Granzyme A–producing T helper cells are critical for acute graft-versus-host disease
Sungtae Park, Brad Griesenauer, Hua Jiang, Djamilatou Adom, Pegah Mehrpouya-Bahrami, Srishti Chakravorty, Majid Kazemian, Tanbeena Imam, Rajneesh Srivastava, Tristan A. Hayes, Julian Pardo, Sarath Chandra Janga, Sophie Paczesny, Mark H. Kaplan, Matthew R. Olson
Sungtae Park, Brad Griesenauer, Hua Jiang, Djamilatou Adom, Pegah Mehrpouya-Bahrami, Srishti Chakravorty, Majid Kazemian, Tanbeena Imam, Rajneesh Srivastava, Tristan A. Hayes, Julian Pardo, Sarath Chandra Janga, Sophie Paczesny, Mark H. Kaplan, Matthew R. Olson
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Research Article Immunology Inflammation

Granzyme A–producing T helper cells are critical for acute graft-versus-host disease

Abstract

Acute graft-versus-host disease (aGVHD) can occur after hematopoietic cell transplant in patients undergoing treatment for hematological malignancies or inborn errors. Although CD4+ T helper (Th) cells play a major role in aGVHD, the mechanisms by which they contribute, particularly within the intestines, have remained elusive. We have identified a potentially novel subset of Th cells that accumulated in the intestines and produced the serine protease granzyme A (GrA). GrA+ Th cells were distinct from other Th lineages and exhibited a noncytolytic phenotype. In vitro, GrA+ Th cells differentiated in the presence of IL-4, IL-6, and IL-21 and were transcriptionally unique from cells cultured with either IL-4 or the IL-6/IL-21 combination alone. In vivo, both STAT3 and STAT6 were required for GrA+ Th cell differentiation and played roles in maintenance of the lineage identity. Importantly, GrA+ Th cells promoted aGVHD-associated morbidity and mortality and contributed to crypt destruction within intestines but were not required for the beneficial graft-versus-leukemia effect. Our data indicate that GrA+ Th cells represent a distinct Th subset and are critical mediators of aGVHD.

Authors

Sungtae Park, Brad Griesenauer, Hua Jiang, Djamilatou Adom, Pegah Mehrpouya-Bahrami, Srishti Chakravorty, Majid Kazemian, Tanbeena Imam, Rajneesh Srivastava, Tristan A. Hayes, Julian Pardo, Sarath Chandra Janga, Sophie Paczesny, Mark H. Kaplan, Matthew R. Olson

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Figure 3

IL-4 and IL-6 signaling synergize in the induction of GrA+ Th cells.

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IL-4 and IL-6 signaling synergize in the induction of GrA+ Th cells. (A)...
(A) Naive Th cells were differentiated in the presence of the indicated cytokines (TLR2 = PAM3CSK4) and stained for intracellular GrA. Data represents a single screen with cells pooled from 3 mice. These data are representative of 3 individual screening experiments. Nil, no treatment. (B) Indicated Th cells from 3 individual mice were cultured with/without IL-6 (10 ng/mL) followed by stimulation and intracellular staining for cytokines and GrA. Representative contour plots are depicted (left panel), and the frequency of GrA+ cells is depicted in the right panel. (C) Cells from 3 mice were cultured with IL-4, anti–IFN-γ, and increasing doses of IL-6. On day 5 of culture mRNA was measured by real-time PCR. Statistical analysis was done by Student’s t test, and reported values are corrected for multiple comparisons. (D) Cells from 3 individual mice were cultured with IL-4, IL-4+IL-6, or IL-4+IL-6 and with IL-21 (100 ng/mL) added at day 3 of culture. On day 5 of culture, cells were harvested and stained for intracellular GrA and GrB (left panel), and frequencies of GrA+ and GrB+ cells were determined by flow cytometry (right panel). Statistical analysis was done by 2-way ANOVA with Holm-Šidák correction for multiple comparisons. (E) Indicated Th subsets were analyzed by CyTOF and displayed in a dot overlays plot. Cytokine screening assays were performed 4 times with pooled naive CD4+ T cells. Th polarization experiments were repeated 3–6 times. Error bars represent standard deviation of the mean. CyTOF analysis was performed twice with pooled naive CD4+ T cells isolated from multiple animals.