Functional and biochemical characterization of B-cell differentiation factor (BCDF) produced by an HTLV-I-transformed human T-cell clone and demonstration of specific binding of the factor to a BCDF responsive cell line.
We have previously described YA2, a human T-cell clone that secretes B-cell differentiation factor (BCDF) but not B-cell growth factor (BCGF). The BCDFs secreted by YA2 and HTLV-I-transformed YA2 (TYA2) were functionally similar in their ability to stimulate Ig secretion by Staphylococcus aureus Cowan strain I-activated B cells and IgM secretion by SKW6.4 cells. In addition, they were biochemically similar with a MW of 30 kDa by high-performance liquid chromatography (HPLC) sieving, and a pI of 6.0-6.8 by isoelectric focusing. The BCDF activity was not blocked by antibodies to interleukin 2 and BCGF. BCDF was purified from TYA2 supernatant by sequential media protein immunoadsorption, flat bed isoelectric focusing, HPLC TSK 2000 sieving, and repeated immunoadsorption and was then iodinated. The iodinated material had functional BCDF activity and migrated to a single band at MW 30 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and at pI of 6.8 by polyacrylamide gel isoelectric focusing. 125I-BCDF purified in this manner bound specifically to a BCDF-responsive cell line and not to phytohemagglutinin-activated T cells. 125I binding to the BCDF-responsive cell line was competitively inhibitable by the addition of cold BCDF. Thus we have purified and characterized a factor with BCDF activity and demonstrated that this factor binds specifically to a BCDF-responsive cell line.
D000917:Antibody Formation; D001402:B-Lymphocytes / Q000276:immunology*; D001665:Binding Sites; D002460:Cell Line; D002472:Cell Transformation, Viral; D017977:Deltaretrovirus / Q000276:immunology; D006800:Deltaretrovirus Infections / Q000276:immunology*; D006133:Growth Substances / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D007376:Interleukin-2 / Q000502:physiology; D015847:Interleukin-4; D007526:Isoelectric Point; D008222:Lymphokines / Q000302:isolation & purification / Q000502:physiology*; D008970:Molecular Weight; D013601:T-Lymphocytes / Q000276:immunology*
D006133:Growth Substances; D007376:Interleukin-2; D008222:Lymphokines; D015847:Interleukin-4