抗旋毛形线虫单克隆抗体的制备和抗原定位研究  

Preparation of Monoclonal Antibodies against Trichinella spiralis and Localization of Antigens

作　　者：冯瑞元[1] 阎和平[1] 刘英[2] 

机构地区：[1]华西医大寄生虫学教研室 [2]成都军区陆军总医院

出　　处：《华西医科大学学报》1990年第1期1-4,共4页Journal of West China University of Medical Sciences

基　　金：国家自然科学基金~~

摘　　要：作者用旋毛形线虫感染期幼虫免疫的BALB/c小鼠脾细胞,与SP2/0骨髓癌细胞进行杂交。融合率92.1％,抗体阳性率16.6％,通过克隆化已初步建立分泌抗幼虫可溶性抗原的单克隆抗体(McAb)的杂交瘤细胞18株。采用间接过氧化物酶法,以证实能特异地识别此虫感染期幼虫抗原组分的3株McAb定位抗原,揭示相关抗原组分主要位于幼虫的角度层、杆状体,其次为肠道。BALB/c mice were infected per os withthe infective larvae(L1) of Trichinella spi-ralis, and chellenged by injecting 0.2mlsoluble L1 antigen 3 days before cell fusion.SP2/0 myeloma and immune spleen cellswere fused at the ratio of 1:5 in the pre-sence 30%PEG (MW 4000). The fusioll rateand antibody positive rate were 92.1%and 16.6% respectively using ELISA.Amongthese the chromosomes of 4 strains were2n>100 and that of the SP2/o myelomacells 2n≤70. The titer of McAb in the asciteswas found to be 1/640 or 1/1 280. Eight strainswere of IgG1, 1 IgG2a and 3 IgM in an agarsystem. Three strains of McAbs, Which couldrecognize specifically the L1 antigen frac-tions by immuno-blotting technique, wereused as probes to localize the target anti-gens in sections of T. spiralis L1. Stainingwas performed using an indirect techniqueconsisting of goat anti-mouse IgG-conjuga-ted horseradish peroxidase on de-paraffinedsections, and substrate DAB. The resultsrevealed that the antigens reacted moststrongly with the specific McAbs werelocated in the cuticle and stichosome,followed by the lining of gut. Among the 3strains of McAbs used, 2E_5F_(11)A_5 reactedmost strongly with the antigen, followedby 2E_5E_9E_4. 2E_5E_9G_5 was the weakest.

关 键 词：旋毛形线虫 单克隆抗体 抗原定位 

分 类 号：R392－33[医药卫生—免疫学]