Effective induction and acquisition of human monoclonal IgE antibodies reactive with house-dust mite extracts

IgE plays a critical role in acute hypersensitivity such as anaphylaxis, as thma, and atopic dermatitis. IgE antibody is, therefore, an essential reage nt for studying the mechanisms of these diseases. However, it is difficult to obtain IgE antibody in amounts sufficient for research use because IgE-p roducing lymphocytes are very rare. To overcome this problem, we investigat ed the requirements for generating IgE-secreting human hybridomas using in vitro immunization of peripheral blood lymphocytes. First, culture conditio ns were optimized for IgE production by a combination of the immunomodulato ry mediators interleukin-2, interleukin-4, interleukin-6, and muramyl dipep tide. Second, the addition of mite antigen to the cultures resulted in an i ncreased production of antigen-specific IgE as well as antigen-specific IgG and IgM. When activated lymphocytes in these cultures were fused with Burk itt lymphoma cells, ICLU-B, antigen-specific IgE-secreting hybridomas were obtained with high efficiency. These results demonstrate that our culture a nd in vitro immunization system for human peripheral blood lymphocytes is u seful for obtaining antigen-specific IgE. (C) 2000 Elsevier Science B.V. Al l rights reserved.