Presence Of IL-22 Research Articles

Inulin diet uncovers complex diet-microbiota-immune cell interactions remodeling the gut epithelium

BackgroundThe continuous proliferation of intestinal stem cells followed by their tightly regulated differentiation to epithelial cells is essential for the maintenance of the gut epithelial barrier and its functions. How these processes are tuned by diet and gut microbiome is an important, but poorly understood question. Dietary soluble fibers, such as inulin, are known for their ability to impact the gut bacterial community and gut epithelium, and their consumption has been usually associated with health improvement in mice and humans. In this study, we tested the hypothesis that inulin consumption modifies the composition of colonic bacteria and this impacts intestinal stem cells functions, thus affecting the epithelial structure.MethodsMice were fed with a diet containing 5% of the insoluble fiber cellulose or the same diet enriched with an additional 10% of inulin. Using a combination of histochemistry, host cell transcriptomics, 16S microbiome analysis, germ-free, gnotobiotic, and genetically modified mouse models, we analyzed the impact of inulin intake on the colonic epithelium, intestinal bacteria, and the local immune compartment.ResultsWe show that the consumption of inulin diet alters the colon epithelium by increasing the proliferation of intestinal stem cells, leading to deeper crypts and longer colons. This effect was dependent on the inulin-altered gut microbiota, as no modulations were observed in animals deprived of microbiota, nor in mice fed cellulose-enriched diets. We also describe the pivotal role of γδ T lymphocytes and IL-22 in this microenvironment, as the inulin diet failed to induce epithelium remodeling in mice lacking this T cell population or cytokine, highlighting their importance in the diet-microbiota-epithelium-immune system crosstalk.ConclusionThis study indicates that the intake of inulin affects the activity of intestinal stem cells and drives a homeostatic remodeling of the colon epithelium, an effect that requires the gut microbiota, γδ T cells, and the presence of IL-22. Our study indicates complex cross kingdom and cross cell type interactions involved in the adaptation of the colon epithelium to the luminal environment in steady state.-dZ131ovLvfeoRbZ-Pz4qeVideo

Evaluation of Triple Fragment Vaccine HSPX (Rv2031c) + PPE44 (Rv2770c) + Mouse IgG1 (Fcγ2a) with Auxiliary Adjuncts IL-22 in Comparison with BCG Vaccine.

Background & Objective:Despite the vaccination with the BCG vaccine, tuberculosis (TB) remains one of the major health problems in the world. The aim of this study was to evaluate our newly designed vaccine using IL-22 as an adjuvant in comparison with the common BCG vaccine. Methods:The gene constructs were cloned into the expression vector of pET28a and then into the recombinant vector of PET28a – HSPX, and PPE44 was transformed into Escherichia coli BL21 (DE3). Finally, the immunogenicity of recombinant proteins with and without BCG and IL-22 in BALB/c mice was investigated.Results:The key cytokines INF-γ and TNF-α were elevated more greatly in BCG immunized group than in PHF immunized group. Immunization with PHF showed a significant increase in IL-4 levels versus the BCG group. Adding IL-22 to the vaccine formulations indicated a tiny increase in IL-4 levels compared to their related vaccine groups.Specific total IgG1 in the experimental groups showed an increase in comparison with control groups, but in the vaccinated groups, no significant differences were observed, and the presence of IL-22 in the vaccine formulations indicated a slight decrease compared with the related mere vaccine groups. Results of specific total IgG2a in the experimental groups revealed that only in the PHF group formulated with IL-22 a significant increase occurs compared with all other experimental groups. Conclusion:It seems that BCG, as the only licensed vaccine for TB infection, could be more potent than a recombinant vaccine in the induction of cellular and humoral immune responses.

Differential effects of Th17 cytokines during the response of neutrophils to Burkholderia cenocepacia outer membrane protein A.

T helper 17 cells are involved in the immunopathology of cystic fibrosis. They play a key role in recruitment of neutrophils, which is the first line of defence against bacteria. Additionally, Burkholderia cenocepacia outer membrane protein A (OmpA) BCAL2958 is considered a potential protective epitope for vaccine development. The present study aimed to investigate the neutrophil response to OmpA in the presence of Th17 cytokines, IL-17 and IL-22 at different times of activation. Neutrophils were isolated from whole blood of healthy volunteers and activated with OmpA in the presence of IL-17, IL-22 or both cytokines together. Supernatant was collected after 1 h, 2 h, 4 h, 8 h, and 12 h. Neutrophil activation was assessed by measuring MPO, TNF-α, elastase, hydrogen peroxide, catalase and NO. The results revealed that the combination of IL-17 and IL-22 cytokines induced the release of NE, catalase, H2O2 and TNF-α from neutrophils activated with Burkholderia OmpA at late stages of activation. However, IL-22 alone or IL-17 alone decreased the myeloperoxidase (MPO), catalase and NE levels at early stages of neutrophil activation. The presence of IL-17 alone led to a significant increase in TNF-α level after 1 h and 12 h. However, the presence of IL-22 alone led to a significant increase in TNF-α level after only 1 h but a significant decrease after 8 h of activation was observed as compared to OmpA stimulated neutrophils. In conclusion, Th17 cytokines IL-17 and IL-22, have differential effects during the neutrophil response to Burkholderia OmpA.

