Chanh T C, Rappocciolo G, Hewetson J F
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, TX 78284.
J Immunol. 1990 Jun 15;144(12):4721-8.
An IgG1 mAb, designated HD11, specific for the trichothecene mycotoxin T-2 and capable of neutralizing its cytotoxicity was used to generate a syngeneic monoclonal anti-Id antibody. The generated anti-Id mAb, designated DE8, specifically bound to HD11 anti-T-2 mAb, and not to IgG1 mAb of irrelevant specificity or to normal mouse Ig. DE8 inhibited the binding of HD11 anti-T-2 to T-2-BSA-coated plates, whereas a control anti-Id mAb did not, suggesting recognition of an Id determinant associated with the T-2 binding site of HD11. Moreover, the binding of HD11 to DE8 and that of DE8 to HD11 were specifically inhibited by free T-2 mycotoxin. DE8 mAb was efficient in abrogating the protective effect of HD11 in the cytotoxicity of T-2 on the human epidermoid carcinoma cell line Hep-2. In vivo immunization of BALB/c mice with DE8 conjugated to KLH induced an anti-T-2 antibody titer comparable to that obtained with T-2-OVA immunization, whereas immunization with unconjugated DE8 resulted in a lower titered anti-T-2 response. Immunization with DE8-keyhole limpet or with unconjugated DE8 induced anti-T-2 antibody responses characterized by expression of "HD11-like" Id and by protection against T-2 cytotoxicity. However, the T-2-OVA-induced anti-T-2 response lacked the HD11+ Id and was only partially protective against T-2 cytotoxicity. This represents the first demonstration of the use of an anti-Id based vaccine in the in vivo induction of a protective antibody response against the cytotoxicity of a nonproteinaceous, small m.w. biologic toxin, whose very toxic nature precludes its use as the immunogen.
一种针对单端孢霉烯族霉菌毒素T-2且能够中和其细胞毒性的IgG1单克隆抗体（mAb），命名为HD11，被用于产生同基因单克隆抗独特型抗体。所产生的抗独特型mAb，命名为DE8，特异性结合HD11抗T-2 mAb，而不与无关特异性的IgG1 mAb或正常小鼠Ig结合。DE8抑制HD11抗T-2与包被有T-2-牛血清白蛋白（T-2-BSA）的平板的结合，而对照抗独特型mAb则无此作用，这表明识别了与HD11的T-2结合位点相关的独特型决定簇。此外，游离的T-2霉菌毒素可特异性抑制HD11与DE8的结合以及DE8与HD11的结合。DE8 mAb可有效消除HD11对T-2对人表皮样癌细胞系Hep-2细胞毒性的保护作用。用与钥孔戚血蓝蛋白（KLH）偶联的DE8对BALB/c小鼠进行体内免疫诱导出的抗T-2抗体效价与用T-2-卵清蛋白（T-2-OVA）免疫所获得的相当，而用未偶联的DE8免疫则导致抗T-2反应的效价较低。用DE8-匙孔血蓝蛋白或未偶联的DE8免疫诱导出的抗T-2抗体反应的特征是表达“HD11样”独特型并对T-2细胞毒性具有保护作用。然而，T-2-OVA诱导的抗T-2反应缺乏HD11 +独特型，并且仅对T-2细胞毒性具有部分保护作用。这首次证明了基于抗独特型的疫苗在体内诱导针对非蛋白质、低分子量生物毒素细胞毒性的保护性抗体反应中的应用，该毒素的剧毒性质使其无法用作免疫原。
