Generation of human monoclonal antibodies against colon cancer
K. Koda, M. C. Glassy and H. R. Chang
Department of Surgery, University of California San Diego Medical Center 92161.
Lymphocytes from regional lymph nodes of patients with colon cancer were
fused with a human lymphoblastoid cell line with or without in vitro
immunization. The efficacy of these two protocols for the generation of
human monoclonal antibodies against colon cancer was investigated. The
hyperplastic lymph nodes adjacent to the tumor were the best source of B
lymphocytes. Fusion frequency and the number of tumor-reactive clones were
markedly increased when the in vitro immunization protocol was applied
prior to fusion. As a stimulant in in vitro immunization, the supernatant
of pokeweed mitogen-stimulated T lymphocytes was superior to the
supernatant of mixed lymphocytes culture. Carcinoembryonic antigen at 20
micrograms/L seemed to be the optimal dose for in vitro immunization. The
reactivities of human monoclonal antibodies thus generated were measured by
enzyme-linked immunosorbent assay and confirmed by immunoperoxidase
staining. Combining in vitro immunization with lymphocytes of cancer
patients may lead to the successful production of clinically useful human
monoclonal antibodies.
