Prier, J. E. | bioRxiv Science

bioRxiv2020

KDM6B-dependent chromatin remodelling underpins effective virus-specific CD8+ T cell differentiation.

Naive CD8+ T cell activation results in an autonomous program of cellular proliferation and differentiation. However, the mechanisms that underpin this process are unclear. Here we profiled genome-wide changes in chromatin accessibility, gene transcription and the deposition of a key chromatin modification (H3K27me3) early after naive CD8+ T cell activation. Rapid upregulation of the histone demethylase, KDM6B, prior to first cell division was required for initiating H3K27me3 removal at genes essential for subsequent T cell differentiation and proliferation. Inhibition of KDM6B-dependent H3K27me3 demethylation limited the magnitude of an effective primary virus-specific CD8+ T cell response and the formation of memory CD8+ T cell populations. Accordingly, we define the early spatio-temporal events underpinning early lineage-specific epigenetic reprogramming that is necessary for autonomous CD8+ T cell proliferation and differentiation.

immunology↗

bioRxiv2019

Development of a KDM6b shRNA conditional knock down mouse model

Antigen-specific CD8+ T cell differentiation in response to infection is associated with specific changes in the chromatin landscape resulting in acquisition of the lineage-specific effector functions required for pathogen clearance. Lysine (K)-specific demethylase 6B (KDM6b) is a histone demethylase that specifically recognizes and removes methyl groups from K27 tri/dimethylation on histone 3. This histone modification is associated with a repressive transcriptional state, or, in combination with the active H3K4me3 mark, a bivalent epigenetic state. Resolution of bivalency at fundamental transcription factor loci has been shown to be a key mechanism for the initiation of CD8+ T cell differentiation. To begin to address the role of KDM6b in regulating H3K27me3 demethylation in CD8+ T cell responses to infection, a model whereby KDM6b levels can be modulated is needed. To address this, we developed a conditional short hairpin RNA (shRNA) mouse model targeting KDM6b. Here we demonstrate that KDM6b knockdown results in diminished naive, CD4+ and virus-specific CD4+ and CD8+ T cell response in response to influenza A infection. To address the molecular mechanism, we demonstrate that KDM6b knockdown resulted in reduced H3K27me3 removal from the Tbx21 bivalent promoter, compared to luciferase hairpin controls. Surprisingly, this did not necessarily impact T-BET expression, or resolution of other bivalent transcription factor promoters. These data suggest that KDM6b knockdown resulting in diminished IAV-specific CD8+ T cell responses may reflect a demethylase independent function.

immunology↗