Routledge E G, McQuillin J, Samson A C, Toms G L
J Med Virol. 1985 Mar;15(3):305-20. doi: 10.1002/jmv.1890150311.
Twelve clones of murine hybridoma cells secreting antibody specific for respiratory syncytial (RS) virus were classified into four groups on the basis of their pattern of staining of unfixed RS virus-infected HEp-2 cells in an indirect immunofluorescence test. Three of the groups reacted with virus antigens present on the membrane of the cells, whilst the fourth group failed to stain most live cells, suggesting specificity for an antigen expressed internally. Representative monoclonals from the membrane antigen staining groups immunoprecipitated the 86K glycoprotein (G), 50K plus 19K glycoprotein (F1,2) and a 23K non-glycosylated protein (VP23). A representative monoclonal from the fourth group that appeared to stain an internally expressed protein immunoprecipitated the virion 34K phospho-protein (P). All four monoclonals stained acetone-fixed tissue culture cells infected with either the Long strain of RS virus or with strains isolated in Newcastle during the 1965, 1972, and 1983 winter epidemics. The anti-fusion protein antibody stained acetone-fixed cells from all of 26 nasopharyngeal secretions from infants with RS virus infection. The anti-G glycoprotein antibody and the anti-VP23 antibody stained cells from secretions poorly or not at all, whilst the anti-P protein antibody stained cells in half the secretions tested but reacted with only a small proportion of cells in comparison with the anti-F or polyclonal antibodies. A pool of all four monoclonals produced more intense staining than the anti-F monoclonal alone and gave a more clearly defined staining reaction than the polyclonal antiserum used for routine diagnosis in over half the secretions. These results indicate that monoclonal antibodies will be of value in the diagnosis of RS virus by indirect immunofluorescence if care is taken in the selection of a suitable pool.
在间接免疫荧光试验中，根据未固定的呼吸道合胞（RS）病毒感染的HEp-2细胞的染色模式，将12个分泌针对RS病毒的特异性抗体的鼠杂交瘤细胞克隆分为四组。其中三组与细胞表面存在的病毒抗原发生反应，而第四组未能对大多数活细胞进行染色，这表明其对细胞内表达的抗原具有特异性。来自膜抗原染色组的代表性单克隆抗体免疫沉淀了86K糖蛋白（G）、50K加19K糖蛋白（F1,2）和一种23K非糖基化蛋白（VP23）。来自第四组的一个代表性单克隆抗体似乎对细胞内表达的一种蛋白进行了染色，该抗体免疫沉淀了病毒粒子的34K磷蛋白（P）。所有这四种单克隆抗体均能对感染RS病毒Long株或在1965年、1972年和1983年冬季流行期间于纽卡斯尔分离出的毒株的丙酮固定的组织培养细胞进行染色。抗融合蛋白抗体对来自26例RS病毒感染婴儿的所有鼻咽分泌物的丙酮固定细胞进行了染色。抗G糖蛋白抗体和抗VP23抗体对分泌物中的细胞染色不佳或根本不染色，而抗P蛋白抗体对一半的受试分泌物中的细胞进行了染色，但与抗F单克隆抗体或多克隆抗体相比，仅与一小部分细胞发生反应。由所有这四种单克隆抗体组成的混合抗体产生的染色比单独的抗F单克隆抗体更强，并且在超过一半的分泌物中，其染色反应比用于常规诊断的多克隆抗血清更清晰明确。这些结果表明，如果在选择合适的混合抗体时加以注意，单克隆抗体在通过间接免疫荧光诊断RS病毒方面将具有价值。
