瘤，它能产生一种与鼠巨细胞病毒（MCMV）的52K蛋白发生反应的单克隆抗体（MAb）。通过免疫印迹分析检测到，这种单克隆抗体25G11能与MCMV感染的细胞裂解物中的一条52K蛋白带发生反应，也能与人类巨细胞病毒（HCMV）感染的细胞裂解物中的一种49K蛋白发生反应。对于纯化的MCMV病毒粒子，25G11产生了一条微弱反应的52K免疫反应带。然而，无论是纯化的HCMV病毒粒子，还是纯化的HCMV或MCMV包膜制剂，均未检测到免疫反应蛋白带。通过免疫细胞化学方法，25G11主要在HCMV或MCMV感染细胞的细胞核中检测到病毒抗原。抗体25G11被用于筛选HCMV DNA片段的λgt11文库。其中一个分离的克隆（λ32323B）被用于HCMV基因组上的基因定位，这表明具有免疫反应性的HCMV蛋白是DNA结合蛋白（ICP36）。用抗体25G11和一种对HCMV DNA结合蛋白具有特异性的单克隆抗体（CH16）对重组融合蛋白进行分析，证实了交叉反应蛋白为ICP36。此外，我们发现，虽然25G11识别的表位在HCMV和MCMV蛋白之间是保守的，但CH16识别的表位是HCMV特有的，因此代表了该蛋白中的一个可变区域。