Abstract B26: Novel pro-host role for insulin receptor substrate-1 in lung adenocarcinoma

Abstract Lung cancer is the leading cause of cancer deaths worldwide. In the United States alone, lung cancer accounts for ~160,000 deaths per year while the five-year survival rate remains stagnant at ~15%. Lung adenocarcinoma (ADCA) accounts for over 50% of non-small lung cancer cases. Within this subset, K-ras is the most common activating mutation accounting for ~35% of cases. Kras mutant tumors have remained a largely un-targetable subtype and a better understanding of the signaling pathway involved in the different ADCA subtypes will be necessary for the development of therapeutics. Insulin Receptor Substrate-1 (IRS-1) is a signaling adaptor protein that interfaces with many of the pathways that are activated in lung cancer. We have undertaken a controlled study of this protein within the context of lung ADCA. Analysis of a well-annotated human lung ADCA TMA (n=136) revealed that positive IRS-1 staining provided a 75-month median survival advantage among all ADCA cases (p=0.01) and an 81-month median advantage within the K-ras subtype (p=0.006). EGFR and non-K-ras/non-EGFR cases did not display differences. To focus in on the K-ras subtype, we generated LSL-K-ras/IRS-1fl/fl mice and studied them over a time course. LSL-K-ras/IRS-1-/- mice displayed highly significant early mortality (p<0.0001) and increased tumor burden when compared to LSL-K-ras/IRS-1+/+ controls (p<0.01). Interestingly, IRS-1 loss in tumor cells generated a robust immune response. The bronchial alveolar lavage fluid of LSL-K-ras/IRS-1-/- mice showed increased immune infiltration, most prominently neutrophilic (5 times higher). Significant increases in many immune cell-recruiting chemokines were observed in the LSL-K-ras/IRS-1-/- mice compared to controls including CCL2, CCL3, CCL4, CXCL1, CXCL2 and CXCL5. Surprisingly, an array of IRS-1 silenced A549 cells versus controls displayed no change in CC/CXC chemokine production. We decided to investigate which factor could be missing in our in vitro system that is necessary for activation of this chemokine response. IL-17 and IL-22 producing cells, which are present at sites of tumor in human lung cancer cases and LSL-K-ras mice are known to induce CC/CXC chemokine responses. The JAK/STAT pathway is also implicated in this response. We found that loss of IRS-1 increases pSTAT3 production in vivo and in vitro in the presence of IL-22. Finally, we treated LSL-K-ras/IRS-1-/- and LSL-K-ras/IRS-1+/+ controls with a JAK inhibitor (AZD1480). A significant reduction in tumor burden and chemokine response was observed in both genotypes, with the most pronounced response in the LSL-K-ras/IRS-1-/- group. K-ras mutant, IRS-1-low patients represent ~40% of K-ras mutant lung ADCA patients. The phenotype of this group of patients is counter-intuitive in that IRS-1 typically functions in a pro-growth manner. In the setting of certain oncogenic drivers, however, IRS-1 plays an opposing role to this in which its presence actually maintains homeostasis. This work provides evidence that JAK inhibition may be a viable therapeutic option for these patients as well as describes a novel role for IRS-1 as a mediator of immune cell recruitment in lung adenocarcinoma. Citation Format: Heather E. Metz, Julia Kargl, Stephanie Busch, Kyoung-Hee Kim, McGarry Houghton. Novel pro-host role for insulin receptor substrate-1 in lung adenocarcinoma. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr B26.

