Interferon-gamma Gene Expression Research Articles

Upregulation of interferon gamma gene expression among asymptomatic malaria adults in Bayelsa State, Nigeria

Despite concerted efforts, malaria remains a global public health problem. Treatment for asymptomatic malaria will reduce the health care burden of malaria. However evidence-based data for the presence of systemic pathology in asymptomatic malaria, which might inform treatment remains limited. The present study investigated the current prevalence of asymptomatic malaria in Bayelsa State, Nigeria and differences in interferon gamma (IFNγ) mRNA expression between asymptomatic malaria individuals (AMIs) and uninfected individuals (UIs), in relation to ABO blood groups and hemoglobin genotype (Hb) variants. A cross-sectional design was employed to randomly select 146 non-febrile participants from Sagbama LGA, Bayelsa State. Plasmodium falciparum infection was determined using high-resolution melting analysis and participants were grouped into UIs and AMIs. Hb, blood groups, and IFNγ mRNA expression were determined using electrophoresis, agglutination, and real-time PCR method respectively. Logistic regression and t-test were used to determine significant differences between groups at p < 0.05. The current prevalence of asymptomatic malaria in Bayelsa State was found to be 89.73 %. Age and sex were associated with asymptomatic malaria (p < 0.05). Plasmodium falciparum DNA melt curves for AMIs aligned at 83.89 ± 0.34 °C, while the amplification cycle thresholds were below 30 cycles. Compared with UIs, the gene expression of IFNγ mRNA was significantly (p < 0.001) higher among AMIs. Also, among AMIs those with A, B, and AB blood groups expressed significantly higher levels of IFNγ mRNA compared with blood groups O AMIs. Furthermore, AMIs with HbAA expressed significantly higher levels of IFNγ mRNA compared with HbAS AMIs. The novelty of the present study is the demonstration of the existence of systemic pathology in asymptomatic malaria and the demonstration that the intensity of systemic pathology is dependent on blood types and haemoglobin genotype variants. Findings of the present study have implications for policy creation towards asymptomatic malaria treatment.

Effect of diode pumping solid state laser with wavelength 589 nm on gene expression of interlukine-2 and interferon-γ in human T-lymphocytes.

To evaluate the impact of laser irradiation of human T-cell lymphocyte in culture to determine the photobiomodulatory effects of such irradiation on gene expression, and the release of growth factors and cytokines. The experimental study was conducted from November 2021 to April 2022 at the Postgraduate Anatomy Laboratory, Mustansiriyah University, Baghdad, Iraq, and comprised human blood samples T-cell lymphocytes were extracted and irradiated, followed by the extraction of ribonucleic acid to assess the effect of laser on gene expression of cytokines, interlukine-2 and interferon-gamma. A 589nm yellow light low-level laser beam from a diode pumping solid state laser was used at doses 30J/cm2, 50J/cm2 and 70J/cm2 for an exposure time of 15 minutes. Reverse transcriptase-polymerase chain reaction was used to measure gene expression. Data was analysed using GraphPad Prism 9. Cellular gene expression of the mediators changed significantly in response to laser irradiation at a variety of dosage parameters, and these effects depended on the wavelength and radiation exposure (p<0.05). The laser light may photobiomodulate the production of cytokines by human cells in vitro.

Abstract 5182: Exploiting tumor RNA-sequencing data for prediction of immune checkpoint inhibition response

Abstract Immune checkpoint inhibitors (ICI) have become the standard of care as a first- or second-line systemic treatment for patients with various cancer types. Unfortunately, many patients do not respond to this treatment and the occurrence of immune-related adverse events varies widely. There is an urgent need for robust predictive biomarkers for ICI response to identify patients with likely clinical benefit from this costly treatment. Several biomarkers, aimed at predicting response to ICI, have been proposed, including PD-L1 expression, tumor mutation burden (TMB), infiltration of cytotoxic T-cells in the tumor, Microsatellite Instability (MSI) and expression of various immune gene signatures. However, these individual biomarkers have suboptimal performance and multiple complementary omics technologies are required to properly quantify them. Integration of these multi-omic biomarkers has proven valuable in increasing the accuracy and robustness of ICI response prediction.We have developed a computational pipeline that determines the expressed mutation burden (eTMB), MSI status, fraction of infiltrating immune cells and various immune gene expression signatures directly from the RNA-sequencing profile of the tumor. Each biomarker is quantified using a dedicated algorithm that applies machine learning (eTMB and MSI) or computational deconvolution (infiltrating immune cells) and integrates external data sources to maximize performance. Algorithm performance has been validated on large cohorts of tumor RNA-sequencing data with matching gold standard quantification of said biomarkers. As a proof of concept, we applied our pipeline to tumor RNA-sequencing data of 45 gastric cancer patients treated with pembrolizumab. Responders showed significantly higher eTMB scores and were enriched among the MSI-H patients. Fractions of CD8 T-cells and M1 Macrophages, together with interferon gamma and cytotoxic T-cell gene expression signatures, were significantly increased in responders. Notably, integrating these biomarkers into a single prediction model improved prediction of response to checkpoint inhibition therapy compared to predictions based on eTMB or MSI alone. Taken together, our approach enables the quantification of various ICI response biomarkers from a single omics layer (RNA-sequencing) and can contribute to ICI response prediction. Citation Format: Pieter Mestdagh, Pieter-Jan Van Dam, Frederick De Baene, Carolina Fierro, Elise Van Hoof, Emmanuel Rivière, Hanne Dangreau, Reindert Van Cauwenberge, Jo Vandesompele. Exploiting tumor RNA-sequencing data for prediction of immune checkpoint inhibition response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5182.

