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  • Title: Production of human monoclonal antibodies to B lymphocyte cell surface antigens by in vitro immunization and human-human hybridoma technology.
    Author: Power DA, Gerkis V, d'Apice AJ.
    Journal: Hum Antibodies Hybridomas; 1990; 1(1):34-41. PubMed ID: 2103351.
    Abstract:
    We attempted to produce human monoclonal IgM antibodies to class II HLA antigens by in vitro immunization of human splenocytes with affinity purified HLA or irradiated whole lymphoblastoid B cells using the adjuvant muramyl dipeptide and IL-2. Two fusions where affinity purified HLA was used to stimulate the in vitro immunization produced no hybrids secreting antibodies against the stimulating antigen. However, 13.1% of 381 hybrids from eight fusions were positive by CELISA when whole cells were used to immunize. These antibodies were reactive with a wide range of cell types and were not directed to HLA. It proved difficult to characterize them, possibly because of low affinity. One reacted in Western blots with a 200 kD antigen, but this was exceptional. Most bound to a small percentage of cells in flow cytometry and did not lyse target cells in cytotoxicity assays. Control fusions were performed and similar antibodies were obtained from 2.6% of 235 hybrids. These data suggest that in vitro immunization using the protocol outlined can increase the frequency of some antibodies, but that they may represent a very primitive lineage which does not have the specificity and affinity required to prove useful as diagnostic or therapeutic agents.
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