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Keiji Koda, MD; Mark C. Glassy, PhD; Helena R. Chang, MD, PhD

Arch Surg. 1990;125(12):1591-1597.

Abstract
• 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 µg/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.

(Arch Surg. 1990;125:1591-1597)

Author Affiliations
From the Departments of Surgery, University of California San Diego Medical Center and the Veterans Affairs Medical Center (Drs Koda and Chang), and BioTechnetics (Dr Glassy), San Diego.

Footnotes
Accepted for publication August 30, 1990.

Read before the 14th Annual Surgical Symposium of the Association of Veterans Affairs Surgeons, Charleston, SC, May 8, 1990.

Reprint requests to UCSD Medical Center and VAMC, V-112H, 3350 La Jolla Village Dr, San Diego, CA 92161 (Dr Chang).

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