Gene Expression In Immune Cells Research Articles

Mycobacterium tuberculosis-Induced Bronchoalveolar Lavage Gene Expression Signature in Latent Tuberculosis Infection Is Dominated by Pleiotropic Effects of CD4+ T Cell-Dependent IFN-γ Production despite the Presence of Polyfunctional T Cells within the Airways.

Tuberculosis (TB) remains a worldwide public health threat. Development of a more effective vaccination strategy to prevent pulmonary TB, the most common and contagious form of the disease, is a research priority for international TB control. A key to reaching this goal is improved understanding of the mechanisms of local immunity to Mycobacterium tuberculosis, the causative organism of TB. In this study, we evaluated global M. tuberculosis-induced gene expression in airway immune cells obtained by bronchoalveolar lavage (BAL) of individuals with latent TB infection (LTBI) and M. tuberculosis-naive controls. In prior studies, we demonstrated that BAL cells from LTBI individuals display substantial enrichment for M. tuberculosis-responsive CD4+ T cells compared with matched peripheral blood samples. We therefore specifically assessed the impact of the depletion of CD4+ and CD8+ T cells on M. tuberculosis-induced BAL cell gene expression in LTBI. Our studies identified 12 canonical pathways and a 47-gene signature that was both sensitive and specific for the contribution of CD4+ T cells to local recall responses to M. tuberculosis In contrast, depletion of CD8+ cells did not identify any genes that fit our strict criteria for inclusion in this signature. Although BAL CD4+ T cells in LTBI displayed polyfunctionality, the observed gene signature predominantly reflected the impact of IFN-γ production on a wide range of host immune responses. These findings provide a standard for comparison of the efficacy of standard bacillus Calmette-Guérin vaccination as well as novel TB vaccines now in development at impacting the initial response to re-exposure to M. tuberculosis in the human lung.

Antimicrobial peptide, cLF36, affects performance and intestinal morphology, microflora, junctional proteins, and immune cells in broilers challenged with E. coli

This study investigated the effects of an antimicrobial peptide (AMP), cLF36, on growth performance and the histophysiological changes of the intestine in E. coli-challenged broiler chickens. A total number of 360 day old male chicks were randomly assigned to 4 groups of 6 replicates as follows: T1) negative control diet based on corn-soybean meal without E. coli challenge and additives; T2) positive control diet based on corn-soybean meal and challenged with E. coli without any additives; T3) positive control diet challenged with E. coli and supplemented with 20 mg AMP (cLF36)/kg diet; T4) positive control diet challenged with E. coli and supplemented with 45 mg antibiotic (bacitracin methylene disalicylate)/kg diet. Results showed that T3 improved growth performance and the jejunal morphology of E. coli-challenged chickens similar to those of T4. While antibiotic non-selectively decreased the population of ileal bacteria, AMP increased the population of Lactobacillus spp. and decreased harmful bacteria in the ileum of E. coli-challenged chickens. Supplementing E. coli-challenged chickens with AMP improved the gene expression of immune cells and upregulated the expression of tight junction proteins compared to other challenged groups. In conclusion, although cLF36 beneficially affected growth performance and the intestinal morphology of E. coli-challenged chickens similar to those of the antibiotic group, this AMP drastically improved the intestinal microbiome, immune cells, and junctional proteins compared to other E. coli-challenged birds, and can be nominated as an alternative for growth promoter antibiotics.

Toxoplasmosis and abortion: pro- and anti-inflammatory cytokines gene expression of the host immune cells

