Tracking by Flow Cytometry Antigen-Specific Follicular Helper T Cells in Wild-Type Animals After Protein Vaccination
Flow cytometry is a valuable technology used in immunology to characterize and enumerate the different cell subpopulations specific for a nonself-antigen in the context of an ongoing immune response. Among them, follicular helper T cells are the cognate regulators of B cells in secondary lymphoid tissues. Thus, tracking them is of high interest especially in the context of protein vaccination. For this purpose, transgenic antigen-receptor mouse models have been largely used. It is now clear that transgenic models are not always the best means to study the dynamics of the immune response since they can modify the response. In this chapter, we describe how to track endogenous antigen-specific follicular helper T cells by flow cytometry after protein vaccination in nonmodified wild-type animals, which ultimately provides a comprehensive way to enumerate, characterize, and isolate these particular cells in vivo.
Key wordsFlow cytometry pMHC II tetramer Monoclonal antibodies T lymphocytes B-cell help
This work was supported by AVENIR INSERM, Association pour la Recherche sur le Cancer, la Ligue Nationale Contre le Cancer, Conseil Régional Midi-Pyrénées, Institut National contre le Cancer (INCa, PLBIO10-195 and INCA-6530), and International Reintegration Grant Marie Curie to NF.