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Production and characterization of human monoclonal antibodies against Schistosoma mansoni
Authors:ALFREDO M. GOES &Dagger  ,ROBERTO S. ROCHA&dagger  ,GIOVANNI GAZZINELLI&dagger  ,BARBARA L. DOUGHTY
Affiliation:Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Texas A & M University, College Station 77843.
Abstract:
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.
Keywords:Schistosoma mansoni,    antigen preparations,    human monoclonal antibodies,    mouse–human heteromyeloma,    Epstein–Barr virus cell lines
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