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Gene expression profiling of NK cells isolated from the pancreas of BDC TCR expressing NOD mice before or after Treg ablation by DTR. Also, NK cells cultured in vitro with activating cytokines IL12 and IL-18 with or without regulatory TGFb.

ABSTRACT: Following Treg ablation in the BDC/NOD.Foxp3-DTR strain, NK cells produce IFNg and accumulate to higher percentage and number. We explored the signature pathways responsible for this phenomenon using microarray prolifing and comparison to other activation signatures.

ORGANISM(S): Mus musculus

PROVIDER: GSE39197 | GEO | 2013/06/20

SECONDARY ACCESSION(S): PRJNA170188

REPOSITORIES: GEO

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Gene expression profiling of NK cells isolated from the pancreas of BDC TCR expressing NOD mice before or after Treg ablation by DTR. Also, NK cells cultured in vitro with activating cytokines IL12 and IL-18 with or without regulatory TGFb.

Project description:Following Treg ablation in the BDC/NOD.Foxp3-DTR strain, NK cells produce IFNg and accumulate to higher percentage and number. We explored the signature pathways responsible for this phenomenon using microarray prolifing and comparison to other activation signatures. All microarray populations were obtained from highly purified (double sorted) NK cell populations from the pancreas or the spleen following Treg ablation. Triplicates were generated for all ex vivo data. Raw data were preprocessed with the RMA algorithm in GenePattern and averaged expression values were used for analysis.

2013-06-20 | E-GEOD-39197 | biostudies-arrayexpress

ScRNA-seq of stromal cells in WT and LRRC15 DTR mice

Project description:KPR tumors were subcutaneously implanted in Lrrc15DTRGFPwt/wt (DTR–) or Lrrc15DTRGFPwt/ki (DTR+) mice and DT treatment was initiated in both DTR– and DTR+ mice when tumors reached a mean volume of 100-200mm3. scRNAseq of CD24-CD45- stromal cells from a total of 40 tumor-bearing animals across both DTR– and DTR+ genotypes was carried out at four different time points including 10 days post tumor implantation and immediately prior to initiation of DT treatment (IOT) (referred to as day 0) and on days 7, 14, and 21 post IOT.

2022-08-17 | E-MTAB-12036 | biostudies-arrayexpress

Transcriptional profiling of NK cells sorted from lungs and axillary LNs from Cdh1F/F;Trp53F/F;Foxp3GFP-DTR mice bearing 100mm2 orthotopically transplanted mammary tumors, treated with difteria toxin (DT) or PBS [NK]

Project description:We studied the impact of DT treatment (Treg depletion) on NK cells (CD3-, NKp46+, DX5+) in axillary TDLNs and lungs of tumor bearing Foxp3GFP-DTR mice . Here we generated RNAseq data from sorted NK cells from DT or PBS treated Foxp3GFP-DTR mice bearing 100mm2 mammary tumors from lungs, axillary TDLNs

2022-03-01 | GSE169025 | GEO

Expression data from FITC challenged skin of CLEC4C-DTR mice vs. littermate controls treated with diptheria toxin

Project description:CLEC4C-DTR exhibit exacerbated contact hypersensitivty to the FITC hapten. Here we assess the transcriptional differences in whole skin of FITC challenged CLEC4C-DTR+ mice and DTR- mice treated with diptheria toxin. Overall, CLEC4C-DTR upregualated 439 genes and downregulated 152 genes compared to transgene negative controls, filtering output on 2 fold expression level difference and FDR<0.05.

2020-10-28 | GSE160261 | GEO

Expression profiles of satellite cells and alpha7Sca1 cells in mdxITGAM-DTR mice

Project description:We took advantage of a dystrophic mouse model of transient macrophage-depletion, mdxITGAM-DTR mice, in order to analyze the role of macrophage in skeletal muscle regeneration. We generated the transcriptome of satellite cells (SCs) and alpha7Sca1 cells purified by cell sorting from mdxITGAM-DTR mice. The mice were treated, by intramuscular injection, with PBS, as vehicle, or with Diphtheria toxin (DT) in order to achieve the macrophage depletion form hind-limb muscle We described a shift in identity of muscle stem cells dependent on the crosstalk between macrophages and satellite cells. Indeed macrophage depletion determines an exacerbated dystrophic phenotype associated with adipogenic conversion of SCs and reduction of the SC pool.

2019-09-17 | GSE134770 | GEO

FXTAS presents with upregulation of the cytokines IL12 and TNFα.

Project description:IntroductionFragile X Tremor and Ataxia Syndrome is a progressive neurodegenerative disorder that develops in some FMR1 premutation carriers. The objective of this study is to characterize how cytokine levels are altered in the FXTAS brain.MethodsFresh frozen cerebellar tissue from FXTAS cases and controls was homogenized and analyzed for 12 different cytokines using a commercially available ELISA panel.ResultsRelative to controls, FXTAS cases showed large and significant increases in the cytokines IL-12 and TNFα. There were large but non-significant increases in the levels of IL-2, IL-8, and IL-10 in FXTAS cases. The cytokines IL-1A, IL-1B, IL-4 IL-6, IL-17A, IFNγ, and GM-CSF were not different between FXTAS and control subjects.ConclusionsFor the first time, we demonstrate an increase in the pro-inflammatory cytokines TNFα and IL-12 in the FXTAS brain, both of which are implicated in the pathogenesis of Multiple Sclerosis, another neurodegenerative disorder that predominantly consists of white matter disease.

