Affiliations: [a] Guildhay Antisera Ltd., Unit 6, Riverside Business Centre, Walnut Tree Close, Guildford, Surrey GUI 4UG, UK | [b] Department of Immunology, St. Thomas' Hospital, London, SE1 7EH, UK
Note: [] Address reprint requests to Dr. Mark G. De Silva, Guildhay Antisera Ltd., Unit 6, Riverside Business Centre, Walnut Tree Close, Guildford GU1 4UG, Surrey, UK.
Abstract: Human monoclonal antibodies have been produced from lymphocytes of an acute-onset insulin-dependent diabetic patient. Peripheral blood lymphocytes were hybridized with a fusion partner HMY-1320. Initial screening of human immunoglobulin secretion was made by a nitrocellulose dot blot assay. Ten stable cell lines of novel type, secreting human immunoglobulin, were obtained. These cell lines have been maintained in continuous culture over 6 months and cryopreserved in liquid nitrogen for 14 months. Human monoclonal antibodies of IgG and IgM class have been produced and are secreted at a rate of 150–650 ng/ml/106 cells/day. Monoclonal antibodies were tested for histological staining against a variety of endocrine and non-endocrine tissues. One monoclonal antibody, LT1E12, demonstrates a staining pattern in human, rat, and mouse tissues, similar to that of mitochondrial antibodies. Another antibody, LT3C4, demonstrates weak staining of smooth muscle in rat and mouse kidney sections. Neither specificities were detected in the diabetic patient's serum. The variety of immune tissue specificities obtained in this study demonstrates the potential value of human monoclonal antibodies as probes to analyze the complexity of autoimmunity in diabetes mellitus.
Keywords: human monoclonal antibodies, novel cell lines, diabetes mellitus, autoimmunity
