In Vitro Immunization of Human Peripheral Blood Lymphocytes with Cholera Toxin B Subunit

Abstract

In vitro immunization (IVI) techniques have a great potential in the production of human monoclonal antibodies (MAbs) against various antigens. An IVI method of human peripheral blood lymphocytes (PBL) has been developed with a human lung adenocarcinoma cell line in our laboratory. We have succeeded in generating several cancer specific human MAbs using this IVI method. Because this IVI method was not available for soluble antigens, we improved it for soluble antigens. IVI with soluble antigens was effectively caused by the addition of muramyl dipeptides, interleukin-2 (IL-2) and interleukin-4 (IL-4). It wasfound thatthe difference of sensitivityof lymphocytes depending upon donors could be overcome to find the optimal concentrations of IL-2 and IL-4. IVI of human PBL was performed with cholera toxin B subunit (CTB) and the immunized B cells were transformed by Epstein-Barr virus. Anti-CTB antibody was detected using an indirect ELISA. Clones producing anti-CTB antibodies were directly cloned by a soft agar cloning method.KeywordsPeripheral Blood LymphocyteCholera ToxinHuman Monoclonal AntibodySoluble AntigenHuman Peripheral Blood LymphocyteThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.