新型免疫佐剂在淋巴细胞杂交瘤技术中的应用  

Application of a new adjuvant (CpG ODN)in the preparation of lymphocyte hybridoma technique

作　　者：许晓光[1] 朱勇[1] 刘雪松[1] 刘莹[1] 户义[1] 金伯泉[1] 

机构地区：[1]第四军医大学免疫学教研室,陕西西安710032

出　　处：《免疫学杂志》2003年第6期419-423,共5页Immunological Journal

基　　金：教育部"高等学校骨干教师资助计划"项目 (2 0 0 0年度 )

摘　　要：目的　观察新型免疫佐剂ImmunEasyTM(CpGODN)的免疫调节作用 ,并与弗氏佐剂相比较 ,探讨其在单克隆抗体制备中的应用前景。方法　将小鼠用新型免疫佐剂ImmunEasyTM 或弗氏佐剂混合原核表达的重组APRIL抗原 ,分别以常规免疫和少量抗原免疫方案免疫Balb c小鼠 ,用间接ELISA法检测不同时相小鼠免疫血清抗APRIL抗体的总效价及各类和亚类的效价。建立一种能在单个细胞水平反映小鼠B细胞分泌特异性抗体亚类的改良ELISPOT方法 ,并对两种佐剂免疫组中配对的小鼠进行检测。常规方法制备杂交瘤 ,以间接ELISA法筛选阳性杂交瘤 ,比较不同佐剂免疫小鼠所获得的杂交瘤上清液的类及其亚类频率。结果　与弗氏佐剂相比 ,ImmunEasyTM 在单克隆抗体制备过程中可明显减少抗原用量 ,缩短免疫周期 ,提高免疫血清效价 ,并增加IgG2a、IgM及IgA抗体的比例。结论　在细胞融合制备鼠源性mAb中 ,应用ImmunEasyTM,可减少免疫原用量 ,提高效率 ,还可优先获得某些类及其亚类的mAb。Objective To observe the effect of immunological regulation and the advantage of the mouse adjuvant ImmunEasy compared with Freund's adjuvant in the preparation of lymphocyte hybridoma, and investigate its potential application in the preparation of monoclonal antibody. Methods Balb/c mice were immunized with serial doses of APRIL protein mixed with ImmunEasy or Freund's adjuvant, respectively. The titers and isotypes of the specific antibodies in the mouse sera from different groups were compared periodically by indirect ELISA and modified ELISPOT. The splenocytes from immunized mice were fused with SP2/0 cells and the positive hybridoma clones were screened by indirect ELISA. And the supernatant from the hybridoma clones were used to detect the isotypes of immunoglobulins. Results Compared with Freund's adjuvant, ImmunEasy can remarkably save immunogen, shorten immunization period, increase titers of total specific antibody and that of IgG2a, IgM and IgA in the sera of mice. Conclusion In the preparation of mAbs, ImmunEasy can save immunogen, improve the efficiency, and get mAbs of some isotypes preferentially.

关 键 词：免疫佐剂 淋巴细胞杂交瘤 免疫调节作用 ImmunEasy^TM 单克隆抗体 

分 类 号：R392[医药卫生—免疫学] R730.26[医药卫生—基础医学]