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Purification and Immune Phenotyping of B-1 Cells from Body Cavities of Mice

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Regulatory B Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2270))

Abstract

B-1 cells are fetal-origin B lymphocytes with unique developmental and functional characteristics that can generate natural, polyreactive antibodies with important functions in tissue homeostasis and immune defense. While B-1 cell frequencies in bone marrow and secondary lymphoid tissues are low, relative high frequencies exist within peritoneal and pleural cavities of mice, including both CD5+ and CD5− B-1 cells. These cells represent B-1 reservoirs that, when activated, migrate to lymphoid tissues to secrete antibodies and/or cytokines. Here, we outline efficient methods for the extraction and magnetic isolation of CD5+ B-1 cells from the peritoneal and pleural cavities as well as the separation and phenotypic characterization of CD5+ and CD5− B-1 cells by flow cytometry.

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Acknowledgments

Data for the B-1 cell FACS-sorts were kindly provided by Dr. Elizabeth E. Waffarn (Columbia University). This work was supported in part by NIH U19AI109962 and AI117890.

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Authors and Affiliations

  1. Center for Immunology & Infectious Diseases, Schools of Medicine and Veterinary Medicine, University of California, Davis, CA, USA

    Vanessa Yenson & Nicole Baumgarth

  2. University of Technology Sydney, Sydney, NSW, Australia

    Vanessa Yenson

  3. Dept. Pathology, Microbiology and Immunology, Schools of Medicine and Veterinary Medicine, University of California, Davis, CA, USA

    Nicole Baumgarth

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  1. Vanessa Yenson
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  2. Nicole Baumgarth
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Correspondence to Nicole Baumgarth .

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Editors and Affiliations

  1. Department of Medicine, University of Udine, Udine, Italy

    Francesca Mion

  2. Department of Medicine, University of Udine, Udine, Italy

    Silvia Tonon

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Yenson, V., Baumgarth, N. (2021). Purification and Immune Phenotyping of B-1 Cells from Body Cavities of Mice. In: Mion, F., Tonon, S. (eds) Regulatory B Cells. Methods in Molecular Biology, vol 2270. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1237-8_2

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  • DOI: https://doi.org/10.1007/978-1-0716-1237-8_2

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  • Print ISBN: 978-1-0716-1236-1

  • Online ISBN: 978-1-0716-1237-8

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