Project description:gd T cell infiltration into tumours usually correlates with improved patient outcome, but both tumour-promoting and tumoricidal effects of γδ T cells have been documented. Human γδ T cells can be divided into functionally distinct subsets based on T cell receptor Vd usage. Still, the contribution of these different subsets to tumour immunity remains elusive. Here, we provide a detailed gd T cell profiling in colon tumours, comprising mRNA quantification using the Nanostring platform, in combination with mass and flow cytometry and TCR sequencing. δ chain usage in both the macroscopically unaffected colon mucosa and tumours varied considerably between patients, with substantial fractions of Vδ1, Vδ2, and non-Vd1Vd2 cells. Nanostring analyses of flow cytometry sorted Vd1, Vd2 and non-Vd1Vd2 cells showed a large variation in gd T cell subsets between individual tumours, and we suggest that individual gd T cell composition in colon tumours may contribute to the balance between favourable and adverse immune responses, and thereby also patient outcome.
Project description:We compared gammadelta T-cell development and subsets in the thymus of mice in which Shh had been conditionally deleted from thymic epithelial cells using FoxN1-Cre to WT mice, and to transgenic mice in which a constitutively activator (Gli2DN2) or constitutuvely repressor (Gli2DC2) form of Gli2 were expressed under the control of the lck-promotor. We sorted CD27+CD3+TCR-gammadelta+ cells from 4 week old mice and comapred gene expression by RNAsequencing. All mice were on a C57BL/6 background.
Project description:Gammadelta T cells from Ugandan children were tested for their responsiveness to stimulation with malaria parasites. Independent PBMC samples from children were incubated with Plasmodium falciparum-infected red blood cells for 6 h post-treatment, and Vdelta2+ gammadelta T cells were double sorted by FACS directly into an RNA lysis buffer. RNA was isolated, amplified using 2 rounds of linear amplification, and hybridized to Agilent Human 8x60k microarrays as a single color experiment.
Project description:Natural killer (NKT) T cells exhibit tissue distribution, surface phenotype, and functional responses that are strikingly different from those of conventional T cells. The transcription factor PLZF is responsible for most of these properties, as its ectopic expression in conventional T cells is sufficient to confer to them an NKT-like phenotype. The molecular program downstream of PLZF, however, is largely unexplored. Here we report that PLZF regulates the expression of a surprisingly small set of genes, many with known immune functions. This includes several established components of the NKT cell developmental program. Transcriptional program downstream of PLZF in gammadelta NKT cells was analyzed by comparing wt, heterozygous and PLZF-deficient gammadelta NKT cells