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Identifying Regulatory B Cells (B10 Cells) that Produce IL-10 in Mice

  • Takashi Matsushita
  • Thomas F. Tedder
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 677)

Abstract

Regulatory B cells that produce IL-10 are now recognized as an important component of the immune system. We have identified a rare antigen-specific regulatory B-cell subset with a unique CD1dhiCD5+CD19hi phenotype in the spleens of wild-type mice. We call these cells B10 cells because they are responsible for most B cell IL-10 production, they appear to only produce IL-10 after 5 h of in vitro stimulation, and to distinguish them from other potential regulatory B cell subsets. B10 progenitor (B10pro) cells have also been identified within the spleen CD1dhiCD5+CD19hi B-cell subset, and within other lymphoid tissues. Herein, four methods for identifying and isolating regulatory IL-10-producing B10 cells in mice are provided. The first two methods are used to identify and enumerate B10 and B10pro cells based on their cell surface phenotypes and cytoplasmic IL-10 staining. The last two methods are used to isolate viable B10 cells for adoptive transfer and functional studies. These methods should facilitate the study of B10 cells in inflammation, autoimmune disease, immune responses, and cancer therapy.

Key words

B cells Autoimmunity Inflammation Regulatory cells CD5 Interleukin-10 

Notes

Acknowledgments

This work was supported by grants from the National Institutes of Health CA105001, CA96547, AI56363, and AI057157. We thank Drs. Guang Yang, Regina Lin, Kathleen Candando, and David DiLillo for their input and review of this manuscript.

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Copyright information

© Humana Press 2010

Authors and Affiliations

  • Takashi Matsushita
    • 1
  • Thomas F. Tedder
    • 1
  1. 1.Department of ImmunologyDuke University Medical CenterDurhamUSA

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