The use of supernatants from neuraminidase and galactose oxidase (NAGO)-treated lymphocytes as a source of T cell growth factor.
Human peripheral blood or tonsil lymphocytes produce T cell growth factor (TCGF), when activated with neuraminidase (NA) and galactose oxidase (GO). Partial purification of NAGO-TCGF on Sepharose G-100 columns gave a TCGF-active fraction within the same molecular weight range as the conventional lectin-induced TCGF (approximately 15,000 Da). Human T cells, activated in mixed lymphocyte culture (MLC) with irradiated allogeneic EB-virus transformed B-cells (LCL) could be maintained in continuous culture for several months with retained functional activities. The cells showed similar growth patterns when cultured in the presence of either NAGO-TCGF or PHA-TCGF. The growing cells were characterized by means of monoclonal antibodies. After 4 weeks of culture 98% of these were OKT3+ and 87% were also OKT8+. The cytolytic activities of the cultures were tested in cell-mediated lympholysis (CML) against allogeneic LCL as target cells, in natural cytotoxicity (NK) against K562 cells and in antibody dependent cytotoxicity (ADCC) against bovine erythrocytes. Cultures displaying one or several of these functions were obtained. The results indicate, that TCGF obtained from supernatants of NAGO-activated lymphocytes is as potent as the T cell growth promoting factor obtained by lectin stimulation. One major advantage of using NAGO-generated TCGF is that contamination with lectin is avoided.
D000954:Antigens, Surface / Q000032:analysis; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D005692:Galactose Oxidase / Q000494:pharmacology; D006801:Humans; D007111:Immunity, Cellular; D007376:Interleukin-2 / Q000302:isolation & purification*; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology
D000954:Antigens, Surface; D007376:Interleukin-2; D005692:Galactose Oxidase; D009439:Neuraminidase