A human monoclonal antibody that detects HLA-A1, A23 and A24 antigens.
We report the production and characterization of a human monoclonal IgM (mu, kappa) antibody recognizing the HLA A1, A23 and A24 antigens. B lymphocytes obtained from a multiparous Japanese woman were transformed in vitro by Epstein-Barr virus, screened with an immune adherence assay, and fused with a murine myeloma cell line, P3-X63-Ag8.653. After subcloning by limiting dilution three times, a stable antibody-secreting hybridoma cell line, 4-35-7, was identified. The culture supernant had a titer of 1:32-64 against each of A1-, A23- and A24-positive lymphocyte panels, and showed complete correlation (r = 1.00) with the A1, A23 and A24 antigens on a lymphocyte panel of 287 unrelated, class I HLA-typed donors by the NIH cytotoxicity assay. Monoclonality of the antibody was ensured by Southern blot analysis of the human immunoglobulin heavy chain gene of 4-35-7. In view of the published data on HLA class I nucleotide sequences, the antibody may recognize an antigeneic determinant including two amino acid residues, Asp-166 and Gly-167, in the alpha 2 helix of the class I molecule that are specific for A1, A23 and A24 so far analyzed.
Kubo
K
K
Japanese Red Cross Hiroshima Blood Center.
Tachino
J
J
Yamaguchi
H
H
Kurachi
T
T
Naoe
T
T
Ohno
R
R
Ueda
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000166:cytology; D015139:Blotting, Southern; D002459:Cell Fusion; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D015234:HLA-A Antigens / Q000276:immunology* / Q000378:metabolism; D015788:HLA-A1 Antigen / Q000276:immunology* / Q000378:metabolism; D059871:HLA-A24 Antigen; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009101:Multiple Myeloma / Q000473:pathology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015234:HLA-A Antigens; D015788:HLA-A1 Antigen; D059871:HLA-A24 Antigen