Immunologic and functional characterization of anti-HLA-DR rabbit antibodies induced by synthetic peptides |
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Authors: | A Chersi G Schulz R A Houghten |
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Affiliation: | 1st. Istituto Regina Elena for Cancer Research, Viale Regina Elena 291, 00161 Roma, Italy;1. The Children''s Hospital, University of Giessen, F.R.G.;2. Scripps Clinic and Research Foundation, La Jolla, CA, U.S.A. |
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Abstract: | Three peptides selected from the amino acid sequence of the alpha- and beta-chains of DR2 histocompatibility antigens were chemically synthesized and coupled to carrier proteins to be used as immunogens in rabbits. This immunization resulted in the production of specific antibodies that readily recognized the antigen. However, only one of the four antibody preparations, antibody 6148, elicited by a short peptide from the beta-chain (residues 61-73), reacts with native membrane glycoproteins as well as intact human lymphoblastoid cells in enzyme-linked immunosorbant assays. This antibody was found to react also with membrane glycoproteins solubilized by nonionic detergents from cells bearing a different HLA-DR specificity: therefore it is likely that the peptide responsible for eliciting antibody 6148 represents a common framework determinant of DR alloantigens that is accessible on the surface of lymphoblastoid cells. The ability of antibody 6148 to bind to intact cells was confirmed by indirect immunofluorescence and by fluorescein-activated cell sorter analysis. This antibody is also capable of mediating antibody-dependent cellular cytotoxicity as determined by a 51Cr-release assay. |
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Keywords: | KLH keyhole limpet hemocyanin MBS PBS phosphate-buffered saline SDS-PAGE sodium dodecyl sulfate-polyacryiamide gel electrophoresis TMD tetramethylendiamine FCS fetal calf serum ADCC antibody-dependent cellular cytotoxicity FACS fluorescein-activated cell sorter |
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