Zal B, Booth J, Chadwick J, Baboonian C
Department of Cardiological Sciences, St. George's Hospital Medical School, London, England.
J Med Virol. 1999 Mar;57(3):290-7.
Of nine mouse monoclonal antibodies (MAbs) directed against the lower matrix protein (pp65; ppUL83) of human cytomegalovirus (HCMV), all immunoprecipitated the 65-kDa protein. Only five were reactive by Western blotting, however, and four of these mapped to linear antigenic epitopes located between amino acids 184-195 (MAb C6), 343-357 (MAb C11), 448-462 (MAb C5), and 448-459 (MAb C13). The epitope specificity of the fifth antibody (MAb C3) and the four that recognised nonlinear sites could not be determined. Competition binding studies using HCMV antigen extracted from productively infected human embryonic lung fibroblasts (HELF), in an enzyme immunoassay (EIA), showed that three of the antibodies reactive with linear epitopes and two of those reactive with conformational epitopes (MAbs C3, C6, C11, C14, and C18), were unique in their binding specificities. MAb C4 competed with MAb C8 and MAb C5 competed with MAb C13 for binding to ppUL83. One of the linear epitopes identified, corresponding to amino acids SAFVFPTKDVAL (MAb C6), was an epitope described previously for CD8+ cytotoxic T lymphocytes.
针对人巨细胞病毒（HCMV）下层基质蛋白（pp65；ppUL83）的九种小鼠单克隆抗体（MAb）均能免疫沉淀65 kDa的蛋白。然而，通过蛋白质印迹法只有五种具有反应性，其中四种定位于位于氨基酸184 - 195（单克隆抗体C6）、343 - 357（单克隆抗体C11）、448 - 462（单克隆抗体C5）和448 - 459（单克隆抗体C13）之间的线性抗原表位。第五种抗体（单克隆抗体C3）以及识别非线性位点的四种抗体的表位特异性无法确定。在酶免疫测定（EIA）中，使用从高效感染的人胚肺成纤维细胞（HELF）中提取的HCMV抗原进行竞争结合研究，结果表明，三种与线性表位反应的抗体以及两种与构象表位反应的抗体（单克隆抗体C3、C6、C11、C14和C18）在结合特异性方面是独特的。单克隆抗体C4与单克隆抗体C8竞争，单克隆抗体C5与单克隆抗体C13竞争与ppUL83的结合。所鉴定的一个线性表位，对应于氨基酸SAFVFPTKDVAL（单克隆抗体C6），是先前针对CD8 + 细胞毒性T淋巴细胞描述的表位。
