Sprouty-2 regulates HIV-specific T cell polyfunctionality
Yen-Ling Chiu, … , Diane E. Griffin, Jonathan P. Schneck
Yen-Ling Chiu, … , Diane E. Griffin, Jonathan P. Schneck
Published December 2, 2013
Citation Information: J Clin Invest. 2014;124(1):198-208. https://doi.org/10.1172/JCI70510.
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Research Article Virology

Sprouty-2 regulates HIV-specific T cell polyfunctionality

Abstract

The ability of individual T cells to perform multiple effector functions is crucial for protective immunity against viruses and cancer. This polyfunctionality is frequently lost during chronic infections; however, the molecular mechanisms driving T cell polyfunctionality are poorly understood. We found that human T cells stimulated by a high concentration of antigen lacked polyfunctionality and expressed a transcription profile similar to that of exhausted T cells. One specific pathway implicated by the transcription profile in control of T cell polyfunctionality was the MAPK/ERK pathway. This pathway was altered in response to different antigen concentrations, and polyfunctionality correlated with upregulation of phosphorylated ERK. T cells that were stimulated with a high concentration of antigen upregulated sprouty-2 (SPRY2), a negative regulator of the MAPK/ERK pathway. The clinical relevance of SPRY2 was confirmed by examining SPRY2 expression in HIV-specific T cells, where high levels of SPRY2 were seen in HIV-specific T cells and inhibition of SPRY2 expression enhanced the HIV-specific polyfunctional response independently of the PD-1 pathway. Our findings indicate that increased SPRY2 expression during chronic viral infection reduces T cell polyfunctionality and identify SPRY2 as a potential target for immunotherapy.

Authors

Yen-Ling Chiu, Liang Shan, Hailiang Huang, Carl Haupt, Catherine Bessell, David H. Canaday, Hao Zhang, Ya-Chi Ho, Jonathan D. Powell, Mathias Oelke, Joseph B. Margolick, Joel N. Blankson, Diane E. Griffin, Jonathan P. Schneck

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Figure 6

SPRY2 inhibition enhances HIV-specific T cell polyfunctionality.

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SPRY2 inhibition enhances HIV-specific T cell polyfunctionality. (A a...
(A and B) SPRY2 expression was studied in 12 HIV-infected HLA-A2+ patients. HIV-Gag-specific T cells and influenza M1-specific T cells were sorted by pentamer/tetramer staining and mRNA extracted from Gag- and M1-specific cells for qPCR analysis. (A) An example of flow cytometry–based simultaneous analysis of Gag- and M1-specific T cells from PBMCs of an HIV-infected donor. (B) qPCR and flow cytometry analysis of SPRY2 and PD-1 expression in Gag-specific, M1-specific, and total CD8+ T cells from HLA-A2+ HIV donors. Statistical analyses were performed by nonparametric Wilcoxon matched-pairs signed rank test. (C–F) PBMCs from 19 HIV-infected patients were activated with soluble anti-CD3, anti-CD28, and a mix of CEF/HIV peptide pools. Some cultures were also treated with anti–PD-1 (10 μg/ml) to block PD-1 signaling during activation. 24 hours later, cells were transduced with SPRY2 knockdown (KD) or NT control lentivirus. On D7 after virus transduction, PBMCs were stimulated with CEF or HIV Gag, Nef, and Tat peptide pools for 6 hours and analyzed for polyfunctionality. (C–E) Inhibition of SPRY2 expression led to augmented HIV-specific T cell polyfunctionality above and beyond that seen by inhibition of PD-1 pathway alone. (F) PD-1 blockade alone or in combination with SPRY2 inhibition had no significant effect on the CEF-specific response.