title: Rearrangement of the human heavy chain variable region gene V3-23 in transgenic mice generates antibodies reactive with a range of antigens on the basis of VHCDR3 and residues intrinsic to the heavy chain variable region.
abstract: To formulate a 'logic' for how a single immunoglobulin variable region gene generates antibodies with different antigen specificity and polyreactivity, we analysed chimeric antibodies produced in transgenic mice carrying the germ-line human V3-23 gene, multiple diversity (D) and joining (J) gene segments. Hybridomas producing antibodies encoded by the V3-23 gene in combination with different mouse Vkappa genes were obtained by fusion of splenocytes from transgenic mice. All antibodies had human mu-chains and mouse light chains, were multimeric in structure and expressed the human V3-23 gene. Nucleotide sequence analyses of genes encoding the heavy and light chains of 12 antibodies in relation to antigen specificity highlighted the importance of heavy chain variable region CDR3 in determining reactivity with different antigens. However, the results also suggest that non-CDR3 sequences intrinsic to the V3-23 gene itself may be involved in, or determine, the binding of the chimeric antibodies to some of the antigens tested in the current study.
authors.affiliation: Kennedy Institute of Rheumatology and Rheumatology Unit, Charing Cross Hospital, London, UK. r.mageed@cxwms.ac.uk
authors.forename: R A
authors.initials: RA
authors.lastname: Mageed
authors.forename: I J
authors.initials: IJ
authors.lastname: Harmer
authors.forename: S L
authors.initials: SL
authors.lastname: Wynn
authors.forename: S P
authors.initials: SP
authors.lastname: Moyes
authors.forename: B B
authors.initials: BB
authors.lastname: Maziak
authors.forename: M
authors.initials: M
authors.lastname: Brüggemann
authors.forename: C G
authors.initials: CG
authors.lastname: MacKworth-Young
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis* / Q000378:metabolism; D000937:Antigen-Antibody Reactions / Q000235:genetics*; D001483:Base Sequence; D002459:Cell Fusion / Q000379:methods; D022801:Complementarity Determining Regions / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain / Q000276:immunology*; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain / Q000276:immunology; D005803:Genes, Immunoglobulin / Q000276:immunology*; D018095:Germ-Line Mutation; D006801:Humans; D006825:Hybridomas; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid
chemical_list: D000888:Antibodies, Anti-Idiotypic; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains