IL-10–producing NKT10 cells are a distinct regulatory invariant NKT cell subset
Duygu Sag, … , Mitchell Kronenberg, Gerhard Wingender
Duygu Sag, … , Mitchell Kronenberg, Gerhard Wingender
Published July 25, 2014
Citation Information: J Clin Invest. 2014;124(9):3725-3740. https://doi.org/10.1172/JCI72308.
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Research Article Immunology

IL-10–producing NKT10 cells are a distinct regulatory invariant NKT cell subset

Abstract

Invariant natural killer T (iNKT) cells rapidly produce copious amounts of multiple cytokines after activation, thereby impacting a wide variety of different immune reactions. However, strong activation of iNKT cells with α-galactosylceramide (αGalCer) reportedly induces a hyporeactive state that resembles anergy. In contrast, we determined here that iNKT cells from mice pretreated with αGalCer retain cytotoxic activity and maintain the ability to respond to TCR-dependent as well as TCR-independent cytokine-mediated stimulation. Additionally, αGalCer-pretreated iNKT cells acquired characteristics of regulatory cells, including production and secretion of the immunomodulatory cytokine IL-10. Through the production of IL-10, αGalCer-pretreated iNKT cells impaired antitumor responses and reduced disease in experimental autoimmune encephalomyelitis, a mouse model of autoimmune disease. Furthermore, a subset of iNKT cells with a similar inhibitory phenotype and function were present in mice not exposed to αGalCer and were enriched in mouse adipose tissue and detectable in human PBMCs. These data demonstrate that IL-10–producing iNKT cells with regulatory potential (NKT10 cells) represent a distinct iNKT cell subset.

Authors

Duygu Sag, Petra Krause, Catherine C. Hedrick, Mitchell Kronenberg, Gerhard Wingender

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Figure 9

A subset of untreated Vα14i NKT cells express IL-10.

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A subset of untreated Vα14i NKT cells express IL-10. (A and B) C57BL/6 m...
(A and B) C57BL/6 mice were left untreated or were injected with 1 μg αGalCer, and 90 minutes later, splenocytes were purified and incubated for 2 hours in vitro in the presence of protein transport inhibitors before iNKT cells were analyzed (αGalCer). Alternatively, splenocytes were stimulated with PMA and ionomycin in vitro for 4 hours (PMA + ionomycin). Representative data (A) and a summary graph (B) are shown. Data in B are summarized from at least 4 independent experiments with at least 9 mice per group. αGalCer = mean: 0.45% ± 0.03%; median: 0.46%. PMA + ionomycin = mean: 1.77% ± 0.18%; median: 1.55%. Numbers in the dot plots denote the percentage of cells in the respective rectangle. Gating is based on 0% of IL-10+ cells in unstimulated C57BL/6 iNKT cells. Similar background values were obtained with isotype control antibody or by staining of stimulated Il10–/– iNKT cells (Supplemental Figure 8). (C) Splenocytes from C57BL/6 mice were stimulated with PMA and ionomycin as described in A, and iNKT cells were stained for IL-10 and the indicated markers. Histograms depict the overlay of IL-10– (gray) and IL-10+ (black) iNKT cells for the indicated markers. Numbers below the graphs represent either the average expression of the indicated marker on IL-10– (gray) and IL-10+ (black) iNKT cells (left), or the fold difference between both of these values (right). All differences were statistically significant (P < 0.01). Data shown are representative of 3 to 6 independent experiments.