The use of lymphocytes from axillary lymph nodes of mastectomy patients to generate human monoclonal antibodies

Abstract

Lymphocytes from axillary lymph nodes of breast cancer mastectomy patients were fused with murine non-immunoglobulin-secreting myeloma cells to generate hybridoma cell lines that synthesize human immunoglobulins. Lymph node lymphocytes from 21 patients were used to obtain 505 human-mouse hybrid cultures. From these, 62 cultures were established which synthesized immunoglobulins reactive in radioimmunoassays specific for either human IgG or human IgM. Some of these double-cloned hybrid cell lines produced human monoclonal antibodies for at least 6 months. Sodium dodecylsulfate polyacrylamide gel electrophoresis and immunodiffusion analysis demonstrated that the monoclonal antibodies possessed human heavy and light immunoglobulin chains. Levels of synthesis ranged from 0.1 to 20 μg of human immunoglobulin per ml of culture fluid. The immunoreactivity of some of these human monoclonal antibodies with mammary carcinoma cells is summarized and has been documented elsewhere ( J. Schlom, D. Wunderlich and Y. A. Teramoto . Proc. Natl Acad Sci USA 1980; 77: 6841 ); the reactivity of the majority of the immunoglobulins, however, is still unknown at this time. The studies reported here detail the procedures in which axillary lymph nodes from mastectomy patients are used in the generation of human-mouse hybridomas that synthesize human monoclonal antibodies.