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Purification and Characterization of Murine MZ and T2-MZP Cells

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Regulatory B Cells

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

Abstract

The spleen is the second major reservoir of B cells in the adult. In the spleen, cells, generated in the bone marrow, are selected, mature, and become part of the peripheral B-cell pool. Murine spleen comprises several B-cell subsets representing various maturation stages and/or cell functions. The spleen is a complex lymphoid organ organized into two main structures with different functions: the red and white pulp. The red pulp is flowed with blood while the white pulp is organized in primary follicles, with a B-cell area composed of follicular B cells and a T-cell area surrounding a periarterial lymphatic sheath. The frontier between the red and white pulp is defined as the marginal zone (MZ) and contains the MZ B cells. Because B cells, localized in different areas, are characterized by distinct expression levels of B-cell receptor (BCR) and of other surface markers, splenic B-cell subsets can be easily identified and purified by flow cytometry analyses and fluorescence-activated cell sorting (FACS).

Here, we will focus on MZ B cells and on their precursors, giving some experimental hints to identify, generate, and isolate these cells. We will combine the use of FACS analysis and confocal microscopy to visualize MZ B cells in cell suspensions and in tissue sections, respectively. We will also give some clues to analyze B-cell repertoire on isolated MZ-B cells.

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Acknowledgments

We thank Dr. Claudio Pioli for his critical review of the manuscript.

Author information

Authors and Affiliations

  1. B Cell Physiopathology Unit, Immunology Research Area, Bambino Gesù Children Hospital, Rome, Italy

    M. Manuela Rosado, Marco Scarsella, Simona Cascioli, Ezio Giorda & Rita Carsetti

  2. Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden

    Alaitz Aranburu

Authors
  1. M. Manuela Rosado
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  2. Alaitz Aranburu
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  3. Marco Scarsella
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  4. Simona Cascioli
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  5. Ezio Giorda
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  6. Rita Carsetti
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Corresponding author

Correspondence to M. Manuela Rosado .

Editor information

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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Rosado, M.M., Aranburu, A., Scarsella, M., Cascioli, S., Giorda, E., Carsetti, R. (2021). Purification and Characterization of Murine MZ and T2-MZP Cells. 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_1

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  • DOI: https://doi.org/10.1007/978-1-0716-1237-8_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1236-1

  • Online ISBN: 978-1-0716-1237-8

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