Demonstrating the immunostimulatory and cytokine-augmentation effects of bacterial ghosts on natural killer cells and Caenorhabditis Elegans.

The potential of bacteria-based immunotherapy lies in its ability to inherently enhance immune responses. However, the "liveness" of bacteria poses risks of bacterial escape, nonspecific immuno-stimulation, and ethical concerns, limiting their acceptability in immunotherapy. In this scenario, nonliving empty bacterial-cell envelopes, named bacterial ghosts (BGs), have emerged as immuno-stimulants with the potential to side-step the limitations of live bacterial therapies. This study demonstrates the capability of BGs in modulating the functionality of NK-92 cells and Caenorhabditis elegans (C. elegans), as well as perform as cytokine-therapy adjuvants. BGs were obtained through a pH-driven culture method, and were validated for their structural and chemical integrity via electron microscopy and spectroscopy. In NK-92 cells, BGs have shown significant immuno-stimulation by boosting the gene-expression of perforin, granzyme-B, Fas-L, and interferon-gamma by factors of 3.5-, 1.5-, 12.5-, and 8.6-folds, respectively. Combined BG and IL-12 treatment yielded a notable 10.2-fold increase in interferon-gamma protein expression in 24 h. The BGs also significantly influenced the innate immune response in C. elegans through the upregulation of lysozyme genes viz., ilys-3 (8.8-fold) and lys-2 (3.1-fold). Our investigation into the impact of BGs on natural killer cells and C. elegans highlights its potential as a valid alternative approach for new-age immunotherapy and cytokine augmentation.

Effect of Acute and Cumulative Stress on Gene Expression in Mammary Tissue and Their Interactions with Physiological Responses and Milk Yield in Saanen Goats

This study addresses the hypothesis that different acute stressors can cumulatively decrease milk yield. In fact, in a time of global warming, the impact of environmental stress and farm management practices on milk production remains unclear. In this context, our objective was to investigate the effect of acute and cumulative stress on gene expression in mammary tissue and their interactions with physiological responses and milk yield in Saanen goats. Thirty lactating goats were subjected to two treatments: (1) control (CT), in which goats were maintained following a habitual routine under comfort conditions; (2) stress (ST), in which the goats were subjected to different types of environmental stress: heat stress, adrenocorticotropic hormone administration, hoof care management, and exposure to rain. These stressors were performed sequentially, with one stress per day on four consecutive lactation days, to evaluate their effect on milk quality and milk yield. Our results showed that compared to CT goats, cumulative stress increased the gene expression of glucocorticoid receptor (GR), interferon-gamma (IFN-γ), superoxide dismutase (SOD), and catalase (CAT) in mammary tissue, which are indicators of cortisol action, inflammatory response, and antioxidant enzymes. Furthermore, the acute challenges imposed on ST goats changed their rectal temperature and respiratory frequency and increased cortisol, glucose, cholesterol, triglycerides, and high-density lipoprotein release in plasma when compared to CT goats. Although these physiological and metabolic responses restore homeostasis, ST goats showed lower milk yield and higher somatic cell count in milk than CT goats. In conclusion, the results confirmed our initial hypothesis that different acute stressors cumulatively decrease the milk yield in Saanen goats.

Relation between interferon-gamma gene expression and its serum level with thyroid hormone status and Ferritin level in Iraqi sickle cell anemia patients

The hereditary hemoglobinopathy known as sickle cell disease is characterized by abnormal hemoglobin synthesis, hemolytic anemia, and intermittent obstruction of tiny blood arteries. The current investigation aims to study the relationship between interferon-gamma gene expression and serum levels with thyroid hormone status and Ferritin levels in sickle cell anemia patients. Blood samples were collected from 50 patients suffering from SCA and 50 healthy volunteers as a control group. The results of the relation of IFN-γ serum level with T3, T4 and TSH levels showed a positive relationship between IFN-γ serum and TSH levels. At the same time, an inverse association to T3 and T4 levels with high significance also showed high IFN-γ gene expression (10.78 ±5.06 Fold) in the patient group as compared with control (2.079 ±0.52 Fold) at a significant difference, as well as the results found acquire strong positive association between IFN-γ serum level and IFN-γ mRNA expression in the patient group compared with the control group. The current study concluded that there was an inverse association among T3, T4 and IFN-γ serum with high significance, as high gene expression of IFN-γ, and a strong positive association between IFN-γ serum level and IFN-γ mRNA expression. Keywords: IFN-γ, SCA, Ferritin, TSH, T3, T4.

