Functional heterogeneity of human tissue-resident memory T cells based on dye efflux capacities
Brahma V. Kumar, Radomir Kratchmarov, Michelle Miron, Dustin J. Carpenter, Takashi Senda, Harvey Lerner, Amy Friedman, Steven L. Reiner, Donna L. Farber
Brahma V. Kumar, Radomir Kratchmarov, Michelle Miron, Dustin J. Carpenter, Takashi Senda, Harvey Lerner, Amy Friedman, Steven L. Reiner, Donna L. Farber
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Research Article Immunology

Functional heterogeneity of human tissue-resident memory T cells based on dye efflux capacities

Abstract

Tissue-resident memory T cells (TRMs) accelerate pathogen clearance through rapid and enhanced functional responses in situ. TRMs are prevalent in diverse anatomic sites throughout the human lifespan, yet their phenotypic and functional diversity has not been fully described. Here, we identify subpopulations of human TRMs based on the ability to efflux fluorescent dyes [efflux(+) TRMs] located within mucosal and lymphoid sites with distinct transcriptional profiles, turnover, and functional capacities. Compared with efflux(–) TRMs, efflux(+) TRMs showed transcriptional and phenotypic features of quiescence including reduced turnover, decreased expression of exhaustion markers, and increased proliferative capacity and signaling in response to homeostatic cytokines. Moreover, upon activation, efflux(+) TRMs secreted lower levels of inflammatory cytokines such as IFN-γ and IL-2 and underwent reduced degranulation. Interestingly, analysis of TRM subsets following activation revealed that both efflux(+) and efflux(–) TRMs undergo extensive transcriptional changes following TCR ligation but retain core TRM transcriptional properties including retention markers, suggesting that TRMs carry out effector function in situ. Overall, our results suggest a model for tissue-resident immunity wherein heterogeneous subsets have differential capacities for longevity and effector function.

Authors

Brahma V. Kumar, Radomir Kratchmarov, Michelle Miron, Dustin J. Carpenter, Takashi Senda, Harvey Lerner, Amy Friedman, Steven L. Reiner, Donna L. Farber

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Figure 5

Efflux(+) TRM subset stability and function following stimulation.

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Efflux(+) TRM subset stability and function following stimulation. (A) S...
(A) Sorted efflux(+) and efflux(–) TRMs were stimulated with anti–CD3/CD28/CD2 beads, and efflux capacity was reassessed after 48 hours along with CD25 expression. Plots show MitoTracker and CD25 expression from the spleen of 1 donor for the indicated subsets. (B) Cytokine production following TCR stimulation. Efflux(+) and efflux(–) TRMs were sorted and stimulated with anti–CD3/CD28/CD2 beads for 72 hours, and cytokines in supernatant were quantified using cytometric bead array (see Methods). Graphs show levels of indicated cytokines in supernatants compiled from 8 donors. (C) Degranulation of efflux(+) and efflux(–) TRM subsets. Sorted cells were pulse labeled with CD107a antibody followed by PMA/ionomycin stimulation. Representative plots and quantification of CD107a+ from splenic efflux(+) and efflux(–) TRM subsets from 4 donors. For all panels, *P ≤ 0.05. **P ≤ 0.01 ****P ≤ 0.0001 by paired t test. ns, not significant.