BackgroundToxoplasmosis during pregnancy is known for inducing variable serious outcomes. In many previous studies, pregnancy was evaluated as a single event while in reality; it has different distinct immunological stages depending on gestational time and possible external factors as infectious agents. A state of immunological balance as well as a state of maternal-fetal tolerance should be established; otherwise, atypical unwelcomed outcomes might occur as a result of such infectionAimThis work aimed to study the gene expression profile of some pro- and anti-inflammatory cytokines in Toxoplasma seropositive casesResultsAll studied cytokines were significantly upregulated in group I, with a remarkable bias towards IFN-γ, denoting a predominant inflammatory state in group I. In group II; tested cytokines were approximately in a balanced state with a little shift towards anti-inflammatory cytokines (IL-10 and TGF-β). A significant positive correlation was detected between pro-inflammatory together (p = 0.002, r = 0.96) and between anti-inflammatory cytokines together (p = 0.0008, r = 0.99) in group II, indicating the harmonized and synchronized expression of both pro- and anti-inflammatory cytokines. In group III, IL-10 was relatively higher with negative correlation with IFN-γ (p = 0.006, r = 0.93), indicating the possible role of IL10 in saving some of the pregnancies’ outcomes in group III.ConclusionThe present study suggests a potential role of these cytokines in the upcoming prognostic or/and therapeutic concerns of toxoplasmosis among pregnant women.

Serum Vitamin D Metabolites and CXCL10 Concentrations Associate With Survival in Dogs With Immune Mediated Disease.

Background: Low vitamin D increases the risk of immune-mediated disease (IMD) in human beings and rodent models. Vitamin D metabolites, particularly 1,25(OH)2D3, modulate gene expression of immune cells and may attenuate immune pathways that drive IMD.Hypothesis/objectives: Our primary hypothesis was that serum 25(OH)D3 and 1,25(OH)2D3, are reduced in patients with IMD and associate with poorer outcomes. We secondarily hypothesized serum 24,25(OH)2D3 would not be associated with disease or outcome. We also measured serum CXCL10 concentrations to determine if an increase occurs in dogs with IMD and in association with poorer survival.Animals: We enrolled dogs diagnosed with IMD (n = 29) and healthy control dogs (n = 9) in the study with informed client consent.Methods: Serum was collected and stored at −80°C until analyses. Serum vitamin D metabolites were measured by LC-MS/MS by an accredited laboratory. A commercially available canine-specific ELISA kit measured serum CXCL10.Results: Serum 25(OH)D3 and 1,25(OH)2D3 were significantly reduced in dogs (n = 25) with IMD. Serum CXCL10 concentrations undetectable in all controls, and were 30 times higher overall in IMD dogs (n = 25; P = 0.004). CXL10 was, however, undetectable in 40% of dogs with IMD. Of the 60% of IMD dogs with increased CXCL10 concentrations, 5/25 had concentrations at the upper limit of quantification. The survival of those five dogs was significantly (P = 0.049) shorter (72 days) than all other dogs with IMD with measured CXCL10 concentrations. The median survival time (MST) for dogs with 25(OH)D3 concentrations ≤ the median was 106 days, while dogs with concentrations of 25(OH)D3 > the median did not achieve an MST.Conclusions and clinical importance: Serum 25(OH)D3 and 1,25(OH)2D3, but not 24,25(OH)2D3 levels are reduced dogs with IMD. Vitamin D metabolites and CXCL10 may be useful prognostic markers and may be targets for adjunct therapy in canine IMD. These data support the future investigation of vitamin D analogs in the treatment of canine IMD.

190 Young Scholar Presentation: Early life fecal microbiome transplantation can enhance growth and performance in neonatal dairy calves

