Project description:Purpose: To characterise gene expression of in vitro expanded mouse gammadelta T cells. Methods: Freshly isolated and IL-2 expanded gammadelta T cells, single samples, and total RNA was isolated and sequenced. Results: High upregulation of Th1 like genes including several granzymes genes. Conclusion: Expanded gammadelta T cells express Th1-like genes.
Project description:This SuperSeries is composed of the following subset Series: GSE31255: Isolation and in vitro expansion of human colonic stem cells [Expression profile] GSE31256: Isolation and in vitro expansion of human colonic stem cells [CGH] Refer to individual Series
Project description:Psoriasis is a chronic skin suffering with multiple comorbidities such as psoriatic arthritis and cardiovascular diseases. Increasing evidences have shown the γδ T cells as the sources of IL-17A play critical roles in the psoriatic syndrome. However, there is still lack an effective way to manipulate these pathogenic γδ T cells which were less studied relative to ab T cells. The present study aims to characterize the phenotype of γδ T cells and evaluate the impact of D-mannose (a C-2 epimer of glucose) on γδ T-mediated psoriasis. We found the psoriatic γδ T cells underwent robust proliferation and acquired an IL-17 producing phenotype. The transcriptomic profiles of these skin draining LN γδ T cells had elevated glycolytic features. Importantly, Treatment of D-mannose inhibited γδ T cells and successfully alleviated the local and systematic inflammations induced by imiquimod. The decreased AKT/mTOR/HIF-1a signaling and glycolytic ability may contribute to the suppression of γδ T cells achieved by mannose. Our study deepened the understandings of γδ T cells in psoriasis, meanwhile, promoted D-mannose utilization as a potent clinical application for γδ T cells driven autoimmune diseases.
Project description:The mammalian gastrointestinal tract harbors thousands of bacterial species that include symbionts as well as potential pathogens. The immune responses that limit access of these bacteria to underlying tissue remain poorly defined. In this study, we used microarrays to uncover the transcriptional responses that occur in small intestinal γδ intraepithelial lymphocytes following bacterial challenge. γδ intraepithelial lymphocytes (γδ IEL) were isolated by flow cytometry from the small intestines of germ-free mice, or from age- and sex-matched conventionally-raised counterparts. We extracted RNAs from these purified γδ IEL for analysis on Affymetrix DNA microarrays. The mice were all >8 weeks in age, and each sample represents a pool of RNAs from 5-8 mice.
Project description:Here, we investigated γδ T cells in a longitudinal cohort of preterm neonates during the first year of life. Our data reveal dynamic postnatal maturation patterns of γδ T cell subsets, which are largely age-dependent. We report on the expansion of fetal-derived γδ T cells in preterm neonates with sepsis. Single cell transcriptome analyses identified HLA-DRhigh and CD83+ γδ T cells in neonatal sepsis.
Project description:Because activation of the immune response is dependent on extensive changes in gene expression, it is likely that a major component of inter-individual variation in the immune response is ultimately mediated at the level of gene regulation. Here, we examine the influence of genetic variation on inducible gene expression in the murine immune response. We extracted primary CD4 splenocytes from inbred strains A/J, C57BL/6J, BALB/c, DBA/2J, and 129x1/SvJ (3 animals/strain) and measured mRNA transcript levels using microarrays in both basal state and four hours after stimulation with PMA/Ionomycin. We report strain-specific genome-wide differences in mRNA expression between A/J, Balb/c, C57BL/6J, DBA/2J and 129X1/SvJ inbred mice in CD4 splenocytes in basal, stimulated (4hr, PMA/I) states.
Project description:During the course of infection, T cells are confronted with a multitude of non self epitopes but only respond to a few epitopes and neglect other potentially immunogenic peptides. Restriction of T cell responses to a small number of selected epitopes (immunodominance) is a central feature of immune responses. Immunodominance is considered to be disadvantageous to the host because it allows pathogens to escape by selectively mutating the relevant epitope(s). Using Affymetrix microarrays, we compared the gene expression profile of unprimed CD8 T cells to that of effector CD8 T cells specific for a dominant (H7a) and a nondominant (HY) Ag. Our key finding is that T cells specific for the dominant and nondominant Ags displayed similar gene expression profiles except for a few gene transcripts, such as Gzma, Sell, Il7r and Klrg1, that contribute to the fitness of effector CD8 T cells. The differences between HY- and H7a-specific CD8 T cells were validated by real-time PCR and flow cytometry analyses. We propose that, by leading to selective expansion of the fittest CD8 effector T cells, immunodominance may be beneficial to the host. Inhibition of T cell response to nondominant Ags would ensure that host resources (APCs, cytokines) for which T cells compete are devoted to T cells that have the best effector potential. One implication is that, in general, favouring expansion of the fittest effector T cells may be more important that increasing the diversity of the T cell repertoire. Experiment Overall Design: B10.H7b female mice were primed by i.p. injection of a cell mixture containing 2 x 107 B10 male splenocytes and 2 x 107 B10.H7b male splenocytes. On day 14 after priming, splenocytes were labeled with anti-CD8 Ab as well as H7a- and HY-Tet labeled with different fluorochromes. Then, three populations of splenocytes were purified using a FACS cell sorting: HYTet+ (n=4), H7aTet+ (n=4), and Tet- (n=5) CD8 T cells. RNA of sorted T cells was extracted and linearly amplified, cRNA was prepared, and Affymetrix Mouse Genome 430 2.0 oligonucleotide arrays were used to analyze gene expression.
Project description:Triple negative breast cancer (TNBC) lacks targeted therapy options. TNBC is enriched in breast cancer stem cells (BCSCs), which play a key role in metastasis, chemoresistance, relapse and mortality. γδ T cells hold great potential in immunotherapy against cancer, and might be an alternative to target TNBC. γδ T cells are commonly observed to infiltrate solid tumors and have an extensive repertoire of tumor sensing, recognizing stress-induced molecules and phosphoantigens (pAgs) on transformed cells. We show that patient derived triple negative BCSCs are efficiently recognized and killed by ex vivo expanded γδ T cells from healthy donors. Orthotopically xenografted BCSCs, however, were refractory to γδ T cell immunotherapy. Mechanistically, we unraveled concerted differentiation and immune escape: xenografted BCSCs lost stemness, expression of γδ T cell ligands, adhesion molecules and pAgs, thereby evading immune recognition by γδ T cells. Indeed, neither pro-migratory engineered γδ T cells, nor anti-PD 1 checkpoint blockade significantly prolonged overall survival of tumor-bearing mice. BCSC immune escape was independent of the immune pressure exerted by the γδ T cells, and could be pharmacologically reverted by Zoledronate or IFN-α treatment. These results pave the way for novel combinatorial immunotherapies for TNBC.