Epitope-specific CD4+ T cells can be labeled in complex cell mixtures from secondary lymphoid organs with fluorophore-labeled peptide:major histocompatibility complex class II (p:MHCII) tetramers and then detected by flow cytometry. Magnetic enrichment of tetramer-bound cells before flow cytometry increases the sensitivity of detection to the point where epitope-specific cells can be studied even when very rare at early and late times after the host has been exposed to the epitope. This method is very useful for studying polyclonal epitope-specific CD4+ T cells under physiological conditions.
Bibliographical noteFunding Information:
We thank J. Walter, C. Ellwood, and the University of Minnesota Flow Cytometry Resource for technical assistance. This work was supported by U. S. National Institutes of Health grants to M.K.J. (R01 AI039614 and P01 AI35296) and D.I.K. (T32 AI083196 and T32 AI007313).
© 2019 John Wiley & Sons, Inc.
- CD4 T cells
- epitope specific
- magnetic-bead enrichment
- naive CD4 T cells