IL-4 prevents adenosine-mediated immunoregulation by inhibiting CD39 expression
Fengqin Fang, Wenqiang Cao, Yunmei Mu, Hirohisa Okuyama, Lingjie Li, Jingtao Qiu, Cornelia M. Weyand, Jörg J. Goronzy
Fengqin Fang, Wenqiang Cao, Yunmei Mu, Hirohisa Okuyama, Lingjie Li, Jingtao Qiu, Cornelia M. Weyand, Jörg J. Goronzy
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Research Article Aging

IL-4 prevents adenosine-mediated immunoregulation by inhibiting CD39 expression

Abstract

The ectonucleotidase CD39 functions as a checkpoint in purinergic signaling on effector T cells. By depleting eATP and initiating the generation of adenosine, it impairs memory cell development and contributes to T cell exhaustion, thereby causing defective tumor immunity and deficient T cell responses in older adults who have increased CD39 expression. Tuning enzymatic activity of CD39 and targeting the transcriptional regulation of ENTPD1 can be used to modulate purinergic signaling. Here, we describe that STAT6 phosphorylation downstream of IL-4 signaling represses CD39 expression on activated T cells by inducing a transcription factor network including GATA3, GFI1, and YY1. GATA3 suppresses ENTPD1 transcription through prevention of RUNX3 recruitment to the ENTPD1 promoter. Conversely, pharmacological STAT6 inhibition decreases T cell effector functions via increased CD39 expression, resulting in the defective signaling of P2X receptors by ATP and stimulation of A2A receptors by adenosine. Our studies suggest that inhibiting the STAT6 pathway to increase CD39 expression has the potential to treat autoimmune disease while stimulation of the pathway could improve T cell immunity.

Authors

Fengqin Fang, Wenqiang Cao, Yunmei Mu, Hirohisa Okuyama, Lingjie Li, Jingtao Qiu, Cornelia M. Weyand, Jörg J. Goronzy

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Figure 2

STAT6 inhibition in a T cell response upregulates CD39 expression.

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STAT6 inhibition in a T cell response upregulates CD39 expression. (A) P...
(A) Purified total T cells were activated for 5 days by anti-CD3/CD28 Dynabeads. Each day, cells were examined for the expression of pSTAT6, GATA3, and CD39 in CD4 or CD8 T cells by flow cytometry. In addition, the level of pSTAT6 was also compared in T cells with or without IL-4 receptor blocking Abs. (B) Purified total T cells were activated for 3 days by anti-CD3/CD28 Dynabeads in the presence or absence of IL-4 (20, 50, or 100 ng/mL), IL-4 receptor blocking Abs (1 or 2 μg/mL), or STAT6 inhibitor AS1517499 (100 nM) as indicated. GATA3 expression was assessed by Western blotting. (C) Purified total T cells were activated in the presence or absence of the STAT6 inhibitor AS1517499 (100 nM). After 4 days, CD39 expression was evaluated by flow cytometry. (D) Total T cells were activated for 2 days before nucleofection with STAT6-specific siRNA smartpool. After 2 additional days, cells were collected for ENTPD1 and STAT6 mRNA quantification by real-time qPCR. (E) CD45.1+ naive SMARTA cells were isolated and injected into CD45.2 B6 WT mice via tail vein, followed by LCMV Armstrong infection and daily i.p. injection of the STAT6 inhibitor AS1517499 or DMSO solvent control. After 6 days, SMARTA cells were isolated from spleen and examined for CD39 expression using flow cytometry. Representative contour plots (upper panel) and summary data (lower panel). Data in D are shown as mean ± SEM and compared by 2-tailed paired or unpaired Student’s t test in C and E.