Irf4 Is Correlated With The Conversion To A Th17-Like Phenotype In Regulatory T Cells From The Malignant Pleural Effusion

Background: ROR gamma t(+)Foxp3(+) (Th17-like) Tregs are a plastic Treg subset implicated in immune-related diseases; however, the mechanism of Treg phenotypic transformation in malignant pleural effusion (MPE) has not been elucidated.Methods: The percentage of CD4(+)CD25(+)Foxp3(+)Helios(+) and ROR gamma t(+)Foxp3(+) Tregs from peripheral blood and pleural effusion mononuclear cells were measured. The level of interferon regulatory factor 4 (IRF4) mRNA expression was detected by quantitative real-time reverse transcription polymerase chain reaction. The effects of IRF4 on the induction of Tregs from patients with non-small cell lung cancer (NSCLC) were evaluated in vitro. Correlation assays between IRF4 expression and the frequency of ROR gamma t(+)Foxp3(+) Tregs were performed.Results: The frequency of CD4(+)CD25(+)Foxp3(+)Helios(+) Tregs and CD4(+)ROR gamma t(+) Th17 cells was both increased in the MPE of NSCLC patients. The group of double-positive Foxp3(+)ROR gamma t(+) Treg phenotype were identified in the pleural effusion. A significant increase in the frequency of Foxp3(+)ROR gamma t(+) Tregs was found in MPE compared with the nonmalignant pleural effusion (NPE). Compared to NPE, the relative level of IRF4 expression was increased in the MPE. IRF4 expression was positively associated with the frequency of Foxp3(+)ROR gamma t(+) Tregs in the PE. In vitro, the level of Helios mRNA and protein expression was reduced in induced Tregs following IRF4 over-expression. Additionally, the level of ROR gamma t protein expression was substantially increased. However, ectopic Helios expression in induced Tregs reversed the effects induced by enhanced IRF4 expression.Conclusion: IRF4 may serve as a potential molecule that promotes the conversion of regulatory T cells from MPE to a Th17-like phenotype by modulating Helios.