CAJ | 학술논문

目的:制备鼠抗人vSIG4分子单克隆抗体(mAb)及对其生物学特性进行鉴定.方法:以高表达膜型VSIG4分子的基因转染细胞L929/VSIC4L为免疫原,常规免疫BALB/c小鼠,采用杂交瘤技术,经流式细胞术检测和多次;隆化培养,筛选出特异分泌鼠抗人VSIG4分子的杂交瘤细胞株;采用间接免疫荧光法、western blot、竞争抑制试验及MTT增殖试验等单对抗的生物学特征进行鉴定.结果:获得2株持续、稳定分泌鼠抗人VSIG4 mAb的杂交瘤细胞株,分别命名为9A7和9D5.对mAb生物学特性的研究结果表明,2株mAb均能识别巨噬细胞以及jurkat、THP-1、H446等肿瘤细胞株表面的VSIG4分子.对T细胞增殖抑制的阻断实验显示,这2株抗体对VSIG4分子抑制T细胞增殖均有阻断作用.结论:成功地获得了2株鼠抗人VSIG4功能性mAb,为进一步研究VSIG4及其未知受体的生物学功能奠定了基础.
목적:제비서항인vSIG4분자단극륭항체(mAb)급대기생물학특성진행감정.방법:이고표체막형VSIG4분자적기인전염세포L929/VSIC4L위면역원,상규면역BALB/c소서,채용잡교류기술,경류식세포술검측화다차;륭화배양,사선출특이분비서항인VSIG4분자적잡교류세포주;채용간접면역형광법、western blot、경쟁억제시험급MTT증식시험등단대항적생물학특정진행감정.결과:획득2주지속、은정분비서항인VSIG4 mAb적잡교류세포주,분별명명위9A7화9D5.대mAb생물학특성적연구결과표명,2주mAb균능식별거서세포이급jurkat、THP-1、H446등종류세포주표면적VSIG4분자.대T세포증식억제적조단실험현시,저2주항체대VSIG4분자억제T세포증식균유조단작용.결론:성공지획득료2주서항인VSIG4공능성mAb,위진일보연구VSIG4급기미지수체적생물학공능전정료기출.
Objective: To prepare anti-VSIG4 monoclonal antibodies and characterize their biological functions.Methods: BALB/c mice were immunized with transfected cell line (L929/VSIG4L) as immunogen.The spleen B cells of the mice were fused with SP2/0 and hybridoma cells were screened with transfected cell line (L929/VSIG4) by FCM.After acquisition of the hybridomas secreting anti-VSIG4 mAb,their biological activities were investigated by indirect immunofluorescence,Western blot,competitive inhibition test,and MTT assay.Results:Two stable hybridomas,9A7 and 9D5 were obtained,which could continuously secrete specific anti-VSIG4 monoclonal antibodies.The following biological activity studies showed that these monoclonal antibodies could recognize the natural VSIG4 expressed on the macrophages and several cancer cell lines,such as Jurkat,THP-1 and H446.Furthermore,they could block the inhibitory effects of VSIG4 on proliferation of T cells in vitro.Conclusion: Two hybridomas secreting anti-VSIG4 monoclonal antibodies have been established.These monoclonal antibodies provide useful tools for further studying VSIG4's biological function and its unknown receptor.