Project description:We have conducted single cell-RNA, -TCR, -ATAC sequencing (10x Genomics) to investigate a difference of gene expression, TCR repertoir and chromatin accesibility of thymic iNKT cells from wild-type and CD4-Cre-mediated T cell-specific Prkd2/3 doubly deficirnt (Prkd2/3ΔCD4) mice. Results demonstrate that Prkd2/3 are required for iNKT cell development and formaion of proper iNKT cell subtypes.
Project description:We have conducted TCR a/b profiling to compare the usage of TCR a/b chains of thymic iNKT cells from wild-type (WT) and CD4-Cre-mediated T cell-specific Prkd2/3 doubly deficirnt (Prkd2/3ΔCD4) mice. Results demonstrate that top one frequent TCRa chain is identical, whereas usage of TCRb chain is different between WT and Prkd2/3ΔCD4 iNKT cells.
Project description:Single-cell TCR-seq analysis of murine thymic iNKT cells from three independent BALB/c mice and three independent Cd80/Cd86 (B7)-deficient mice
Project description:iNKT cells show significant lineage diversity among mouse strains. We used scRNA-seq to analyze the diversity of thymic iNKT cells between BALB/c and B6 mice.
Project description:iNKT cells show significant lineage diversity among mouse strains. We used scTCR-seq to analyze the diversity of thymic iNKT cells between BALB/c and B6 mice.
Project description:Development of T cells is controlled by the signal strength of the TCR. The scaffold protein Kinase D-interacting substrate of 220 kDa (Kidins220) binds to the TCR; however, its role in T cell development was unknown. Here, we show that T cell-specific Kidins220 knock-out (T-KO) mice have strongly reduced invariant natural killer T (iNKT) cell numbers and modest decreases in conventional T cells. Enhanced apoptosis due to increased TCR signaling in T-KO iNKT thymocytes of developmental stage 2 and 3 shows that Kidins220 downregulates TCR signaling at these stages. scRNAseq indicated that the transcription factor Aiolos is downregulated in Kidins220-deficient iNKT cells. Analysis of an Aiolos KO demonstrated that Aiolos is a downstream effector of Kidins220 during iNKT cell development. In the periphery, T-KO iNKT cells show reduced TCR signaling upon stimulation with α-galactosylceramide, suggesting that Kidins220 promotes TCR signaling in peripheral iNKT cells. Thus, Kidins220 reduces or promotes signaling dependent on the iNKT cell developmental stage.