Functional bioassays for B cell growth factors using polyclonally activated murine spleen B cells.
In this study, we have set up and optimized three distinct T cell-independent polyclonal B cell activation assay systems using highly T cell-depleted murine spleen B cells which were either preactivated in vitro with lipopolysaccharide (LPS) or costimulated with anti-IgM antibodies (anti-mu) or dextran sulfate (DxS). Using these assays, we have investigated the effects of recombinant human or murine interleukins, as well as those of a partially purified T cell-derived factor, designated BSF-LPS. Our results show that none of the interleukins, alone or in combination, is able to maintain growth of the LPS-preactivated B cell blasts, even at the highest concentrations tested, whereas the addition of our BSF-LPS preparation to the cultures significantly increases DNA synthesis. As expected, recombinant murine IL-4 (r mu IL-4) causes a substantial proliferation of anti-mu costimulated B cells. Such anti-mu costimulated B cells also respond, to a lower degree, to recombinant human IL-1 alpha (r hu IL-1 alpha), and do not significantly proliferate upon addition of r mu IL-2, r mu IL-5 or BSF-LPS. On the other hand, r mu IL-5 stimulates very efficiently DxS-costimulated B cells proliferation, whereas r hu IL-1 alpha only exerts a marginal effect; r mu IL-2, r mu IL-4 or BSF-LPS did not maintain growth of DxS-costimulated B cells. We believe that such a thorough investigation of the particular behaviour of activated B cell subpopulations towards various lymphokines provides the background for setting up a valuable bioassay method to differentiate the various interleukins acting on B cells through parallel use of the three distinct T cell-independent polyclonal B cell activation assay systems.
Ruuth
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INSERM U 25, Hôpital Necker, Paris, France.
Couillin
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Herbelin
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Praz
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D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000276:immunology*; D001681:Biological Assay; D002455:Cell Division; D002478:Cells, Cultured; D016264:Dextran Sulfate; D003911:Dextrans / Q000494:pharmacology; D007075:Immunoglobulin M / Q000276:immunology; D015847:Interleukin-4; D015848:Interleukin-5; D007378:Interleukins / Q000032:analysis* / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008934:Mitogens; D011994:Recombinant Proteins / Q000032:analysis; D013154:Spleen / Q000166:cytology / Q000276:immunology*
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D003911:Dextrans; D007075:Immunoglobulin M; D015848:Interleukin-5; D007378:Interleukins; D008070:Lipopolysaccharides; D008934:Mitogens; D011994:Recombinant Proteins; D015847:Interleukin-4; D016264:Dextran Sulfate