Differential effects of t helper-17 cytokines on the functions of granulocytes isolated from schistosoma mansoni-infected patients and healthy individuals

Background and aim The role of T helper-17 lymphocytes in the regulation of the immune response against Schistosoma soluble egg antigens is still controversial. In this study, the in-vitro effects of T helper-17 cytokines [interleukin (IL)-17 and IL-22] on granulocyte functions isolated from Schistosoma- infected patients or healthy individuals were examined. Patients and methods Twenty-seven Schistosoma mansoni- infected patients and 13 healthy individuals from Kasr Al-Aini Viral Hepatitis Center were enrolled in the present study. Granulocytes were isolated from whole blood of patients and controls by Ficoll–Paque density gradient for removal of the mononuclear layer and then lysis of red blood cells. Granulocytes were stimulated in vitro with soluble egg antigen in the presence of IL-17, IL-22, or both. After 24 h, the supernatants were collected for the measurement of tumor necrosis factor (TNF)-α, hydrogen peroxide (H 2 O 2 ), myeloperoxidase (MPO), and nitric oxide (NO) using enzyme-linked immunosorbent assay as surrogate markers for granulocyte functions. Results The results indicated that the presence of IL-17 significantly decreased ( P 2 O 2 , MPO, and NO production by granulocytes isolated from Schistosoma- infected patients. In contrast, in the presence of IL-22 or both IL-17 and IL-22, there were significant increases in the production of H 2 O 2 and TNF-α by granulocytes isolated from Schistosoma- infected patients. Moreover, in the presence of both IL-17 and IL-22, nonsignificant changes were observed in MPO or NO levels compared with those in the control participants. Conclusion IL-17, in contrast to IL-22, inhibited the functional activity of granulocytes isolated from S. mansoni- infected patients. Therefore, neutralization of IL-17 may work as a therapeutic strategy for these patients.

IL-22 capacitates dermal fibroblast responses to TNF in scleroderma

ObjectivesInterleukin (IL) 22 mRNA in systemic sclerosis (SSc) skin and Th22 cells in SSc peripheral blood are increased, but the role of IL-22 in fibrosis development remains poorly understood.MethodsBiopsies were...

Abstract 2354: Insulin receptor substrate-1 regulates immune cell content in lung adenocarcinoma

Abstract Lung cancer is the leading cause of cancer deaths worldwide. In the United States, lung cancer accounts for ∼160,000 deaths per year while the five-year survival rate remains stagnant at ∼15%. A better understanding of the pathobiology for this disease is imperative for development of novel therapeutics to improve mortality rates. Lung adenocarcinoma (ADCA) accounts for over 50% of non-small lung cancer cases. Within this subset, K-ras is the most common activating mutation accounting for ∼35% of cases. Our group previously showed that K-ras activation generates an inflammatory response in lung tumors, mediated largely through the increased production of CC/CXC chemokines by tumor cells. The corresponding influx of neutrophils increased tumor growth, which was associated with degradation of the intracellular protein insulin receptor substrate-1 (IRS-1). Since IRS-1 is generally thought of as a pro-tumor entity, we decided to undertake a controlled study to further elucidate its pro-host role. We performed a study using a well-annotated human lung ADCA TMA (n = 136) and found that positive IRS-1 staining provided a 75-month median survival advantage among all cases (p = 0.01) and an 81-month median advantage within the K-ras subtype (p = 0.006). EGFR and non-K-ras/non-EGFR cases did not display differences. To further dissect the role of IRS-1 in tumor growth we generated LSL-K-ras/IRS-1fl/fl mice and studied them over a time course. LSL-K-ras/IRS-1-/- mice displayed highly significant early mortality (p<0.0001) and increased tumor burden when compared to LSL-K-ras/IRS-1+/+ controls (p<0.01). Surprisingly, IRS-1 loss in tumor cells generated a robust immune response. The bronchial alveolar lavage fluid of LSL-K-ras/IRS-1-/- mice had increased infiltration of macrophages (∼2.5x), neutrophils (∼40x) and lymphocytes (∼5x). Significant increases in many immune cell-recruiting chemokines were observed in the LSL-K-ras/IRS-1-/- mice compared to controls. Many cancers are able to manipulate the host immune response through signaling pathways that interface with IRS-1, including: phosphoinositol 3-kinase (PI3K), extracellular signal regulated kinase (MEK/ERK) and Janus kinase/signal transducer and activator of transcription (JAK/STAT). An array of IRS-1 silenced A549 cells versus controls displayed no change in CC/CXC chemokine production. Evaluation of the tumor microenvironment, however, provided evidence for a role for IL-17 and IL-22 producing cells, which are present at tumor sites in human lung cancer cases and LSL-K-ras mice. We found that loss of IRS-1 increases pSTAT3 production in vivo and in vitro in the presence of IL-22, triggering chemokine release and the generation of a pro-tumor immune response. This is the first description of a pro-host role for IRS-1 in human cancer. This study provides an important marker in a subset (K-ras) of lung ADCA with a very poor prognosis that may be susceptible to JAK/STAT antagonism. Citation Format: Heather Metz, Stephanie E. Busch, Julia Kargl, Mark L. Hanke, Kyoung-Hee Kim, A. McGarry Houghton. Insulin receptor substrate-1 regulates immune cell content in lung adenocarcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2354. doi:10.1158/1538-7445.AM2015-2354

