ABC transporters and NR4A1 identify a quiescent subset of tissue-resident memory T cells
Chandra Sekhar Boddupalli, Shiny Nair, Simon M. Gray, Heba N. Nowyhed, Rakesh Verma, Joanna A. Gibson, Clara Abraham, Deepak Narayan, Juan Vasquez, Catherine C. Hedrick, Richard A. Flavell, Kavita M. Dhodapkar, Susan M. Kaech, Madhav V. Dhodapkar
Chandra Sekhar Boddupalli, Shiny Nair, Simon M. Gray, Heba N. Nowyhed, Rakesh Verma, Joanna A. Gibson, Clara Abraham, Deepak Narayan, Juan Vasquez, Catherine C. Hedrick, Richard A. Flavell, Kavita M. Dhodapkar, Susan M. Kaech, Madhav V. Dhodapkar
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Research Article Immunology

ABC transporters and NR4A1 identify a quiescent subset of tissue-resident memory T cells

Abstract

Immune surveillance in tissues is mediated by a long-lived subset of tissue-resident memory T cells (Trm cells). A putative subset of tissue-resident long-lived stem cells is characterized by the ability to efflux Hoechst dyes and is referred to as side population (SP) cells. Here, we have characterized a subset of SP T cells (Tsp cells) that exhibit a quiescent (G0) phenotype in humans and mice. Human Trm cells in the gut and BM were enriched in Tsp cells that were predominantly in the G0 stage of the cell cycle. Moreover, in histone 2B-GFP mice, the 2B-GFP label was retained in Tsp cells, indicative of a slow-cycling phenotype. Human Tsp cells displayed a distinct gene-expression profile that was enriched for genes overexpressed in Trm cells. In mice, proteins encoded by Tsp signature genes, including nuclear receptor subfamily 4 group A member 1 (NR4A1) and ATP-binding cassette (ABC) transporters, influenced the function and differentiation of Trm cells. Responses to adoptive transfer of human Tsp cells into immune-deficient mice and plerixafor therapy suggested that human Tsp cell mobilization could be manipulated as a potential cellular therapy. These data identify a distinct subset of human T cells with a quiescent/slow-cycling phenotype, propensity for tissue enrichment, and potential to mobilize into circulation, which may be harnessed for adoptive cellular therapy.

Authors

Chandra Sekhar Boddupalli, Shiny Nair, Simon M. Gray, Heba N. Nowyhed, Rakesh Verma, Joanna A. Gibson, Clara Abraham, Deepak Narayan, Juan Vasquez, Catherine C. Hedrick, Richard A. Flavell, Kavita M. Dhodapkar, Susan M. Kaech, Madhav V. Dhodapkar

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Figure 3

Cell-cycle analysis of Tsp cells.

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Cell-cycle analysis of Tsp cells. (A) Hoechst–Pyronin Y cell-cycle analy...
(A) Hoechst–Pyronin Y cell-cycle analysis on Tsp and NSP fraction from human blood CD8+ T cells (top panel), IEL (middle panel), and skin CD8+ Trm cells (bottom panel). Result is representative of 3 independent experiments. (B) Bar graph represents combined data of 3 independent experiments. (C) Bar graph displays percentage of G0 and G1 fractions in CD8+ Tsp and NSP gated on Trm compartment from PP, IEL in mice at day 35 (p.i.) with LCMV-Arm. (D) Schematic diagram showing the protocol for measuring GFP label retention in H2B-GFP mice upon LCMV-Arm infection. (E) FACS analysis on memory CD8+ T cells from BM, spleen, liver, IEL, and PP after 5 weeks of chase; the same is documented in the bar graph (n = 4–5 mice). (F) FACS analysis of GFP label retention in SP and NSP CD8+ Trm compartment at 5 weeks chase in H2B GFP mice infected with LCMV-Arm. (G) Graph represents percentages of GFP+ cells in SP and NSP CD8+ Trm cells. *P < 0.05. Mouse experiments were repeated twice; graphs represent analysis from 1 experiment (n = 4 mice).