Goes A M, Rocha R S, Gazzinelli G, Doughty B L
Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Texas A & M University, College Station 77843.
Parasite Immunol. 1989 Nov;11(6):695-711. doi: 10.1111/j.1365-3024.1989.tb00930.x.
We have produced a panel of human monoclonal antibodies (MoAb) from patients infected with Schistosoma mansoni in order to analyse more carefully the human immune response to this helminth infection. This study describes the production, characterization and analysis of these MoAbs. Briefly, peripheral blood mononuclear cells from chronically infected patients were (1) isolated and stimulated with parasite antigens in vitro, (2) positively selected for B-cells on anti-Ig columns, and (3) then transformed with Epstein-Barr virus (EBV). Once EBV cell lines were established, they were selected for anti-S. mansoni antibodies using an ELISA, cloned, retested and then fused with the mouse-human heteromyeloma SHM-D33. In this study, we describe five MoAbs which have different antigenic specificities for life-cycle stages based on ELISA to soluble crude antigen preparations, membrane immunofluorescence on whole intact organisms, and immunofluorescent staining of cryostat frozen sections. The importance of these reagents with regard to the human immune response to S. mansoni is currently being evaluated.
为了更仔细地分析人体对曼氏血吸虫感染的免疫反应，我们从感染曼氏血吸虫的患者中制备了一组人单克隆抗体（MoAb）。本研究描述了这些单克隆抗体的制备、特性鉴定及分析。简要地说，从慢性感染患者的外周血单个核细胞中：（1）分离并在体外使用寄生虫抗原进行刺激；（2）在抗Ig柱上对B细胞进行阳性选择；（3）然后用爱泼斯坦-巴尔病毒（EBV）进行转化。一旦建立了EBV细胞系，便使用酶联免疫吸附测定（ELISA）筛选抗曼氏血吸虫抗体，进行克隆、重新检测，然后与小鼠-人异源骨髓瘤SHM-D33进行融合。在本研究中，我们描述了五种单克隆抗体，基于对可溶性粗抗原制剂的ELISA、对完整生物体的膜免疫荧光以及对低温恒温器冷冻切片的免疫荧光染色，它们对生命周期各阶段具有不同的抗原特异性。目前正在评估这些试剂对于人体对曼氏血吸虫免疫反应的重要性。
