Production and characterization of two murine monoclonal antibodies directed against epitopes exclusive to soluble fragments of Fc epsilon RII/CD23.
Two murine monoclonal antibodies (mAb) designated as SU1 and SU3 directed against soluble Fc epsilon RII/CD23 have been generated by fusing X.63.AG.8653 (a mouse myeloma cell line) with spleen cells from mice immunized with an Epstein Barr virus (EBV)-transformed B cell line (RPMI-8866). The antibodies have been shown to be capable of detecting affinity purified soluble Fc epsilon RII/CD23 in an enzyme-linked immunosorbent assay. Indirect immunofluorescence has shown that the SU1 and SU3 mAb do not stain RPMI-8866, a Fc epsilon RII/CD23+ B cell line. By studying the migration profiles of affinity purified SU1- and SU3-reactive molecules on sodium dodecylsulfate-polyacrylamide gel electrophoresis it has been shown that SU1 mAb immunoprecipitates 33- and 12-kDa components, while the SU3 mAb recognized 25- and 45-kDa proteins from culture supernatants of RPMI-8866 cells. Moreover, affinity purified SU1- and SU3-reactive proteins have been shown to be recognized by human IgE but not by the human IgG molecule. These results provide evidence that SU1 and SU3 mAb may recognize some putative post-cleavage epitopes on the N-terminal end of the low affinity receptor which appear, perhaps, following the process of fragmentation. In addition, the effect of these antibodies on continuous growth of a panel of lymphoblastoid cell lines indicates that SU1 mAb was found incapable of influencing the spontaneous proliferation of EBV-immortalized B cell lines; whereas SU3 mAb completely blocked the spontaneous growth and proliferation of all B cell lines tested. The results are discussed in relation to the appearance of a functional post-cleavage epitope on soluble Fc epsilon RII/CD23.
Ghaderi
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AA
Department of Microbiology and Immunology, Medical School, Shiraz University.
Stanworth
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DR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001665:Binding Sites; D002455:Cell Division; D002478:Cells, Cultured; D000939:Epitopes / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007073:Immunoglobulin E / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000276:immunology / Q000378:metabolism; D017455:Receptors, IgE / Q000276:immunology* / Q000378:metabolism; D012995:Solubility
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D017455:Receptors, IgE; D007073:Immunoglobulin E