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目的制备抗泡球蚴组织单克隆抗体,并鉴定其特异性。方法运用泡球蚴组织粗抗原免疫小鼠制备免疫脾细胞,运用淋巴细胞杂交瘤技术将其与小鼠骨髓细胞SP2/0融合。采用ELISA和免疫组化方法筛选抗泡球蚴原头节单克隆抗体的杂交瘤细胞株。检测所获细胞株的染色体数目,采用ELISA和免疫组化法鉴定所获抗体的特异性。结果获得了一株能稳定分泌的抗泡球蚴原头节单克隆抗体的杂交瘤细胞株6G2A7F7。染色体数目为98条,为明显的融合细胞核型。所分泌的抗体McAbP325能与泡球蚴中的生发层和原头节特异结合,特别对原头节上的小钩和吸盘表现出很强的结合反应。但与棘球蚴(人源)和水泡带绦虫幼虫(细颈囊尾蚴)组织无特异性结合反应。结论制备的杂交瘤细胞能稳定分泌McAbP325,为泡球蚴的细胞分类、免疫组化和抗原研究提供工具和基础。
Objective To prepare monoclonal antibodies against Cysticercus cellulosae and to identify its specificity. Methods The immunized spleen cells were prepared by immunizing mice with crude antigen of Echinococcus granulosus and fused with mouse bone marrow cells SP2 / 0 using lymphocyte hybridoma technique. ELISA and immunohistochemistry were used to screen the hybridoma cell line of the monoclonal antibody against the prothallus of Echinococcus multilocularis. The number of chromosomes of the obtained cell lines was detected, and the specificity of the obtained antibodies was identified by ELISA and immunohistochemistry. As a result, a hybridoma cell line 6G2A7F7 capable of stably secreting the precursor monoclonal antibody against Cysticercus cellulosae was obtained. The number of chromosomes is 98, which is obvious fusion karyotype. The secreted antibody McAbP325 can combine with the germinal layer and the protuberant section of the hydatid cyst, especially showing the strong binding reaction between the small hook and the sucker on the head. But no specific binding reaction with hydatid cyst (human source) and Talarlus taenia larvae (Cysticercus cellulosae) tissues. Conclusion The prepared hybridoma cells can stably secrete McAb325 and provide the tools and basis for the cell classification, immunohistochemistry and antigenic study of Cysticercus cellulosae.