ABSTRACT
Group 3 innate lymphoid cells (ILC3) have a prominent role in the maintenance of intestine mucosa homeostasis. The hypoxia-inducible factor (HIF) is an important modulator of immune cell activation and a key mechanism for cellular adaptation to oxygen deprivation. However, its role on ILC3 is not well known. In this study, we investigated how a hypoxic environment modulates ILC3 response and the subsequent participation of HIF-1 signaling in this process. We found increased proliferation and activation of intestinal ILC3 at low oxygen levels, a response that was phenocopied when HIF-1α was chemically stabilized and was reversed when HIF-1 was blocked. The increased activation of ILC3 relied on a HIF-1α-dependent transcriptional program, but not on mTOR-signaling or a switch to glycolysis. HIF-1α deficiency in RORyt compartment resulted in impaired IL-17 and IL-22 production by ILC3 in vivo, which reflected in a lower expression of their target genes in the intestinal epithelium and an increased susceptibility to Clostridiodes difficile infection. Taken together, our results show that HIF-1α activation in intestinal ILC3 is relevant for their functions in steady state and infectious conditions.
Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Hypoxia/immunology , Hypoxia/metabolism , Immunity, Innate , Lymphocyte Subsets/immunology , Lymphocyte Subsets/metabolism , Animals , Clostridium Infections/etiology , Clostridium Infections/metabolism , Disease Models, Animal , Disease Susceptibility , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Lymphocyte Activation/genetics , Lymphocyte Activation/immunology , Mice , Mice, Knockout , Mitochondria/metabolism , Nuclear Receptor Subfamily 1, Group F, Member 3/metabolism , Protein Stability , Signal Transduction , TOR Serine-Threonine Kinases/metabolismABSTRACT
INTRODUCTION: Periodontitis is characterized by inflammatory mediators beyond T lymphocyte function and phenotype (Th1/Th2/Th17). The clinical diversity in periodontitis makes it difficult to characterize the immune response in patients. This study evaluated the profile of the adaptive immune response in the periodontal disease model. METHODS: 72 rats (Wistar) were divided into a control group (CTL/day 0) and periodontitis (PD15/15 days and PD60/60 days). In the PD15 and PD60 groups, periodontal disease was induced by ligature with a silk thread placed in the cervical region of the upper first molar. After euthanasia, the periodontal tissue was analyzed by flow cytometry (CD4, CD8, CD25, CD44), semi-quantitative RT-PCR (T-bet, GATA-3, RORγt), semi-quantitative RT-PCR and ELISA IFN-γ, TNF-α, IFN-γ, IL-4, IL-6, IL-10, IL-17) and by Western blotting (Caspase-9, PCNA). RESULTS: The number of CD4+CD25+, CD4+CD44+, CD8+CD25+ and CD8+CD44+ cells and expression levels of T-bet and GATA-3 are increased in the PD60 group compared to PD15 and CTL. The RORγ-t gene transcript increased in the PD15 group in relation to PD60 and CTL. The cytokines IFN-γ, TNF-α and IL-17 increased in the PD60 group in relation to PD15. The expression of Caspase-9 was higher in the PD60 group than in PD15. CONCLUSIONS: The results suggest that the evolution of gingivitis to periodontitis is related to the accumulation of activated Th1 cells (IFN-γ and TNF-α) associated with the presence of increased IL-17. Studies with inhibitors of these cytokines in periodontal disease may lead to therapy directed at blocking the inflammatory process in this pathology, interrupting bone loss.
