Differential activity of recombinant lymphokines on mouse B cell proliferation and cell cycle progression are revealed by 5-bromo-2'-deoxyuridine/Hoechst 33258 dye flow cytometry.
Activation of resting mouse B cells with anti-mu chain antibodies (anti-mu) leads to cell proliferation. We have investigated the effect of recombinant T cell interleukins (IL 2 to IL 6) on such anti-mu-induced proliferation. No proliferative response was detected when IL 2, IL 3 and IL 6, either alone or in combination with anti-mu, were studied. Furthermore, neither IL 4 nor IL 5 could induce proliferation when added alone to B cell cultures. However, when combined with anti-mu, IL 4 as well as IL 5 stimulated cell growth. Analysis by 5-bromo-2'-deoxyuridine/Hoechst 33258 flow cytometry revealed distinct effects of IL 4 and IL 5 on B cell growth. In the presence of anti-mu, both IL 4 and IL 5 co-stimulated unfractionated splenic B cells. However, when B cells were separated into subpopulations by density, IL 4 proved to be a cell cycle progression factor, stimulating the majority of resting B cells to enter the cell cycle. In contrast, IL 5 had little effect on the resting fraction of B cells. Rather, IL 5 acted as a co-competence factor, stimulating predominantly low-density B cells. Following exposure of anti-mu alone, most B cells accumulated in the G1 of the second cycle. Upon addition of IL 4, the cells acquired the ability to progress into the next S phase compartment. Contrary to what is seen when B cells are stimulated by other mitogens, very few cells are in the G2 compartments after anti-mu plus IL 4 stimulation. This phenomenon was not due to a differential cell cycle progression rate. Our findings provide an analytical basis for fractionating cell-cycle-compartment-specific B cells for their molecular study.
Seyschab
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Department of Human Genetics, University of Würzburg.
Hoehn
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Rabinovitch
P S
PS
Chen
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D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D005434:Flow Cytometry; D007148:Immunoglobulin mu-Chains / Q000276:immunology; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D015848:Interleukin-5 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D011994:Recombinant Proteins / Q000494:pharmacology
D000888:Antibodies, Anti-Idiotypic; D007148:Immunoglobulin mu-Chains; D007376:Interleukin-2; D007377:Interleukin-3; D015848:Interleukin-5; D015850:Interleukin-6; D008222:Lymphokines; D011994:Recombinant Proteins; D015847:Interleukin-4