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The Journal of Immunology, 2006, 177: 1581-1589.
Copyright © 2006 by The American Association of Immunologists

Kinetics of B Cell Receptor Signaling in Human B Cell Subsets Mapped by Phosphospecific Flow Cytometry1

Jonathan M. Irish*,{dagger}, Debra K. Czerwinski*, Garry P. Nolan3,{dagger} and Ronald Levy2,3,*

* Department of Medicine, Oncology Division, Stanford University, Stanford, CA 94305; and {dagger} Department of Microbiology and Immunology, Baxter Laboratories for Genetic Pharmacology, Stanford University, Stanford, CA 94305

Differences in BCR signaling may govern outcomes as diverse as proliferation and cell death. We profiled BCR signaling kinetics in subsets of primary human B cells using flow cytometry. In the predominant population expressing IgM, BCR cross-linking led to a quick burst of Syk, ERK1/2, and p38 signaling. In contrast, IgG B cells sustained higher per-cell ERK1/2 phosphorylation over time. This dichotomy suggested a mechanism for dampening signals transmitted by IgM. Regulatory phosphatase activity in IgM B cells was BCR-mediated and initiated more slowly than kinase activity. This BCR-mediated phosphatase activity was sensitive to inhibition by H2O2 and required to attenuate IgM BCR signaling. These results provide the first kinetic maps of BCR signaling in primary human B cell subsets and enable new studies of signaling in B cell disorders, such as autoimmunity and cancer.




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