Abstract The gastrointestinal (GI) tract of a neonatal calf undergoes morphological and functional adaptations after birth. The GI tract during this period is influenced by many factors, including bioactive compounds in colostrum. Early colonization of the neonate gut microbiome can fundamentally influence the neonate predisposition to develop immune and metabolic disorders. Fecal microbiome transplantation (FMT) has demonstrated positive effects on the treatment of GI diseases in humans; therefore, we hypothesized that neonatal gut inoculation with a healthy adult gut microbiota via milk replacer can potentially improve gut development and maturation early in life. We aimed to evaluate the effects of performing an early life FMT in neonatal dairy calves using stool samples from on-farm selected adult healthy donors on growth and health performance, blood immunometabolites, and gene expression in immune cells [polymorphonuclear leukocytes cells (PMNL)]. The on-farm selection of the adult donor was based on health and production records at the Dairy Research and Training Facility (DRTF) at South Dakota State University, as well as fecal samples testing negative for Mycobacterium paratuberculosis, Salmonella, and Cryptosporidium. Sixteen healthy newborn Holstein calves (n = 8/trt) housed in individual hutches were used in a randomized complete block design from birth to 7 wk of age. Calves were fed 2.8 L/d of antibiotic-free milk replacer 2×/d during wk 1 to 5, 1×/d on wk 6, and weaned at d 42. Antibiotic-free starter pellets and water were fed ad libitum. Calves were assigned to either a baseline nutritional program (CON) or 1×/d inoculations with 25 g of fecal donor material (FMT) mixed in the milk replacer from 8 to 12 d of age. Individual intakes of milk and pellets were measured daily. Fecal and respiratory scores were recorded daily. Body weight (BW) and withers height (WH) were recorded weekly. Blood samples were collected weekly for immunometabolic profiling and PMNL isolation for gene expression analysis. Data were analyzed using the MIXED procedure of SAS. There was a trend (P = 0.09) for greater BW (50.8 vs 52.7 kg±0.7) in FMT calves. Similarly, there was a trend for greater WH (P = 0.13) in FMT (82.6 vs 83.8 kg±0.49) calves. Starter intake and fecal scores were not affected (P > 0.23) by FMT inoculation. Improvements observed in growth parameters by FMT inoculation are suggestive that neonatal dairy calves may benefit from this approach to enhance gut health and immunity, which might be further explained by blood immunometabolites and PMNL gene expression analysis.

Prospective pilot trial with combination of propranolol with chemotherapy in patients with epithelial ovarian cancer and evaluation on circulating immune cell gene expression

ObjectiveTo determine the feasibility of pharmacologic beta-adrenergic blockade in women with newly diagnosed stage II-IV epithelial ovarian cancer (EOC) throughout primary treatment. MethodsPatients initiated propranolol prior to beginning chemotherapy or surgery. Feasibility was assessed as proportion able to complete 6 chemotherapy cycles while on adrenergic suppression. Descriptive statistics summarized surveys, and paired changes were analyzed using signed rank tests. Random-intercept Tobit models examined immune response. ResultsMedian age was 59.9; 88.5% were stage IIIC/IV; and 38.5% underwent primary debulking. Thirty-two patients were enrolled; 3 excluded because they never took propranolol; an additional 3 didn't meet inclusion criteria, leaving 26 evaluable. Eighteen of 26 (69%), 90% credible interval (CI) of 53–81%, completed 6 chemotherapy cycles plus propranolol (an 82% posterior probability that the true proportion of success is ≥60%). Among the 23 patients with baseline and six month follow up data, overall QOL, anxiety, and depression improved (P < 0.05) and leukocyte expression of pro-inflammatory genes declined (P = 0.03) after completion of therapy. Decrease from baseline of serum IL-6 and IL-8 preceded response to chemotherapy (P < 0.0014). Change from baseline IL-10 preceded complete response. ConclusionUse of propranolol during primary treatment of EOC is feasible and treatment resulted in decrease in markers of adrenergic stress response. In combination with chemotherapy, propranolol potentially results in improved QOL over baseline.

Abstract 2815: Sulindac reverses an immunosuppressive tumor microenvironment associated with obesity-driven metastatic mammary tumors in mice

