Massicotte H, Harley J B, Bell D A
University of Western Ontario, Department of Medicine, University Hospital, London, Canada.
J Autoimmun. 1992 Dec;5(6):771-85. doi: 10.1016/0896-8411(92)90192-s.
Human-human hybridomas obtained from the separate fusion of tonsillar lymphoid cells from three different normal individuals to the lymphoblastoid cell line GM 4672 were screened by ELISA for the presence of autoantibody to Ro(SS-A). Those anti-Ro(SS-A) reactive hybridomas were then cloned by limiting dilution. Nineteen monoclonal IgM anti-Ro(SS-A) antibodies were obtained, which showed specificity to Ro(SS-A) by ELISA and Western blotting (60 kDa). Some of these monoclonal anti-Ro(SS-A) antibodies showed reactivity to DNA (2/19), cardiolipin (9/19), Sm/RNP (15/19) by ELISA, and to IgG (12/19) and La(SS-B) (19/19) by ELISA and Western blotting. None showed reactivity to the unrelated proteins casein and BSA, nor to RNA. Inhibition studies revealed that the binding to Ro(SS-A) of both IgM hybridoma monoclonal and SLE serum polyclonal IgM anti-Ro(SS-A) antibodies was inhibited with Ro(SS-A), La(SS-B) and Sm/RNP but not with IgG, DNA, RNA and BSA. These data indicate that (1) normal humans have the genetic potential to express antibodies to Ro(SS-A) and (2) the normally derived monoclonal and SLE serum IgM anti-Ro(SS-A) antibodies share similar antigen binding properties and therefore may possibly originate from a common pool of precursor B cells.
将来自三名不同正常个体的扁桃体淋巴样细胞分别与淋巴母细胞系GM 4672融合得到人-人杂交瘤,通过酶联免疫吸附测定(ELISA)筛选其中是否存在抗Ro(SS - A)自身抗体。然后通过有限稀释法克隆那些抗Ro(SS - A)反应性杂交瘤。获得了19种单克隆IgM抗Ro(SS - A)抗体,通过ELISA和蛋白质印迹法(60 kDa)显示出对Ro(SS - A)的特异性。其中一些单克隆抗Ro(SS - A)抗体通过ELISA显示出对DNA(2/19)、心磷脂(9/19)、Sm/RNP(15/19)有反应性,通过ELISA和蛋白质印迹法显示出对IgG(12/19)和La(SS - B)(19/19)有反应性。没有一种抗体对不相关蛋白质酪蛋白和牛血清白蛋白以及RNA有反应性。抑制研究表明,Ro(SS - A)、La(SS - B)和Sm/RNP可抑制IgM杂交瘤单克隆抗体和系统性红斑狼疮(SLE)血清多克隆IgM抗Ro(SS - A)抗体与Ro(SS - A)的结合,但IgG、DNA、RNA和牛血清白蛋白则不能。这些数据表明:(1)正常人类具有表达抗Ro(SS - A)抗体的遗传潜力;(2)正常来源的单克隆抗体和SLE血清IgM抗Ro(SS - A)抗体具有相似的抗原结合特性,因此可能起源于共同的前体B细胞池。