Establishment of rat-mouse T cell hybridomas that constitutively produce a soluble factor that is needed for the generation of cytotoxic cells: biochemical and functional characterization.
Spleen cells from Lewis rats were cultured with 4 micrograms/ml Con A. These cells were then fused with BW 5147 mouse T lymphoma cells. Two hybrid clones (6B2-B8 and 6B2-E6) obtained by fusion formed CGF effectively. It was found that hybrid cells can be boosted to produce higher levels of CGF upon stimulation with Con A. 6B2-B8 express rat T cell markers. CGF formed by 6B2-B8 had a m.w. of 23,000 and 40,000. CGF was eluted from a Mono Q anion-exchange column with an FPLC system at 0.4 to 0.6 M NaCl as a major peak and at 0.8 M NaCl as a minor peak. CGF was eluted as three peaks with pH 4.1, 4.8, and 5.2 from a Mono P chromatofocusing column. CGF from 6B2-B8 does not contain IL-1, IL-2, IL-3, or CSF.
D000818:Animals; D002459:Cell Fusion / Q000379:methods; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003602:Cytotoxicity, Immunologic; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D007621:Karyotyping; D055646:Killer Factors, Yeast; D008213:Lymphocyte Activation; D051379:Mice; D008806:Mice, Inbred AKR; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D055646:Killer Factors, Yeast; D011506:Proteins