Abstract Obesity is an established risk factor for several breast cancer (BC) subtypes. Obese BC patients also show poorer response to therapy, increased metastases, and increased mortality. While high levels of inflammation are thought to be a driver for BC metastasis, it is unknown how chronic exposure to low levels of obesity-associated inflammation contributes to BC progression and metastasis. We previously showed that treatment with the non-steroidal anti-inflammatory drug (NSAID) Sulindac (140 ppm mixed into control [10% kcal/fat] or diet induced obesity [DIO, 60% kcal/fat] diets) offset both the pro-growth and pro-metastatic effects of obesity on BC using three different mouse models of metastatic BC - basal-like E0771, and claudin-low metM-Wnt-lung and metM-Wnt-liver. To characterize the effect of Sulindac treatment on the tumor microenvironment, we conducted microarray analysis on E0771 tumors from mice in all four diet groups (Control, Control+Sulindac, DIO, and DIO+Sulindac). Ingenuity Pathway Analysis revealed significant Sulindac-induced differences (q&amp;lt;0.01) in several pathways relating to immune function. Specifically, Th1/Th2 pathways, CD28 signaling, and markers of dendritic cell maturation were significantly downregulated in DIO tumors compared to all other diet groups. Similarly, immune burden calculation using CIBERSORT showed lower levels of CD4+ T memory cells (p&amp;lt;0.05) and higher levels of resting dendritic cells (p&amp;lt;0.001) in DIO tumors compared to all other diet groups. To further explore Sulindac-associated effects on immune and cancer cell gene expression, we performed single-cell RNA-seq on E0771 tumors. Preliminary analyses showed marked differences in gene expression between DIO and DIO+Sulindac tumor cells. Of the top 25 significant (q&amp;lt;0.01) genes upregulated in DIO vs DIO+Sulindac tumor cells, 15 have been previously shown to be drivers of breast cancer progression or metastasis, with only 2 showing tumor suppressive functions. Similarly, 13 of the top 25 genes upregulated in DIO+Sulindac compared to DIO tumor cells have previously shown to have tumor suppressive functions, while only 2 are pro-tumorigenic. Immune analyses showed fewer immune cells in DIO tumors as well as increased pro-tumorigenic or immunosuppressive gene expression in DIO immune cells, while DIO+Sulindac immune cells showed increased markers of immune activation. Finally, T cell receptor (TCR) analysis showed greater TCR diversity in DIO+Sulindac T cells compared to DIO. Taken together, these preclinical findings indicate that obesity-driven mammary tumor progression and metastatic burden are associated with an immunosuppressive tumor microenvironment that is largely reversed by Sulindac treatment. This suggests treatment with NSAIDs like Sulindac should be further assessed as a possible strategy for improving outcomes in obese breast cancer patients. Citation Format: Shannon B. McDonell, Alyssa J. Cozzo, Lydia K. Eisenbeis, Andrew J. Dannenberg, Stephen D. Hursting. Sulindac reverses an immunosuppressive tumor microenvironment associated with obesity-driven metastatic mammary tumors in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2815.

The immune cell landscape in kidneys of patients with lupus nephritis.

Lupus nephritis is a potentially fatal autoimmune disease for which the current treatment is ineffective and often toxic. To develop mechanistic hypotheses of disease, we analyzed kidney samples from patients with lupus nephritis and from healthy control subjects using single-cell RNA sequencing. Our analysis revealed 21 subsets of leukocytes active in disease, including multiple populations of myeloid cells, T cells, natural killer cells and B cells that demonstrated both pro-inflammatory responses and inflammation-resolving responses. We found evidence of local activation of B cells correlated with an age-associated B-cell signature and evidence of progressive stages of monocyte differentiation within the kidney. A clear interferon response was observed in most cells. Two chemokine receptors, CXCR4 and CX3CR1, were broadly expressed, implying a potentially central role in cell trafficking. Gene expression of immune cells in urine and kidney was highly correlated, which would suggest that urine might serve as a surrogate for kidney biopsies.

Transcriptional Regulation in the Immune System: One Cell at a Time.

Transcriptional regulation of cells in the immune system must be strictly controlled at multiple levels to ensure that a proper immune response is elicited only when required. Analysis in bulk, or ensemble of cells, provides a wealth of important information leading to a better understanding of the various molecular steps and mechanisms involved in regulating gene expression in immune cells. However, given the substantial heterogeneity of these cells, it is imperative now to decipher these mechanisms at a single cell level. Here I bring together several recent examples to review our understanding of transcriptional regulation of the immune system via single cell analysis and to further illustrate the immense power of such analyses to interrogate immune cell heterogeneity.

Phase 1 Open-Label, Multicenter Study of First-in-Class RORγ Agonist LYC-55716 (Cintirorgon): Safety, Tolerability, and Preliminary Evidence of Antitumor Activity