IL-22 is related to development of human colon cancer by activation of STAT3

BackgroundIt has been previously reported that IL-22, one of the cytokines secreted by Th17 cells, demonstrates both a protective and inflammatory promotion effect in inflammatory bowel disease (IBD) through STAT3 signaling activation. We sought to investigate the role of IL-22 expression in colon cancer (CC).MethodsThe expression of IL-22 and related molecules were detected in human CC, the detail function and mechanism of IL-22 were investigated by in vivo and in vitro model.ResultsOur results demonstrated significant upregulation of IL-22 in human CC tumor infiltrated leukocytes (TILs) compared to peripheral lymphocytes. Moreover, our findings demonstrated that IL-22 expression was significantly higher in ulcerative colitis (UC) tissues versus normal colon tissues. Both IL-22 receptor α1 (IL-22RA1) and IL-23 were highly expressed in CC and UC tissues compared to normal controls. TILs exhibiting various IL-22 expression levels isolated from CC patients were demonstrated to enhance tumor growth and metastasis co-transplanted with Hct-116 cells underwent subcutaneous transplantation in mice model. Tumor growth and metastasis was promoted by STAT3 phosphorylation and upregulation of its downstream genes such as Bcl-xl, CyclinD1, and VEGF. In vitro studies confirmed the anti-apoptotic and pro-proliferation effect of IL-22 according to the BrdU cooperation assay and peroxide induced apoptosis analysis with or without the presence of IL-22.ConclusionIn this study we demonstrated that excessive IL-22 in the CC and UC microenvironment leads to tumor growth, inhibition of apoptosis, and promotion of metastasis depend on STAT3 activation.

The involvement of interleukin-22 in the expression of pancreatic beta cell regenerative Reg genes.

BackgroundIn Type 1 diabetes, the insulin-producing β-cells within the pancreatic islets of Langerhans are destroyed. We showed previously that immunotherapy with Bacillus Calmette-Guerin (BCG) or complete Freund’s adjuvant (CFA) of non-obese diabetic (NOD) mice can prevent disease process and pancreatic β-cell loss. This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas.ResultsWe hypothesized that IL-22 was responsible for the increased Reg gene expression in the pancreas. We therefore quantified the Reg1, Reg2, and Reg3δ (INGAP) mRNA expression in isolated pre-diabetic NOD islets treated with IL-22. We measured IL-22, and IL-22 receptor(R)-α mRNA expression in the pancreas and spleen of pre-diabetic and diabetic NOD mice. Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22.ConclusionsWe conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.

IL-22 is involved in atherosclerosis lesions by regulated impaired proliferation ability of oxidized low density lipoprotein treated CRL-1730

Objective To investigate the association between IL-22 and the pathogenesis of coronary artery atherosclerosis(AS).Methods The relative expression of IL-22 mRNA in PBMC from 30 AS patients and 8 patients without any signs of coronary artery stenosis was detected by RT-PCR.Serum IL-22 levels of 22 patients without any signs of coronary artery stenosis and 79 AS patients were detected by ELISA.CRL-1730 cells(human umbilical vein endothelial cells) were stimulated with oxidized low density lipoprotein (ox-LDL) at different dosage for 24 h,and the expression of IL-22R1 was detected by flowcytometry.The proliferation ability of CRL-1730 cells treated with IL-22(20 ng/ml) and/or ox-LDL(100 μg/ml)was measured by MTS assay,and the expression of basic fibroblast growth factor(bFGF) was detected by RTPCR and ELISA.Results Decreased IL-22 expression in PBMC and serum was observed as worsen of AS.The expression of IL-22R1 in ox-LDL treated CRL-1730 cells was increased in dose dependent manner.OxLDL decreased proliferation ability,as well as bFGF expression and releasing,of CRL-1730 cells.This effect of ox-LDL was partially rescued by IL-22.Conclusion IL-22 may have anti-atherosclerosis effect.This effect may be mediated by regulating bFGF expression and endothelial cells proliferation ability in the presence of IL-22. Key words: Atherosclerosis ; Interleukin-22 ; Human umbilical vein endothelial cell ; Oxidized low density lipoprotein ; Proliferation ; Basic fibroblast growth factor