Characterization of three human monoclonal antibodies specific for polymorphic class i and class Ii HLA antigens

Dessi, V.; Sanchez, B.; Garzon, M.; Magariño, R.; Maldonado, M.D.; Nuñez-Roldan, A.

Human Immunology 27(4): 323-332


ISSN/ISBN: 0198-8859
PMID: 2156787
DOI: 10.1016/0198-8859(90)90083-2
Accession: 039526718

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Three human monoclonal antibodies were derived from a single polytransfused patient awaiting renal transplantation. In microcitotoxicity assays, the patient's serum displayed strong positive reactions against greater than 90% of a panel of cells representing the known HLA specificities. The donor's peripheral blood lymphocytes were infected with Epstein-Barr virus, cloned, and supernatants of the virus transformed cultures were screened for the presence of IgG antilymphocyte reactivity utilizing an enzyme-linked immunosorbent assay method. Positive cultures were recloned and fused with the human-mouse heteromyeloma SHM. Supernatants from three clones were selected for alloreactivity and characterized by indirect immunofluorescent staining and fluoroactivated cell sorter analysis on homozygous typing cells, including those from the Tenth International Histocompatibility Workshop core panel and on cell lines derived from selected families. Data obtained demonstrate that two human monoclonal antibodies have DQw1 specificity, one of them being reactive against several DQw7-positive cell lines, while one monoclonal antibody is specific for the A2 + A28 class I MHC antigens. Anti-DQw1 antibodies were of different light-chain subtypes.