The effects of prostaglandin E2 on the gene expression of human natural killer cells
Ontology highlight
ABSTRACT: Tumour immune escape is a major factor contributing to cancer progression and unresponsiveness to cancer therapies. Tumours can produce prostaglandin E2 (PGE2), an inflammatory mediator that directly acts on NK cells to inhibit anti-tumour immunity. However, it is unclear precisely how PGE2 influences NK cell tumour-restraining functions. Here, we investigated how treatment with PGE2 over 24 hours affects gene expression in human NK cells.
Project description:Oncolytic viruses (OV) kill tumour cells directly and induce anti-tumour immunity. We analysed gene expression profiles in human NK cells following treatment of PBMC with oncolytic reovirus in vitro.
Project description:STAT1 is an important regulator of NK cell maturation and cytotoxicity. Although the consequences of Stat1-deficiency have been described in detail the underlying molecular functions of STAT1 in NK cells are only partially understood. Here we describe a novel non-canonical role of STAT1 that was unmasked in NK cells expressing Stat1-Y701F. This mutation prevents JAK-dependent phosphorylation, subsequent nuclear translocation and cytokine-induced transcriptional activity. As expected Stat1-Y701F mice displayed impaired NK cell maturation comparable to Stat1-/- animals. In contrast Stat1-Y701F NK cells exerted a significantly enhanced cytotoxicity in vitro and in vivo suggesting a so-far unknown cytoplasmic function. Using immunofluorescence technology we uncovered the recruitment of STAT1 to the immunological synapse during NK cell killing. A Stat1ind mouse expressing FLAG-tagged STAT1α was used to study the STAT1α interactome in NK cells. Mass spectrometry revealed that STAT1 directly binds proteins involved in cell junction formation and proteins associated to membrane or membrane-bound vesicles. We propose a novel function for STAT1 in the immunological synapse of NK cells regulating tumor surveillance and cytotoxicity.
Project description:NK cells are part of the innate immune system and therefore directly involved in the response to a Candida albicans infection. Our aim is to better understand the interaction of NK cells and C.albicans cells. Natural Killer cells (NK cells) were isolated from blood of 3 different donors and coincubated with Candida albicans. Transcriptome analysis was performed after 3 and 6 hours using an Illumina HumanHT-12 v4 Expression BeadChip.
Project description:is involved in the formation of immune signaling complexes. To date only limited and moreover 35 conflicting data exist regarding the role of ADAP in NK cells. To extend existing knowledge we 36 investigated ADAP-dependency of NK cells in the context of in vivo infection with the intracellular 37 pathogen Listeria monocytogenes (Lm). Ex vivo analysis of infection-primed NK cells revealed 38 impaired expression of IFN-γ and chemokines as well as impaired cytotoxic capacity in NK cells 39 lacking ADAP as indicated by reduced CD107a surface expression and inefficient perforin 40 production. However, ADAP-deficiency had no global effect on NK cell morphology or intracellular 41 distribution of CD107a-containing vesicles. Proteomic definition of ADAPko and wild type NK cells 42 did not uncover obvious differences in protein composition during the steady state and moreover, 43 similar early response patterns were induced in NK cells upon infection independent of the genotype. 44 In line with protein network analyses that suggested an altered migration phenotype in naïve 45 ADAPko NK cells, in vitro migration assays uncovered significantly reduced migration of both naïve 46 as well as infection-primed ADAPko NK cells compared to wild type NK cells. We propose that this 47 migration defect might account at least in part for the fact that during in vivo infection significantly 48 lower numbers of ADAPko NK cells accumulate in the spleen i.e. the site of infection. In conclusion, 49 we show here that during systemic Lm infection in mice ADAP is essential for efficient cytokine and 50 chemokine production, cytotoxic capacity and migration of NK cells.
