Epigenetic Regulation of Immune Dysfunction in Chronic Prostatitis/Chronic Pelvic Pain Syndrome (CP/CPPS)

Background: Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a prevalent and debilitating condition with unclear etiology. Increasing evidence implicates immune dysregulation, yet the molecular mechanisms underlying impaired immune regulation remain poorly defined. This study investigated the role of adaptive immune responses and DNA methylation in CP/CPPS pathogenesis. Methods: Voided bladder 3 (VB3) urine samples from CP/CPPS patients and healthy controls were analyzed for CD4⁺ T cell markers and lineage-defining transcription factors. DNA methylation profiling of peripheral blood mononuclear cells (PBMCs) and purified CD4⁺ T cells was performed using targeted methylation arrays. Functional assays evaluated IL-10 production following lipopolysaccharide (LPS) stimulation, with or without azacitidine (AZA), a DNA methyltransferase inhibitor that reverses methylation-dependent gene silencing. In vivo relevance was assessed using the experimental autoimmune prostatitis (EAP) mouse model. Results: VB3 samples from CP/CPPS patients demonstrated elevated CD4⁺ T cell transcripts with a Th17/Th1 (RORγT⁺/TBET⁺) bias and reduced FOXP3 expression. DNA methylation analysis revealed hypermethylation of IL10 and FOXP3 promoters and hypomethylation of inflammatory genes including CD274, ITGAL, and TNFα. PBMCs from patients exhibited diminished IL-10 secretion in response to LPS, which was restored by azacitidine treatment. In the EAP model, recombinant IL-10 administration failed to attenuate pelvic allodynia, whereas azacitidine significantly reduced pain sensitivity. Conclusions: CP/CPPS is associated with widespread epigenetic reprogramming of immune regulatory genes leading to impaired IL-10-mediated immune regulation. Pharmacologic inhibition of DNA methylation restored IL-10 responses and alleviated pain in vivo, supporting demethylation therapy as a potential strategy for treating chronic prostatic inflammation and pelvic pain.