Characterization of monoclonal B cell growth factor (BCGF) produced by a human T-T hybridoma.
We previously demonstrated the development of a cloned human T cell hybridoma that secretes B cell growth factor (BCGF) in the absence of demonstrable interleukin 2 or B cell differentiation factor. Sephadex gel filtration chromatography demonstrated the m.w. of this factor to be 18 to 20K. The present studies were performed to further characterize the biochemical properties of the molecule and to determine its target cell specificity. Temperature stability studies showed the monoclonal BCGF to be stable at 37 degrees C for 12 hr and at 70 degrees C for 15 min; however, most (93%) of the activity was lost after incubation at 70 degrees C for 30 min. Aliquots of hybridoma supernatant were exposed to buffer solutions with variable pH with no diminution in activity over a pH range of 4.0 to 10.0 BCGF activity was not affected by 2-mercaptoethanol, neuraminidase, or nucleic acid denaturing enzymes. In contrast, all activity was destroyed by 10 M urea, trypsin, and chymotrypsin. Chromatofocusing demonstrated the isoelectric point of BCGF to be 6.3 to 6.6. Finally, absorption experiments demonstrated that BCGF activity was absorbed by large, activated B cells. Mitogen-stimulated T cell blasts, small resting B cells, and CESS cells failed to absorb BCGF activity from the hybridoma supernatant. These and future studies with purified monoclonal human BCGF should enhance our understanding of its immunochemical properties and of its role in the immunoregulation of human B cell responses.
D000042:Absorption; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D004355:Drug Stability; D006133:Growth Substances / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D015847:Interleukin-4; D007525:Isoelectric Focusing; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D010447:Peptide Hydrolases / Q000494:pharmacology; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D013696:Temperature
D006133:Growth Substances; D008222:Lymphokines; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D015847:Interleukin-4; D010447:Peptide Hydrolases