Project description:Natural killer (NK) cells contribute to immunosurveillance and first-line defense in the control of tumor growth and metastasis diffusion. NKEVs are constitutively secreted, are biologically active, reflect the protein and genetic repertoire of their originating cells and exert anti-tumor activity in vitro and in vivo. NKEVs from tumor-conditioned NK cells interact with naïve NK cells promoting their cytotoxic activity. In cancer NK cells exhibit profound defects in degranulation ability, a status probably reflected by their NKEVs. Hence, NKEVs could contribute to improve cancer therapy by interacting with tumor and/or immune cells at the same time sensing the actual NK cell status in cancer patients. Here we investigated the role of NKEVs in stimulating the immune system and developed an immune enzymatic test (NKExoELISA) to sense the systemic NK cell status by measuring plasma NK-derived exosomes through combined capture of exosomes, expressing typical EV (tsg101) and NK cell (CD56) markers. We analyzed by LC-MS/MS the protein content from NKEVs evaluating proteins differentially expressed in exosomes (NKExo), vescicles (NKMV) and total cell extract (Tot extr) from parental NK cells. Proteomic data confirmed the presence of many EV markers and detected several proteins involved in immune response, cell adhesion and complement biological processes.
Project description:Obesity is linked to an increased risk of many cancers and can impair the anti-tumour immune response. This project examined the effect of different sources of dietary fats on tumour immunity using a syngeneic model of melanoma in mice fed high fat diets (HFD) derived from different sources. HFD based on beef tallow, lard and butter increased tumour growth in this model, while HFD derived from coconut oil, palm oil or olive oil did not. Further experiments focusing on butter and palm oil based HFD found differential regulation of natural killer (NK) cells and CD8 T-cell infiltration and function in the tumour microenvironment, and identified enrichment in immunosuppressive long chain acylcarnitine species in mice fed a HFD based on butter compared to palm oil. This dataset contains proteomic data from NK cells from mice fed a standard fat diet (SFD), NK cells from mice fed a HFD based on butter and from mice fed a HFD based on palm oil.
Project description:We analyzed gene expression profiles of IL-18 generated murine NK cells in comparison to unstimulated, freshly isolated splenic NK cells. We identified a set of 1414 Affymetrix probe sets showing significant misregulation (Welch's T-test, p<0.05; Benjamini-Hochberg FDR corrected). IL-18 generated as well as unstimulated NK cells were isolated in three independent preparations and used for RNA extraction and hybridization on Affymetrix microarrays.
Project description:In order to identify an expression signature that might characterize the impact of Plasmodium parasites on NK cells, we have used Affymetrix oligonucleotide microarrays to examin the global gene expression profile of primary NK cells that have been co-incubated with P. falciparum infected red blood cells and have compared it to the expression pattern of NK genes induced by cytokine treatment. PBMCs were isolated from 3 healthy donors and were incubated with iRBCs, uRBCs, IL12 plus IL18 and without any stimulus. After 24 hours at 37ºC and 5%CO2, Natural killer cells were isolated by negative selection, checked for purity by flow cytometry and processed for RNA extraction and analysis with the Human Gene 1.0 Affymetrix array. The experiment was repeated 3 times (1-2 weeks apart) for each one of the 3 donors.
Project description:Two major subsets of rat natural killer (NK) cells can be distinguished based on their expression of either the Ly49s3 or the NKR-P1B lectin-like receptor. Ly49s3+ NK cells, but not NKR-P1B+ NK cells, express a wide range of Ly49 receptors. Here, we performed a global gene expression profiling of sorted, single-positive NKR-P1B+ or Ly49s3+ splenic NK cells in order to get an overview of differences in expressed genes between the NKR-P1B+ and Ly49s3+ NK cell subsets. NKR-P1B or Ly49s3 single-positive NK cells were sorted by flow cytometry from NK-enriched mononuclear cells from rat spleens. 9 rats of the PVG.7b strain aged 8-12 weeks were used. Total RNA was harvested by Tri Reagent extraction, and subjected to microarray analysis (Affymetrix Rat Genome 230 2.0 Array).
Project description:NK cells may acquire under certain conditions features of adaptive immune cells. As the functional role of memory NK cells in cancer has so far remained elusive, we reasoned whether tumor-priming itself might promote memory NK cell generation. We provide substantial evidence that independent from pro-inflammatory stimulation, tumor-induced memory-like (TIML) NK cells exhibit a heightened, tumor-restricted cytotoxicity which is dependent on a higher/faster perforin but not IFN-γ release. Comparative transcriptome analysis reveals that gene expression patterns differ between TIML- and Cytokine-induced memory-like (CIML)-NK cells.