Transcription factor retinoic acid receptor-related orphan receptor γ (RORγ) regulates type 17 effector T-cell differentiation and function and is key to immune cell regulation. Synthetic RORγ agonists modulate immune cell gene expression to increase effector T-cell activity and decrease immune suppression. A phase 1 study evaluated the safety and tolerability of LYC-55716 (cintirorgon), a first-in-class, oral, small-molecule RORγ agonist in adults with relapsed/refractory metastatic cancer. Patients received 28-day treatment cycles of oral LYC-55716; dose and dosing regimen were determined according to pharmacokinetic profile and safety. Primary endpoints were safety and tolerability. Secondary endpoints included pharmacokinetics and objective tumor response rate. No dose-limiting toxicities occurred among the 32 enrolled patients who received LYC-55716 150 mg BID to 450 mg BID. Treatment-related adverse events (AE) were primarily grade 1-2 and included diarrhea (n = 11), fatigue (n = 7), anemia (n = 4), decreased appetite (n = 4), and nausea (n = 4). Grade 3 AEs were anemia (n = 2), elevated gamma-glutamyl transferase (n = 1), and hypophosphatemia (n = 1). Pharmacokinetic concentrations achieved levels expected for target gene regulation. Pharmacodynamic results indicated RORγ pathway engagement. Two patients (NSCLC and sarcomatoid breast cancer) had confirmed partial responses; 11 had disease stabilization for 2 to 12 months (6 received >4 months of treatment). These data support the safety and tolerability of LYC-55716 and selection of 450 mg BID dose for a phase 2a study assessing LYC-55716 clinical activity, safety, and biomarkers in patients with NSCLC, head and neck, gastroesophageal, renal cell, urothelial, and ovarian cancers.

Chinese‐German Cooperation Group Tumor Immunology: Another inspiring Meeting in Deidesheim

Chinese‐German Cooperation Group Tumor Immunology: Another inspiring Meeting in Deidesheim

Impact of genetic polymorphisms on human immune cell gene expression

Abstract While many genetic variants have been associated with risk for human diseases, how these variants affect gene expression in various cell types remains largely unknown. To address this gap, the DICE (database of immune cell expression, expression quantitative trait loci (eQTLs), and epigenomics) project was established. Considering all human immune cell types and conditions studied, we identified cis-eQTLs for a total of 12,254 unique genes, which represent 61% of all protein-coding genes expressed in these cell types. Strikingly, a large fraction (41%) of these genes showed a strong cis-association with genotype only in a single cell type. We also found that biological sex is associated with major differences in immune cell gene expression in a highly cell-specific manner. These datasets will help reveal the effects of disease risk-associated genetic polymorphisms on specific immune cell types, providing mechanistic insights into how they might influence pathogenesis (https://dice-database.org).

Vagus nerve stimulation dampens intestinal inflammation in a murine model of experimental food allergy.

BackgroundThe vagus nerve has emerged as an important modulator of the intestinal immune system. Its anti‐inflammatory properties have been previously shown in innate and Th1/Th17 predominant inflammatory models. To what extent the vagus nerve is of importance in Th2 inflammatory responses like food allergy is still unclear. In this study, we therefore aimed to investigate the effect of vagotomy (VGX) and vagus nerve stimulation (VNS), on the development and severity of experimental food allergy.MethodsBalb/C mice were first sensitized with ovalbumin (OVA) in the presence of alum. Prior to oral challenges with OVA, mice were subjected to VGX or VNS. Disease severity was determined by assessing severity and onset of diarrhoea, OVA‐specific antibody production, mast cell number and activity, inflammatory gene expression in duodenal tissue and lamina propria immune cells by flow cytometry analysis.ResultsWhen compared to control mice, VGX did not significantly affect the development and severity of the disease in our model of food allergy. VNS, on the other hand, resulted in a significant amelioration of the different inflammatory parameters assessed. This effect was independent of α7nAChR and is possibly mediated through the dampening of mast cells and increased phagocytosis of OVA by CX3CR1hi macrophages.ConclusionsThese results underscore the anti‐inflammatory properties of the vagus nerve and the potential of neuro‐immune interactions in the intestine. Further insight into the underlying mechanisms could ultimately lead to novel therapeutic approaches in the treatment of not only food allergy but also other immune‐mediated diseases.

