Westerink M A, Campagnari A A, Wirth M A, Apicella M A
Department of Medicine, School of Medicine, State University of New York at Buffalo 14215.
Infect Immun. 1988 May;56(5):1120-7. doi: 10.1128/iai.56.5.1120-1127.1988.
A monoclonal anti-idiotypic antibody (Ab2) whose antibody combining site contained a surrogate image of the meningococcal group C capsular polysaccharide was developed. To accomplish this, a monoclonal antibody against the group C capsular polysaccharide was developed by the fusion of splenocytes from mice immunized with Neisseria meningitidis group C strain MP13 with Sp2/0-Ag14 plasmacytoma cells. Monoclonal antibody 1E4, an immunoglobulin M isotype, demonstrated binding to the serogroup C polysaccharide in enzyme-linked immunosorbent assay (ELISA). Monoclonal antibody 1E4 reacted with 30 of 30 group C strains and 1 of 36 group B strains in immunodot assay, slide agglutination, inhibition ELISA, and bactericidal assay. This monoclonal antibody was selected as idiotype (Ab1) for the development of hybridomas producing an anti-idiotype antibody. One of the hybridomas developed, designated 6F9, was capable of over 70% inhibition of 1E4 in binding in the meningococcal C polysaccharide-specific ELISA. Studies with convalescent human serum demonstrated 100% inhibition of a serogroup C-specific ELISA with 200 micrograms of 6F9 per ml and 50% inhibition of this ELISA was achieved with 50 micrograms of 6F9 per ml. Monoclonal anti-idiotype antibodies (Ab3) with specificities similar to Ab1, 1E4 were generated from BALB/c mice immunized with the Ab2 (6F9). Immunization of rabbits with 6F9 resulted in an immunoglobulin G response which was significantly greater than that of control to a titer of 1:160. These studies indicate that monoclonal 6F9 contained a surrogate image on the combining antibody site which mimicked meningococcal C polysaccharide. This surrogate image is capable of evoking antibodies to the meningococcal C polysaccharide in syngenic and xenogenic species.
一种单克隆抗独特型抗体(Ab2)被研制出来,其抗体结合位点包含脑膜炎球菌C群荚膜多糖的替代影像。为实现这一目标,通过将用脑膜炎奈瑟菌C群菌株MP13免疫的小鼠脾细胞与Sp2/0-Ag14浆细胞瘤细胞融合,研制出了一种针对C群荚膜多糖的单克隆抗体。单克隆抗体1E4,免疫球蛋白M亚型,在酶联免疫吸附测定(ELISA)中显示出与C群多糖结合。在免疫斑点测定、玻片凝集、抑制ELISA和杀菌测定中,单克隆抗体1E4与30株C群菌株中的30株以及36株B群菌株中的1株发生反应。这种单克隆抗体被选作独特型(Ab1)用于制备产生抗独特型抗体的杂交瘤。所研制出的其中一种杂交瘤,命名为6F9,在脑膜炎球菌C多糖特异性ELISA中能够对1E4的结合产生超过70%的抑制。对恢复期人血清的研究表明,每毫升200微克的6F9能100%抑制C群特异性ELISA,每毫升50微克的6F9能实现该ELISA 50%的抑制。用Ab2(6F9)免疫BALB/c小鼠产生了特异性与Ab1、1E4相似的单克隆抗独特型抗体(Ab3)。用6F9免疫兔子产生了免疫球蛋白G反应,其效价显著高于对照组,达到1:160。这些研究表明,单克隆6F9在结合抗体位点上包含一个模拟脑膜炎球菌C多糖的替代影像。这个替代影像能够在同基因和异基因物种中引发针对脑膜炎球菌C多糖的抗体。