Human TH9 differentiation is dependent on signal transducer and activator of transcription (STAT) 3 to restrain STAT1-mediated inhibition

Abstract

Background: Patients with loss-of-function (LOF) signal transducer and activator of transcription 3 (STAT3) mutations have dermatitis, enhanced IgE production despite a relative lack of immediate hypersensitivity, recurrent infection, and an increased rate of lymphoma in addition to a number of skeletal and connective tissue abnormalities. Patients with STAT1 gain-of-function (GOF) mutations also have susceptibility to candidiasis and sinopulmonary infection, as well as autoimmunity and squamous cell carcinoma, in addition to even more broad phenotypes.

Objective: Because of the link between T_H9 cells and allergic inflammation, autoimmunity, and antitumor surveillance and because evidence shows a role for either STAT3 or STAT1 in T_H9 differentiation conflicts, we sought to determine the status on this lineage of STAT1 GOF and STAT3 LOF mutations in human subjects.

Methods: We detected IL-9 levels and T_H9 differentiation in patients with STAT3 LOF and STAT1 GOF mutations, together with T_H9 transcript factors, and partially rescued their deficiency in vitro by adding cytokines they lacked or transfecting key molecules.

Results: We found that PBMCs or sorted naive CD4⁺ T cells from patients with STAT3 LOF and STAT1 GOF mutations had impaired T_H9 generation/differentiation. STAT3 inhibition in normal T_H9 cultures diminished early IL-21 induction and late IL-9 production, whereas exogenous IL-21 enhanced T_H9 differentiation, even with STAT3 inhibition, by restoring suppressor of cytokine signaling 3 expression and thus inhibiting excessive phosphorylated signal transducer and activator of transcription (p-STAT) 1 activation. Furthermore, exogenous expression of suppressor of cytokine signaling 3 or either T-bet or STAT1 RNA interference in STAT3 LOF cells partially rescued IL-9 differentiation.

Conclusion: Collectively, these results suggest that human T_H9 differentiation depends on normal p-STAT3 and IL-21 production to suppress p-STAT1 activation and T-bet transcription.

Keywords: Human T(H)9; IL-21; allergy; anticancer; signal transducer and activator of transcription 1; signal transducer and activator of transcription 3.