MON-LB018 Depot-Specific Differences in Adipose Tissue Morphology with Laron Syndrome

In 1997, our laboratory generated GH receptor gene-disrupted mice (GHR−/−), a mouse model of GH resistance that resembles the human condition termed Laron Syndrome (LS). These mice have provided countless insights into the numerous actions of GH. In particular, adipose tissue (AT) abundance and function have been extensively studied in these mice. Because GH is a major lipolytic hormone, GHR-/- mice have an increase in body fat percentage throughout life, with a notable and disproportionate increase in subcutaneous depots. Many other characteristics of adipose tissue in these mice have been evaluated, including immune cell infiltration, adipokine expression, fibrosis, senescence and gene expression analysis. Consistently, the subcutaneous depot is most dramatically altered by the absence of GH action in GHR-/- mice. While these mice provide an opportunity to routinely analyze multiple AT depots, their relevance to the clinical condition remains unexplored where invasive sampling of multiple AT depots in humans is not typically feasible. In order to begin addressing the clinical relevance of our mouse data, the purpose of this study was to compare AT morphology and adipocyte size in subcutaneous and visceral AT from a female patient with LS undergoing Roux-en-Y gastric surgery to that of the AT data collected from a control female patient. The patient with LS had a BMI of 43.3; the control patient had a comparable BMI of 39.5. Glucose and insulin levels were comparable between the patients, with glucose values of 104 mg/dl and 107 mg/dl, and insulin values of 10 µU/ml and 15 µU /ml for the patient with LS and the control patient, respectively. Samples of subcutaneous AT were taken from the lower abdominal wall and visceral AT from the greater omentum during the surgery. Tissue was immediately frozen or fixed in buffered formalin, paraffin embedded and processed for histological analyses. As our results in mice would suggest, we observed a more significant enlargement in subcutaneous versus visceral adipocytes from the patient with Laron syndrome. In addition, fibrosis appeared to be reduced to a greater extent in the subcutaneous depot. The difference in adipocyte size and fibrosis between the depots was not apparent in the control sample. While results from a single patient with LS compared to a control has obvious limitations, these valuable LS samples do provide initial evidence for a depot-specific alteration in AT in clinical populations comparable to what has been observed for the GHR-/- mice. In future studies, more extensive gene and protein analyses of the frozen tissue will provide additional insight as to the relevance of our GHR-/- data to clinical populations. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. s presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.

A Systematic Review of the Association Between Bereavement and Biomarkers of Immune Function.

Bereavement is associated with increased risk for morbidity and all-cause mortality across epidemiological, meta-analytic, and case-control studies. The body of research examining the association between bereavement and immune function, beginning in 1977, has yet to be reviewed. The current systematic review clarifies these findings, identifies limitations of the diverse literature, and suggests a model and directions for future research. The PRISMA guidelines for systematic reviews were used to synthesize 41 years of research. Compared with a meta-analysis, a systematic review is appropriate given the heterogeneous nature of the studies. We used the National Heart, Lung, and Blood Institute Study Quality Assessment Tool to assess the quality of select studies. Thirty-three publications met inclusion criteria. Most studies support the association between bereavement and maladaptive changes in immune parameters in adults, with mixed findings on bereavement-related changes in cellular immunity. A handful of recent, good-quality studies show bereaved people demonstrate higher levels of systemic inflammation, maladaptive immune cell gene expression, and lower antibody response to vaccination compared with nonbereaved controls. Individual differences in psychological response to bereavement (e.g., depression, grief) seem to influence the association between bereavement and immune function. Research examining the association between bereavement and biomarkers of immune function is heterogeneous in methods and quality. Despite these limitations, there is evidence supporting maladaptive changes in immune function after bereavement. The research area would benefit from longitudinal research with larger sample sizes, advanced immunological methods, and incorporating measures of psychological responses to bereavement. The field is poised to refine and scale up its investigation of this common and important phenomenon.

