Glycolysis determines dichotomous regulation of T cell subsets in hypoxia
Yang Xu, Arindam Chaudhury, Ming Zhang, Barbara Savoldo, Leonid S. Metelitsa, John Rodgers, Jason T. Yustein, Joel R. Neilson, Gianpietro Dotti
Yang Xu, Arindam Chaudhury, Ming Zhang, Barbara Savoldo, Leonid S. Metelitsa, John Rodgers, Jason T. Yustein, Joel R. Neilson, Gianpietro Dotti
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Research Article Immunology

Glycolysis determines dichotomous regulation of T cell subsets in hypoxia

Abstract

Hypoxia occurs in many pathological conditions, including chronic inflammation and tumors, and is considered to be an inhibitor of T cell function. However, robust T cell responses occur at many hypoxic inflammatory sites, suggesting that functions of some subsets are stimulated under low oxygen conditions. Here, we investigated how hypoxic conditions influence human T cell functions and found that, in contrast to naive and central memory T cells (TN and TCM), hypoxia enhances the proliferation, viability, and cytotoxic action of effector memory T cells (TEM). Enhanced TEM expansion in hypoxia corresponded to high hypoxia-inducible factor 1α (HIF1α) expression and glycolytic activity compared with that observed in TN and TCM. We determined that the glycolytic enzyme GAPDH negatively regulates HIF1A expression by binding to adenylate-uridylate–rich elements in the 3′-UTR region of HIF1A mRNA in glycolytically inactive TN and TCM. Conversely, active glycolysis with decreased GAPDH availability in TEM resulted in elevated HIF1α expression. Furthermore, GAPDH overexpression reduced HIF1α expression and impaired proliferation and survival of T cells in hypoxia, indicating that high glycolytic metabolism drives increases in HIF1α to enhance TEM function during hypoxia. This work demonstrates that glycolytic metabolism regulates the translation of HIF1A to determine T cell responses to hypoxia and implicates GAPDH as a potential mechanism for controlling T cell function in peripheral tissue.

Authors

Yang Xu, Arindam Chaudhury, Ming Zhang, Barbara Savoldo, Leonid S. Metelitsa, John Rodgers, Jason T. Yustein, Joel R. Neilson, Gianpietro Dotti

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Figure 6

The expression of HIF1α in T cell memory subsets is translationally regulated by GAPDH.

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The expression of HIF1α in T cell memory subsets is translationally regu...
(A) A GFP/HIF1A 3′-UTR reporter construct was introduced into PB-Ts and TEXP by nucleofection, and GFP expression was measured 8 hours after transfection. The percentages in A indicate the percentage of GFP+ cells. n = 3 for PB-Ts and n = 3 for TEXP. *P = 0.03, paired Student’s t test. (B) GAPDH-specific Abs were used to immunoprecipitate GAPDH from PB-Ts and TEXP. Mouse IgG Ab was used as a control for the IP. The immunoprecipitated mRNA was analyzed by qPCR to quantify HIF1A and ACTB mRNA. Data were normalized against the IgG control pulldown. n = 3. **P = 0.0081, 2-way ANOVA with Bonferroni’s post-hoc analysis. (C) Quantification of HIF1α protein expression in mock-transduced TEXP or GAPDH-transduced (GAPDH-Td) TEXP that were unstimulated or stimulated with OKT3/a-CD28 Abs in normoxia or hypoxia. n = 3. ****P < 0.0001, 2-way ANOVA with Bonferroni’s post-hoc analysis. (D) Cell counts and cell viability were determined in the TEXP 72 hours after activation. n = 4. *P < 0.05 and ***P = 0.0002, 2-way ANOVA with Bonferroni’s post-hoc analysis. Error bars indicate SD. Dotted line indicates the starting cell number.