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天花粉蛋白是中药天花粉的有效引产成份,在应用中它偶尔也引起过敏反应。我们用杂交瘤技术建立了一株分泌抗天花粉蛋白特异性IgE单克隆抗体的杂交瘤细胞系。在实验中,二次免疫的C57 BL/6 J小鼠的肠系膜淋巴结细胞和脾细胞分别被用来同NSI骨髓瘤细胞进行融合。虽然在融合前动物血清IgE抗体效价仅为40～160 PCA滴度,但用肠系膜淋巴结细胞进行的4次融合都产生了IgE杂交瘤。阳性率为1.0-6.7%。对比之下,用脾细胞进行的另外两次融合却没有观察到IgE杂交瘤产生,统计结果说明差异显著(P<0.05)。该IgE单克隆抗体可以在体内和体外诱发大鼠肥大细胞脱颗粒。56℃热处理2小时能使该抗体诱导PCA反应的能力完全丧失,然而却不影响其结合抗原的能力。对该单克隆抗体的特异性的鉴定表明,它可以特异性地识别精制天花粉蛋白和结晶天花粉蛋白上的抗原决定簇。
Trichosanthin is an effective induction of TCS ingredients, which occasionally causes allergic reactions. We have established a hybridoma cell line that secretes anti-Trichosanthin specific IgE monoclonal antibodies using hybridoma technology. In experiments, mesenteric lymph node cells and splenocytes from double immunized C57BL / 6J mice were used to fuse with NSI myeloma cells, respectively. Although the serum IgE antibody titer was only 40-160 PCA titers prior to fusion, all four confluence with mesenteric lymph node cells yielded IgE hybridomas. The positive rate of 1.0-6.7%. In contrast, no IgE hybridomas were observed for the other two rounds of fusion with splenocytes, with statistical significance indicating significant differences (P <0.05). The IgE monoclonal antibody can induce degranulation of rat mast cells in vivo and in vitro. Heat treatment at 56 ° C for 2 hours completely abolished the ability of the antibody to induce a PCA reaction without affecting its ability to bind antigen. The specific identification of this monoclonal antibody shows that it can specifically recognize antigenic determinants on both refined trichosanthin and crystalline trichosanthin.