CD4 T cell expression of PTIP is necessary for epigenetic maintenance of Th1/Th17 responses during Cryptococcus neoformans infection

Abstract

Abstract Containment and clearance of opportunistic fungal pathogen Cryptococcus neoformans (CN) requires Th1/Th17 polarized CD4 T cells. Studies show that Th1/Th17 T-cell polarization is linked to enriched chromatin activation at histone 3 lysine 4 (H3K4). DNA repair enzyme PTIP forms a complex with MLL3/4 H3K4 methyl transferases, but its role in regulation of immune-related genes remains unknown. We hypothesized that the absence of PTIP in CD4 T cells would result in a defect in generation and/or maintenance of Th1/Th17 driven immune protection against CN infection. CD4Cre+, PTIP knockout mice (CD4-PTIP-KO) and their control Cre- littermates were infected with CN and their fungal burden and T cell polarization assessed at 3 and 6 weeks post-infection (wpi). There were few statistically significant differences at 3 wpi, but no defects in IFNγ and TNFαproduction in these mice. However, by 6 wpi we observed profound defects in fungal clearance and the expression of hallmarks of Th1 and Th17 responses. CD4-PTIP-KO mice had diminished fungal clearance by 1.5-log and showed diminished frequency of IFNγ, TNFα, and IL-17 producing CD4 T-cells as well as suppressed global production of these cytokines. Furthermore, frequency of IFNγ-IL-17 dual-producing CD4 T cells, previously found to correlate with clearance, was diminished in CD4-PTIP-KO mice. Interestingly no T-cell activation defect (frequency of CD44hi and CD62L low) was altered in CD4-PTIP-KO mice. Collectively, we conclude that PTIP is not necessary for induction of Th1 and Th17 polarized T cells but it is crucial for maintaining robust Th1/Th17 cytokine production by polarized T cells throughout C.neo infection, presumably via epigenetic stabilization of CD4 T cells Th1/Th17 programing.