Fernández I M, Ovaa W, Harmsen M, Benaissa-Trouw B J, Bos N A, Kraaijeveld C A, Snippe H
Eijkman-Winkler Institute of Medical and Clinical Microbiology, University Hospital Utrecht, The Netherlands.
Viral Immunol. 1994;7(2):71-80. doi: 10.1089/vim.1994.7.71.
In the present study a shared idiotope was found among antibodies against a previously defined linear B-cell epitope of Semliki Forest virus (SFV). The synthetic B-cell epitope, located at amino acid positions 240 to 255 of the E2 membrane protein, was linked to an H-2d-restricted T-helper cell epitope of either SFV or influenza virus. Colinearly synthesized peptides of T-B polarity mixed with adjuvant were used to immunize BALB/c (H-2d) mice. After one booster immunization with either chimaeric peptide high serum antibody titers were measured against both synthetic peptide (240-255) and glutaraldehyde-fixed SFV-infected L cells. Against the synthetic peptide (240-255) a variety of monoclonal antibodies (MAbs) were produced that differed in reactivity with SFV, varied in heavy chain family, isotype, isoelectric point, and idiotype. Against one of the antipeptide MAbs (I02), that strongly reacted with SFV-infected L cells, an antiidiotypic MAb (ab2MAb), designated I02A3, was produced that could be inhibited in its binding to MAb I02 by the synthetic B-cell epitope. Therefore it was concluded that ab2 MAb I02A3 recognizes an idiotope closely associated with the antigen combining site of antipeptide MAb I02. This idiotope was definitively shared by two out of 15 antipeptide MAbs and by SFV-reactive antibodies present in both antipeptide sera and SFV-immune sera.
在本研究中,发现针对先前确定的Semliki森林病毒(SFV)线性B细胞表位的抗体之间存在一个共同的独特型。位于E2膜蛋白氨基酸位置240至255的合成B细胞表位与SFV或流感病毒的H-2d限制性T辅助细胞表位相连。将具有T-B极性的共线性合成肽与佐剂混合用于免疫BALB/c(H-2d)小鼠。用嵌合肽进行一次加强免疫后,检测到针对合成肽(240-255)和戊二醛固定的SFV感染的L细胞的高血清抗体滴度。针对合成肽(240-255)产生了多种单克隆抗体(MAb),它们与SFV的反应性不同,重链家族、同种型、等电点和独特型也各不相同。针对其中一种与SFV感染的L细胞强烈反应的抗肽单克隆抗体(I02),产生了一种抗独特型单克隆抗体(ab2MAb),命名为I02A3,其与单克隆抗体I02的结合可被合成B细胞表位抑制。因此得出结论,ab2单克隆抗体I02A3识别与抗肽单克隆抗体I02的抗原结合位点密切相关的独特型。15种抗肽单克隆抗体中有2种以及抗肽血清和SFV免疫血清中存在的SFV反应性抗体明确共享了这种独特型。