Identification and Validation of a Tumour Microenvironment-Based Immune Molecular Subgroups for Gastric Cancer: Immunotherapeutic Implications

Background: Immunotherapy could trigger durable response in advanced gastric cancer, but it only benefits a minority of patients. Aims: To propose a robust molecular classification of gastric cancer microenvironment to identify ideal candidates for tailoring effective immunotherapy. Methods: A training cohort of 375 gastric cancer samples with RNAsequencing data were analysed. We virtually microdissected tumour, stromal, and immune cell gene expression patterns employing a nonnegative matrix factorization algorithm. These expression patterns were annotated using immune- and stromal-related gene signatures. Validation of immunogenomic classification was performed across six microarray datasets of 1,406 samples. Results: We found approximately half of gastric cancer samples to have higher immune cell infiltrates, PD-L1 expression, markers of cytolytic activity, and fewer copy number aberrations (all P < 0.05). We termed this group of tumours the Immune Class, which incorporated two components, namely Immune Activation and Immunosuppressive Subtype, according to immunosuppressive or activated microenvironment. Immune Activation Subtype was associated with improved survival in multivariate survival analysis and shared similar genomic characteristics with responders of anti-PD-1 therapy. Immunosuppressive Subtype featured high immune infiltration, stromal enrichment, and transforming growth factor (TGF)-β signalling pathway activation, which might be suitable for antiPD-L1 and anti-TGF-β combined therapy. Conclusions: We proposed and independently validated three reproducible immune molecular subtypes of gastric cancer, which may provide implications for patient selection of immunotherapy. Funding Statement: This work was supported by grants from the Incubating Program for Clinical Research and Innovation of Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University (Grant No. PYMDT-001), Science and Technology Commission of Shanghai Municipality (Grant No. 16441906903), Three-year action plan for Shin Kang of Shanghai (16CR4005A). Declaration of Interests: The authors declare no potential conflicts of interest. Ethics Approval Statement: Not required.

Investigation of indoleamine 2,3-dioxygenase 1 expression in uveal melanoma

The purpose of this study was to investigate indoleamine 2,3-dioxygenase 1 (IDO1) expression and its implications in uveal melanoma (UM). Bioinformatics analysis was performed on microarray data (GSE22138 and GSE27831) from the Gene Expression Omnibus (GEO) database to evaluate IDO1 expression in mRNA level. Ninety-two cases in the database were divided into the IDO1-high group (46 cases) and IDO1-low group (46 cases). Paraffin embedded tumor sections from 27 patients with UM were studied by immunofluorescence. The mRNA results showed that IDO1 expression was inversely correlated with tumor thickness (9.93 ± 3.33 mm in IDO1-high group vs. 11.56 ± 2.38 mm in IDO1-low group) (p = 0.016) and metastatic rate (30.4% in IDO1-high group vs. 69.6% in IDO1-low group, p < 0.001). The IDO1-high group showed higher immune cell gene expression: CD3D (6.56 ± 1.0 vs. 5.46 ± 0.53, p < 0.0001), CD4 (4.72 ± 0.4 vs. 4.2 ± 0.42, p < 0.0001), and CD68 (6.17 ± 1.23 vs. 5.53 ± 0.77, p = 0.015). Further analysis showed that immune-suppressive T regulatory cell genes (CD3D, CD4, IL2RA and FOXP3) were expressed in 67.4% (31/46) cases in the IDO1-high group and 23.91% (11/46) cases in the IDO1-low group. In addition, IDO1 and interferon gamma (IFNG) mRNA expression were strongly correlated (r = 0.70, p < 0.0001). The correlation analysis of different immune checkpoints showed that IDO1 was positively correlated with CD274(PDL1), but not CTLA4 or PDCD1.The disease-free survival (DFS) in the IDO1-high/IFNG-high group was better than that of the IDO1-low/IFNG-low group. The IDO1 immunostaining result showed that 2 cases in 18 UMs with Bruch's membrane (BM) rupture and 7 out of 9 cases without BM rupture were scored high (Grade 2–3) (p = 0.001). Comparing the immune cells staining results between IDO1-high group and IDO1-low group, higher percentage of patients in the former group had high levels of T cells and macrophages infiltration, but only the difference in macrophage was statistically significant (CD68, 77.78% vs. 27.78%, p = 0.04). The analysis based on GEO data and the result from immunostaining study are consistent with each other. In conclusion, the expression of IDO1 is probably induced by IFNγ from infiltrated immune cells in UM. BM rupture is an important indicator of IDO1 expression level and distribution pattern. The complex role of IDO1 may limit its therapeutic effect in UM.

