In vitro stimulated lymphocytes as a source of human hybridomas

Abstract

Stimulation of peripheral blood lymphocyte (PBL) cultures from tetanus toxoid (TT)-immunized donor with Epstein-Barr virus (EBV) yielded cells with much higher frequencies of hybrid formation (36 × 10⁻⁷) compared to unstimulated PBL or cells cultured with pokeweed mitogen or TT antigen. The proportion of hybridomas (approximately 1%) producing anti-TT antibody was similar in EBV- and TT-stimulated cultures. A marked increase in immunoglobulin secretion was observed after hybridization and preselection of EBV subcultures for high anti-TT production prior to fusion resulted in a fivefold increase in TT-specific hybridomas (pin vitro, with EBV, prior to fusion, should yield an increasing spectrum of human monoclonal antibodies for diagnostic, therapeutic or basic studies.