Comparison of immature BMDC to BMDC after nkt cell modulation
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE18547 | GEO | 2009/10/17
SECONDARY ACCESSION(S): PRJNA121357
REPOSITORIES: GEO
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Project description:Analyze the gene expression profile of BMDC before and after nkt cell modulation. Total RNA was extracted from CD11c cells with TRIzol Reagent, purified with RNeasy Kit (Qiagen) and subsequently amplificated using the one cycle target labeling kit according to the AffymetrixGeneChip® Expression Analysis Technical Manual (Affimetrix). Fragmented cRNAs were probed to AffymetrixGeneChip® Mouse Genome 430 2.0 arrays for 16 hours at 45°C in the AffymetrixHybridization Oven 640. The arrays were then washed and stained with streptavidin-phycoerythrin using the AffymetrixFluidic Station 450/250 and scanned using the AffymetrixGeneChip® Scanner 3000. The images were analyzed using GCOC® Software and comparison analyses carried out according to the instructions provided by Affymetrix. Each experiment was performed in triplicates and repeated two times. Data handling was mainly done using Bioconductor Software. The probe set-based summary data were log transformed and normalized for probe set intensity-dependent biases. The identification of differentially expressed genes was addressed using linear modeling approach and empirical Bayer methods together with false discovery rate correction of the p-value. We considered a gene to be differentially increased in nkt-DCs only if its expression intensity was significantly changed (p<0.001) compared to Untr-DCs.
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Project description:Comparison of immature BMDC to BMDC after nkt cell modulation
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Project description:We report the identification of immature thymic CD4(-),CD8(-) double-negative (DN)1e cells with the NKT cell lineage potential. We also analyzed the gene expression profiles of DN1e thymocytes compared with those of mature thymic NKT cell developmental stages termed NKT stage-1, 2, and -3, which are characterized by differential expression levels of NK1.1 and CD44 antigens in C57BL/6J mouse strain.
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Project description:Semi-invariant natural killer T (NKT) cells are thymus-derived innate lymphocytes that modulate microbial and tumour immunity as well as autoimmune diseases. These immunoregulatory properties of NKT cells are acquired during their development. Much has been learnt regarding the molecular and cellular cues that promote NKT cell development, yet how these cells are maintained in the thymus and the periphery and how they acquire functional competence are incompletely understood. We found that IL-15 induced several Bcl-2 family survival factors in thymic and splenic NKT cells in vitro. Yet, IL15-mediated thymic and peripheral NKT cell survival critically depended on Bcl-xL expression. Additionally, IL-15 regulated thymic developmental stage 2 (ST2) to ST3 lineage progression and terminal NKT cell differentiation. Global gene expression analyses and validation revealed that IL-15 regulated Tbx21 (T-bet) expression in thymic ST3 NKT cells. The loss of IL15-dependent T-bet expression resulted in poor expression of IFN-γ and several NK cell receptors in NKT cells. Taken together, our findings reveal a critical role for IL-15 in NKT cell survival, which is mediated by Bcl-xL, and effector differentiation, which is regulated by T-bet. Gene expression was measured in NKT cells sorted from pooled thymi of wild-type (3 replicates) or IL-15 deficient (2 replicates) mice.
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Project description:Semi-invariant natural killer T (NKT) cells are thymus-derived innate lymphocytes that modulate microbial and tumour immunity as well as autoimmune diseases. These immunoregulatory properties of NKT cells are acquired during their development. Much has been learnt regarding the molecular and cellular cues that promote NKT cell development, yet how these cells are maintained in the thymus and the periphery and how they acquire functional competence are incompletely understood. We found that IL-15 induced several Bcl-2 family survival factors in thymic and splenic NKT cells in vitro. Yet, IL15-mediated thymic and peripheral NKT cell survival critically depended on Bcl-xL expression. Additionally, IL-15 regulated thymic developmental stage 2 (ST2) to ST3 lineage progression and terminal NKT cell differentiation. Global gene expression analyses and validation revealed that IL-15 regulated Tbx21 (T-bet) expression in thymic ST3 NKT cells. The loss of IL15-dependent T-bet expression resulted in poor expression of IFN-γ and several NK cell receptors in NKT cells. Taken together, our findings reveal a critical role for IL-15 in NKT cell survival, which is mediated by Bcl-xL, and effector differentiation, which is regulated by T-bet.
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