Protocol for phenotyping and isolation of dendritic cell subsets from blood and lymphoid organs of non-human primates and humans by flow cytometry
Dendritic cells (DCs) encompass several subsets that are essential for shaping immune responses. Here, we present a protocol for ex vivo functional analysis and purification of DC subsets from blood and secondary lymphoid organs using flow cytometry. We describe the steps for isolating mononuclear cells from blood, lymph nodes, and spleen. We then detail the procedures for phenotypic characterization of DC subsets, intracellular staining to assess their cytokine production, and fluorescence-activated cell sorting (FACS) to isolate individual DC populations. For complete details on the use and execution of this protocol, please refer to Gardet et al.<sup>1</sup>.
Authors
- Francis Relouzat (ORCID: https://orcid.org/0000-0002-6387-8481)
- Margaux Gardet (ORCID: https://orcid.org/0009-0000-0226-7022)
- J.A. Pons (ORCID: https://orcid.org/0000-0001-5017-7240)
- Florian Meurisse (ORCID: https://orcid.org/0009-0007-8239-7011)
- Olivier Lambotte (ORCID: https://orcid.org/0000-0003-4425-8516)
- Benoı̂t Favier (ORCID: https://orcid.org/0000-0001-9191-6273)
Institutions
- Inserm (FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- Université Paris-Saclay (FR)
- CEA Paris-Saclay (FR)
- Infectious Disease Models and Innovative Therapies (FR)
- Bicêtre Hospital (FR)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-06-13
- DOI
- https://doi.org/10.1016/j.xpro.2025.103897
- Primary Topic
- Immunotherapy and Immune Responses
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Agence Nationale de la Recherche
- Agence Nationale de Recherches sur le Sida et les Hépatites Virales