Social status alters chromatin accessibility and the gene regulatory response to glucocorticoid stimulation in rhesus macaques

Low social status is an important predictor of disease susceptibility and mortality risk in humans and other social mammals. These effects are thought to stem in part from dysregulation of the glucocorticoid (GC)-mediated stress response. However, the molecular mechanisms that connect low social status and GC dysregulation to downstream health outcomes remain elusive. Here, we used an in vitro GC challenge to investigate the consequences of experimentally manipulated social status (i.e., dominance rank) for immune cell gene regulation in female rhesus macaques, using paired control and GC-treated peripheral blood mononuclear cell samples. We show that social status not only influences immune cell gene expression but also chromatin accessibility at hundreds of regions in the genome. Social status effects on gene expression were less pronounced following GC treatment than under control conditions. In contrast, social status effects on chromatin accessibility were stable across conditions, resulting in an attenuated relationship between social status, chromatin accessibility, and gene expression after GC exposure. Regions that were more accessible in high-status animals and regions that become more accessible following GC treatment were enriched for a highly concordant set of transcription factor binding motifs, including motifs for the GC receptor cofactor AP-1. Together, our findings support the hypothesis that social status alters the dynamics of GC-mediated gene regulation and identify chromatin accessibility as a mechanism involved in social stress-driven GC resistance. More broadly, they emphasize the context-dependent nature of social status effects on gene regulation and implicate epigenetic remodeling of chromatin accessibility as a contributing factor.

Effects of Millimolar Steady-State Hydrogen Peroxide Exposure on Inflammatory and Redox Gene Expression in Immune Cells from Humans with Metabolic Syndrome.

Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) can exert opposed effects depending on the dosage: low levels can be involved in signalling and adaptive processes, while higher levels can exert deleterious effects in cells and tissues. Our aim was to emulate a chronic ex vivo oxidative stress situation through a 2 h exposure of immune cells to sustained H2O2 produced by glucose oxidase (GOX), at high or low production rate, in order to determine dissimilar responses of peripheral blood mononuclear cells (PBMCs) and neutrophils on ROS and cytokine production, and mitochondrial dynamics-related proteins, pro/anti-inflammatory and anti-oxidant gene expression. Immune cells were obtained from subjects with metabolic syndrome. H2O2 at low concentrations can trigger a transient anti-inflammatory adiponectin secretion and reduced gene expression of toll-like receptors (TLRs) in PBMCs but may act as a stimulator of proinflammatory genes (IL6, IL8) and mitochondrial dynamics-related proteins (Mtf2, NRF2, Tfam). H2O2 at a high concentration enhances the expression of pro-inflammatory genes (TLR2 and IL1β) and diminishes the expression of mitochondrial dynamics-related proteins (Mtf1, Tfam) and antioxidant enzymes (Cu/Zn SOD) in PBMCs. The GOX treatments produce dissimilar changes in immune cells: Neutrophils were more resistant to H2O2 effects and exhibited a more constant response in terms of gene expression than PBMCs. We observe emerging roles of H2O2 in mitochondrial dynamics and redox and inflammation processes in immune cells.

Development of a Nano DDS for Cancer Immunotherapy Based on Llipid Nanoparticles

The appearance of immune checkpoint inhibitors has been a major turning point in cancer therapy. The success of immune checkpoint therapy has revolutionized the field of cancer therapy, and immunotherapy has joined the cancer treatment ranks as a pillar. To induce effective anti-tumor immune responses, it is necessary both to enhance the activity of immune cells and to block immune suppression by tumor cells. Carrier type drug delivery systems based on nanobiotechnology (nano DDS) represent a potentially useful technology for efficiently achieving both: enhancement of the activity of immune cells and blocking immune suppression. It has become clear that nano DDS can improve the practical utility of a wide variety of immune functional molecules and thus regulate drug kinetics and intracellular dynamics to improve drug efficacy and reduce side effects. We have been in the process of developing a nano DDS for the enhancement of cancer immunotherapy. A nano DDS encapsulating an agonist of a simulated interferon gene pathway greatly enhanced the activity of the agent's antitumor immune response. To block immune suppression, we successfully developed a small interfering RNA loaded into a nano DDS which regulates gene expression in immune cells. In this review, we summarize our recent efforts regarding cancer immunotherapy using nano DDS.