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Cell Differentiation in the Presence of Cytochalasin B: Studies on the "Switch" to IgG Secretion after Polyclonal B Cell Activation¹

From the Department of Genetics, and the Division of Immunology, Department of Medicine, Stanford University School of Medicine, Stanford, California 94305

Abstract

Mouse spleen cells were cultured with lipopolysaccharide in conditions that activate both IgM and IgG secretion. Addition of cytochalasin B (CB), an inhibitor of cytokinesis, lead to a high degree of polynucleation, with little effect on Ig secretion. Using cytoplasmic staining with fluorochrome conjugated antisera, we determined the numbers of IgG-containing cells that also contained IgM in their cytoplasm. Such double staining cells were relatively more frequent at early times of the cultures, but at all times single producing cells were in the majority. Addition of CB over the period when the IgG producing cells first appear, lead to a marked increased frequency of double staining, polynucleated cells. This characteristic was stable over a period of at least 42 hr, suggesting that each double staining cell actively synthesized both isotypes. When CB was added after IgG production had started, little increase in the numbers of double staining cells were observed, although polynucleation remained extensive. These data confirm previous findings that the lineage of one cell can produce both IgM and IgG. Furthermore, the results suggest that cells in the process of switching from IgM to IgG go through an asymmetric division leading to one IgM-producing and one IgG-producing daughter cell.

Footnotes

¹ This work was supported by grants from the National Institute of General Medical Sciences (GM 17367), National Cancer Institute (CA 04681), and National Institute of Allergy and Infectious Diseases (AI 10293).

² Present address: Vrije Universiteit Brussel, Instituut voor Molekulaire, Biologie, Paardenstraat 65, 1640 St. Genesius-Rode, Brussels, Belgium. Research Fellow, Belgium National Science Foundation (N. F. W. O.).

³ Postdoctoral Fellow, American Cancer Society, California Division. Correspondence should be addressed to: Dr. Eva Severinson Gronowicz, Department of Medicine (Immunology), Stanford University School of Medicine, Stanford, California 94305.

⁴ Investigator, Howard Hughes Medical Institute.

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