Vautherot J F, Laporte J
Ann Rech Vet. 1983;14(4):437-44.
Hybrid cells secreting monoclonal antibodies against Bovine Enteric Coronavirus (BECV strain G110) were obtained by fusion between SP2/O myeloma cells and splenocytes of mice hyperimmunized with purified virus. Specificity of 8 from the 12 monoclonal antibodies was established to be anti-GP105 by immunochemical staining of viral polypeptides and by immunoprecipitation of radioactive-labelled viral proteins. The reactivity of three BECV isolates (strain G110, F15 and NCDCV) with the different monoclonal antibodies was studied by indirect immunofluorescence (IIF), ELISA, hemagglutination-inhibition and seroneutralization. G110 and F15 interacted in a similar manner with the monoclonal antibodies, but NCDCV failed to react with A9 monoclonal antibody. IIF test allowed us to detect another difference between U.K. BECV isolate and other BECV, in that the U.K. isolate did not react with F7 monoclonal antibody. No differences could be seen by IIF in the reactivity of Danish BECV isolate, Human Enteric Coronavirus (HECV), Bovine Respiratory Coronavirus (BRCV) and our BECV strains, with the 12 monoclonal antibodies. Human Respiratory Coronavirus (OC43), Porcine Hemagglutinating Encephalitis Virus (HEV) and BECV share common antigenic determinants which are outlined by the 6 non-neutralizing monoclonal antibodies. On the basis of the reactivity of coronaviruses with anti-BECV monoclonal antibodies, a distinction can be easily made between BECV and other coronaviruses, and differences were also detected within the BECV isolates. Interestingly HECV were found to be more closely related to BECV than to human OC43. These results show that monoclonal antibodies are of valuable help for biochemical and antigenic studies as well as for diagnostic procedures.
通过将SP2/O骨髓瘤细胞与用纯化病毒超免疫的小鼠脾细胞融合,获得了分泌抗牛肠道冠状病毒(BECV株G110)单克隆抗体的杂交细胞。通过对病毒多肽进行免疫化学染色以及对放射性标记的病毒蛋白进行免疫沉淀,确定了12种单克隆抗体中的8种具有抗GP105的特异性。通过间接免疫荧光法(IIF)、酶联免疫吸附测定(ELISA)、血凝抑制试验和血清中和试验,研究了三种BECV分离株(G110株、F15株和NCDCV株)与不同单克隆抗体的反应性。G110株和F15株与单克隆抗体的相互作用方式相似,但NCDCV株不与A9单克隆抗体反应。IIF试验使我们能够检测到英国BECV分离株与其他BECV之间的另一个差异,即英国分离株不与F7单克隆抗体反应。通过IIF未观察到丹麦BECV分离株、人肠道冠状病毒(HECV)、牛呼吸道冠状病毒(BRCV)和我们的BECV株与这12种单克隆抗体反应性的差异。人呼吸道冠状病毒(OC43)、猪血凝性脑脊髓炎病毒(HEV)和BECV具有共同的抗原决定簇,这由6种非中和性单克隆抗体勾勒出来。根据冠状病毒与抗BECV单克隆抗体的反应性,可以很容易地区分BECV与其他冠状病毒,并且在BECV分离株之间也检测到了差异。有趣的是,发现HECV与BECV的关系比与人OC43的关系更密切。这些结果表明,单克隆抗体对生化和抗原研究以及诊断程序有很大帮助。