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Gene expression profiling of BDC2.5 CD4T cells isolated from NOD mice after in vivo antigen stimulation with either DEC205+ or DCIR2+ DCs.

Project description:We identified DCIR2+DCs but not DEC205+DCs as able to induce peripheral T cell tolerance in pre-diabetic autoimmune NOD mice. To determine what distinct genetic programs are elicited in the auto-reactive CD4 T cells early after stimulation by these two DC subsets, we utilized adoptive transfer of BDC2.5 CD4 T cells into NOD mice, which were then given chimeric antibody to deliver the beta-cell specific antigen to either DCIR2+DCs or DEC205+DCs, leading to BDC2.5 CD4 T cell specific stimulation in vivo. The analysis shows that the negative transcriptional factor Zbtb32 (ROG) is up-regulated more in BDC2.5 CD4 T cells after stimulated with a antigen via DCIR2+DCs presentation, compared with DEC205+DCs, suggesting the involvement of Zbtb32 in DCIR2+DCs-mediated auto-reactive T cell tolerance in disease ongoing NOD mice. The BDC2.5 CD4 T cells after 14 hour of injection with a 100 ng of anti-DCIR2-BDC, anti-DEC205-BDC antibody, or PBS as a control to NOD mice, were sorted by BD FACSAria and their expression profile was analyzed using Affymetrix chips. NOD mice with transferred BDC2.5 T cell were injected with 100 ng of anti-DCIR2-BDC (NOD_DCIR2-BDC), anti-DEC205-BDC antibody (NOD_DEC-BDC), or PBS (NOD_PBS) as a control. After 14 hours, BDC2.5 CD4 T cells were sorted by BD FACSAria and their expression profile was analyzed using Affymetrix microarray chips. Each sample has a biological duplicate. Raw data were preprocessed with the RMA algorithm in Partek, and averaged expression values were used for analysis.

2015-07-01 | E-GEOD-69554 | biostudies-arrayexpress

scRNA-seq analysis of dendritic cells in skin draining lymph nodes of CLEC4C-DTR mice treated with FITC hapten

Project description:Skin draining lymph node dendritic cell populations are known to be diverse and distinguishable via their migratory or resident properties as well as by lineage. We analyzed these populations at steady state and during the sensitization phase of the Th2-driven FITC-hapten contact hypersensitivity (CHS) model. We also analyzed the dendritic cells populations of CLEC4C-DTR+ mice treated with diptheria toxin, which demonstrate exacerbated pathology in the FITC-CHS model. We demonstrate the diversity of dendritic cell populations at steady state and during type 2 inflammation, and surprisingly, reveal a skewing of the DC populations in DT treated CLEC4C-DTR+ mice, with an enrichment of DC2 migratory populations and depletion of CD326+ migratory cells in addition to pDC.

2020-12-31 | GSE161604 | GEO

Synergy between Common ? Chain Family Cytokines and IL-18 Potentiates Innate and Adaptive Pathways of NK Cell Activation.

Project description:Studies to develop cell-based therapies for cancer and other diseases have consistently shown that purified human natural killer (NK) cells secrete cytokines and kill target cells after in vitro culture with high concentrations of cytokines. However, these assays poorly reflect the conditions that are likely to prevail in vivo in the early stages of an infection and have been carried out in a wide variety of experimental systems, which has led to contradictions within the literature. We have conducted a detailed kinetic and dose-response analysis of human NK cell responses to low concentrations of IL-12, IL-15, IL-18, IL-21, and IFN-?, alone and in combination, and their potential to synergize with IL-2. We find that very low concentrations of both innate and adaptive common ? chain cytokines synergize with equally low concentrations of IL-18 to drive rapid and potent NK cell CD25 and IFN-? expression; IL-18 and IL-2 reciprocally sustain CD25 and IL-18R? expression in a positive feedback loop; and IL-18 synergizes with Fc?RIII (CD16) signaling to augment antibody-dependent cellular cytotoxicity. These data indicate that NK cells can be rapidly activated by very low doses of innate cytokines and that the common ? chain cytokines have overlapping but distinct functions in combination with IL-18. Importantly, synergy between multiple signaling pathways leading to rapid NK cell activation at very low cytokine concentrations has been overlooked in prior studies focusing on single cytokines or simple combinations. Moreover, although the precise common ? chain cytokines available during primary and secondary infections may differ, their synergy with both IL-18 and antigen-antibody immune complexes underscores their contribution to NK cell activation during innate and adaptive responses. IL-18 signaling potentiates NK cell effector function during innate and adaptive immune responses by synergy with IL-2, IL-15, and IL-21 and immune complexes.

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IL12 immune therapy clinical trial review: Novel strategies for avoiding CRS-associated cytokines.

Project description:Interleukin 12 (IL-12) is a naturally occurring cytokine that plays a key role in inducing antitumor immune responses, including induction of antitumor immune memory. Currently, no IL-12-based therapeutic products have been approved for clinical application because of its toxicities. On the basis of this review of clinical trials using primarily wild-type IL-12 and different delivery methods, we conclude that the safe utilization of IL-12 is highly dependent on the tumor-specific localization of IL-12 post administration. In this regard, we have developed a cell membrane-anchored and tumor-targeted IL-12-T (attIL12-T) cell product for avoiding toxicity from both IL-12 and T cells-induced cytokine release syndrome in peripheral tissues. A phase I trial using this product which seeks to avoid systemic toxicity and boost antitumor efficacy is on the horizon. Of note, this product also boosts the impact of CAR-T or TCR-T cell efficacy against solid tumors, providing an alternative approach to utilize CAR-T to overcome tumor resistance.

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