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在前阶段工作中已获得16个抗天花粉蛋白的单抗。用其中的一个IgE类单抗TE 1免疫Wistar大鼠,通过大鼠-小鼠杂交瘤技术得到了抗独特型单抗。现研究抗独特型单抗AId 6c5在体外诱发二次抗体应答中所起的作用。实验结果表明:1.当将AId 6c5和天花粉蛋白初次免疫8周后的小鼠脾细胞和肠系膜淋巴结细胞一起培养时,能引起二次抗体应答,说明AId 6c5能代替抗原的刺激作用,如果培养系统中同时存在AId 6c5和天花粉蛋白(其剂量能引起体外二次抗体应答),则可出现某种程度的抑制。由于AId 6c5是单克隆抗体,提示一种AId能在不同情况下起刺激或抑制作用。3.应用竞争结合试验阐明AId 6c5除和TE 1外,还和另外两个单抗——TE 4(IgE)和2 A1(IgG1)——起反应,先前的工作证明这三个单抗和另外4个单抗都识别天花粉蛋白上的同一抗原决定簇。但AId 6c5对识别这同一决定簇的其余4个单抗反应很弱。以上说明a.IgE和其它Ig类别具有共同或交叉的独特型,b.也说明识别同一决定簇的IgE具有不同的独特型。4.将TE 1等三个单抗和AId 6c5预先作用后能抑制这三个单抗和天花粉蛋白的结合,说明AId 6c5所识别的独特型,位于抗原结合部位内,至少??是很靠近抗原结合部位的。
In the previous phase of work, 16 anti-Trichosanthin monoclonal antibodies have been obtained. Wistar rats were immunized with one of the IgE monoclonal antibodies TE 1 and anti-idiotypic monoclonal antibodies were obtained by the rat-mouse hybridoma technique. We now study the role of anti-idiotypic AIb 6c5 in inducing secondary antibody responses in vitro. The results showed as follows: 1. When the spleen cells and mesenteric lymph node cells of mice immunized with AId 6c5 and trichosanthin for 8 weeks were elicited secondary antibody response, indicating that AId 6c5 can replace the stimulation of antigen, There is some degree of inhibition of AId 6c5 and trichosanthin present in the system at doses that cause secondary antibody responses in vitro. Since AId 6c5 is a monoclonal antibody, it suggests that an AId can stimulate or inhibit in different situations. 3. Using competition-binding assays to elucidate that AId 6c5 reacts with two other mAbs, TE 4 (IgE) and 2 A1 (IgG1), in addition to TE 1 and previous work demonstrated that these three mAbs and The other four monoclonal antibodies recognize the same epitope on trichosanthin. However, AId 6c5 reacts weakly to the other 4 mAbs that recognize this same determinant. The above demonstrates that a. IgE and other Ig classes have a common or overlapping idiotype and b. Also indicates that IgE recognizing the same determinant has a distinct idiotype. 4. The pretreatment of three monoclonal antibodies TE 1 and AId 6c5 can inhibit the combination of these three monoclonal antibodies and trichosanthin, indicating that the idiotype recognized by AId 6c5 is located in the antigen-binding site, at least very close Antigen binding site.