Characterization of a B cell helper factor(s) derived from CD5+ B cell hybridomas.
Work from our laboratory suggests that the selective advantage of frequently autoreactive CD5+ B cells is to provide activation signals to CD5- antigen-specific B cells. This hypothesis is supported by the observation that supernatants from CD5+ B cell hybridomas replace CD5+ B cell populations in helping idiotypic B cell subsets respond to antigen plus anti-idiotype antibody. The present study was designed to initiate the characterization of CD5+ B hybridoma-derived helper factor(s) (BHF) and to compare BHF to previously described cytokines. Elution of BHF from a lectin column enabled significant enrichment of the apparently glycosylated helper factor(s) from serum-free hybridoma supernatant. Gel filtration of this enriched activity revealed two significant peaks of helper activity, one at approximately 19-22 kDa and a second at 29-32 kDa. BHF activity in each fraction was sensitive to protease treatment. To determine if some previously described cytokines of approximately the same molecular weights were responsible for BHF activity, BHF fractions were tested for cytokine activity in respective bioassays. At least 2000 units of BHF did not contain detectable levels of IL-1, IL-2, IL-3, IL-4, IL-6, GM-CSF, G-CSF, or IFN-gamma activity. Furthermore, three hybridomas which produced BHF did not transcribe detectable levels of mRNAs specific for IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, GM-CSF, or IFN-gamma. The results suggest that CD5+ B cell hybridomas produce a lymphokine(s) distinct from cytokines commonly associated with B cell activation. The potential roles of this lymphokine in immunity and disease are discussed.
Harvard Medical School, Department of Pathology, Boston, Massachusetts 02115.
J A
JA
Hardin
M
M
Gibson
Y
Y
Kawano
S
S
Spielberg
C
C
Martin
M
M
Collins
M E
ME
Dorf
D H
DH
Sherr
D000818:Animals; D015703:Antigens, CD / Q000032:analysis*; D000943:Antigens, Differentiation / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D018956:CD5 Antigens; D003115:Colony-Stimulating Factors / Q000032:analysis / Q000096:biosynthesis; D006825:Hybridomas / Q000276:immunology*; D007130:Immunoglobulin Idiotypes / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis; D007378:Interleukins / Q000032:analysis / Q000096:biosynthesis; D008222:Lymphokines / Q000302:isolation & purification* / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D014158:Transcription, Genetic
D015703:Antigens, CD; D000943:Antigens, Differentiation; D018956:CD5 Antigens; D003115:Colony-Stimulating Factors; D007130:Immunoglobulin Idiotypes; D007378:Interleukins; D008222:Lymphokines; D007371:Interferon-gamma