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Identification and Isolation of Synovial Dendritic Cells

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Dendritic Cell Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 64))

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Abstract

In rheumatoid arthitis patients, three compartments need to be considered: peripheral blood (PB), synovial fluid (SF), and synovial tissue (ST). Dendritic cells (DC) characterized from each compartment have different properties. The methods given are based on cell sorting for isolation of cells, and flow cytometry and immunohistochemical staining for analysis of cells. Myeloid non-T cells are first enriched by density gradient centrifugation, sheep erythocyte rosetting, and, in some cases, magnetic immunodepletion. By flow cytometry, DC can then be analyzed or sorted based on two- or three-color immunofluorescence. Some variations on this basic theme are also outlined. The basic protocol for two-color immunohistochemistry of formalin-fixed ST is then given. This is based on the localization of the NFκB family member, RelB, to the nucleus of differentiated DC, and exclusion of B cells, macrophages, and follicular DC by double staining. Some variations that are particularly useful in frozen sections follow.

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Author information

Authors and Affiliations

  1. Centre for Immunology and Cancer Research, University of Queensland, Brisbane, Australia

    Allison R. Pettit, Lois Cavanagh & Ranjeny Thomas

Authors
  1. Allison R. Pettit
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  2. Lois Cavanagh
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  3. Ranjeny Thomas
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Editor information

Editors and Affiliations

  1. Department of Hematology, University College Hospital London, UK

    Stephen P. Robinson MD, PhD

  2. Antigen Presentation Research Group, Imperial College School of Medicine, at Northwick Park Hospital, Harrow, Middlesex, UK

    Andrew J. Stagg PhD

  3. Professor of Immunopathology, Imperial College School of Medicine, London

    Stella C. Knight PhD

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© 2001 Humana Press Inc.

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Pettit, A.R., Cavanagh, L., Thomas, R. (2001). Identification and Isolation of Synovial Dendritic Cells. In: Robinson, S.P., Stagg, A.J., Knight, S.C. (eds) Dendritic Cell Protocols. Methods in Molecular Medicine™, vol 64. Humana Press. https://doi.org/10.1385/1-59259-150-7:175

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  • DOI: https://doi.org/10.1385/1-59259-150-7:175

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-584-3

  • Online ISBN: 978-1-59259-150-3

  • eBook Packages: Springer Protocols

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