抗sAPRIL突变体多抗的制备及免疫抑制功能分析  

Preparation and immunity suppression function of polyclonal antibody against sAPRIL mutant antigen

作　　者：李强[1] 高全胜[2] 刘莹[1] 

机构地区：[1]第三军医大学新桥医院检验科,重庆400037 [2]第三军医大学基础医学院病原生物学教研室,重庆400038

出　　处：《重庆医学》2006年第18期1633-1634,1637,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(30300314;30500622)

摘　　要：目的制备抗sAPRIL突变体多抗。观察其对sAPRIL促细胞增殖活性的影响。方法采用在sAPRILC端缺失45bp并加入HEL Th表位序列．且已经证实丧失sAPRIL天然活性的重组突变体蛋白，加上佐剂免疫BALB／c小鼠．6周后取血清。间接ELISA测定抗体滴度；Western blot测定抗体特异性；MTT检测所获抗血清抑制sAPRIL促Raji和Jurkat细胞增殖的作用。结果sAPRIL突变体能诱导小鼠产生特异性抗体，抗体具有较好的特异性，滴度达到1：10000～1：100000；抗sAPRIL突变体血清可明显降低sARPIL刺激Raji和Jurkat细胞增殖的吸光度值（P〈0．01）。结论成功获得抗sAPRIL突变体的多克隆抗体，该抗体可以在体外抑制sAPRIL促Raji和Jurkat细胞的增殖活性。Objective To prepare the polyclonal antibody against the mutant of soluble a proliferation-inducing ligand（sAPRIL） antigen and to clarify the inhibition of polyclonal antibodies on cell proliferation stimulated by sAPRIL. Methods Mice were immunized by purified proteins,mutant of sAPRIL modified by HEL Th epitopes with deletions at the C-terminals of 45bp were confirmed to lose the functional activity of normal sAPRIL and immunoadjuvant. Serum of mice were obtained after 6 weeks. Indirect ELISA and Western blot analyses confirmed the antibody titer and specificity. The inhibition of polyclonal antibodies on Raji and Jurkat cells proliferation stimulated by sAPRIL was assessed by the MTT assay. Results The mutant of sAPRIL could induce production of polyclonal antibodies against the sAPRIL mutant antigen. Western blot and indirect ELISA analysis showed that the anti-serum had higher specificity,and its titer were 1：10 000-1：100 000. Absorbance were markely decreased（P〈0.01 ） by polyclonal antibodies in MTT test of Raji and Jurkat cell proliferation stimulated by sAPRIL. Conclusion Polyclonal antibody against sAPRII, mutant antigen is obtained successfully, and the antiserum against mutant protein of sAPRII, may inhibit Raji and Jurkat cell proliferation stimulated by sAPRIL in vitro.

关 键 词：可溶性增殖诱导配体 突变体 抗体 免疫抑制 

分 类 号：R392[医药卫生—免疫学]