The Effects of Mesenchymal Stem Cells on the Gene Expression of TGF-beta and IFN-gamma in Patients with Rheumatoid Arthritis.

The therapeutic and immunomodulatory potential of mesenchymal stem cells (MSCs) in rheumatoid arthritis (RA) has attracted considerable scientific attention in recent decades. This study aimed to evaluate the expression of genes encoding interleukin (IL)4 and IL10, as well as interferon-gamma (IFNG) and transforming growth factor beta (TGFB1) in refractory RA patients following intravenous injection of autologous bone marrow-derived MSCs (BM-MSCs). This study was registered in Iranian Registry of Clinical Trials (IRCT) (2015102824760N1) and ClinicalTrials.gov (identifier: NCT03333681). Blood samples were taken from 13 patients before and 1 and 6 months after the MSC injection to evaluate the clinical manifestations, paraclinical factors, and expression of IL4, IL10, IFNG, and TGFB1 genes employing the SYBR Green real-time reverse-transcriptase polymerase chain reaction (RT-PCR) technique. There was a significant increase in the expression of TGFB1 at 1 and 6 months after the MSC injection compared to that in the baseline, while the expression of IL4 and IL10 did not change significantly. On the other hand, the expression of IFNG increased significantly after 1 month but decreased significantly at 6 months compared to 1 month after the intervention. Nevertheless, it showed no significant decrease compared to the baseline. A significant decrease was observed for the expression of IFNG 6 months after the injection compared to that after 1 month, which was in concordance with the rise in the expression of the TGFB1 gene. A significant change in the gene expression of TGFB1 and IFNG in our study wasconsistent with the amelioration of clinical manifestations, suggesting a mechanism of action for MSCs in the treatment of RA.

Effect of gamma-irradiated honey bee venom on gene expression of inflammatory and anti-inflammatory cytokines in mice.

In this study, the effect of gamma-irradiated honey bee venom (doses of 0, 2, 4, 6, and 8kGy, volume of 0.1ml and concentration of 0.2mg/ml) was evaluated on the reduction of allergen compounds and the gene expression of inflammatory and anti-inflammatory cytokines in mice. Hence, edema activity induced by the bee venom irradiated at 4, 6, and 8kGy was reduced, compared with the control group and that irradiated at 2kGy. In contrast, the paw edema induced by the bee venom irradiated at 8kGy increased, compared with 4 and 6kGy. At all the time periods, there was a significant decrease in the gene expression of interferon gamma (IFN-γ), interleukin 6 (IL-6), and interleukin 10 (IL-10) in the bee venoms irradiated at 4, 6, and 8kGy, compared with the control group and that irradiated at 2kGy. In contrast, there was an increase in the gene expression of IFN-γ and IL-6 in the bee venom irradiated at 8kGy, compared with those irradiated at 4 and 6kGy. Therefore, gamma irradiation at 4 and 6kGy reduced the gene expression of cytokines at each time period by decreasing the allergen compounds of honey bee venom.

Hepatoprotective effect of nanoniosome loaded Myristica fragrans phenolic compounds in mice-induced hepatotoxicity.

In this study, nanoniosome-loaded Myristica fragrans' (MF) phenolic compounds (NLMP) were synthesized and characterized for their physical properties, and hepatoprotective effects on mice with liver toxicity induced by L-asparaginase (LA) injection. According to the results, NLMP has a spherical shape with a 263 nm diameter, a zeta potential of -26.55 mV and a polydispersity index (PDI) of 0.192. The weight and feed intake of mice induced with hepatotoxicity were significantly (p≤ 0.05) increased after they were treated with NLMP (2.5mg/kg body weight of mice). In addition, the blood levels of triglyceride (TG), cholesterol (Chol), liver enzymes (aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP)) and total bilirubin were significantly (p≤ 0.05) decreased. A significant increase (p≤ 0.05) in the blood levels of the antioxidant defence system (glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT)) were also reported after NLMP treatment. NLMP was also led to a significant decrease (p≤ 0.05) in inflammatory-related gene expression of inducible nitric oxide synthase (iNOS) and Interferon-gamma (IFN-γ) in the liver, as well as a meaningful (p≤ 0.05) increase in the expression of SOD as an antioxidant status biomarker. Consequently, the NLMP is recommended as a potential dietary supplement to alleviate the symptoms of LA-induced hepatotoxicity.

Adjuvant dabrafenib plus trametinib (D + T) versus placebo in patients with resected stage III BRAFV600-mutant melanoma: Updated 5-year distant metastases-free survival (DMFS) analysis of COMBI-AD.

9563 Background: DMFS is an important endpoint for patients with stage III cutaneous melanoma, as delaying or preventing systemic disease is associated with improved clinical and patient-reported outcomes. Prior results from the phase 3 COMBI-AD trial (NCT01682083) showed 5-year DMFS rates of 65% with adjuvant D + T vs 54% with placebo (PBO; hazard ratio [HR] = 0.55; 95% CI: 0.44-0.70). An analysis of DMFS by AJCC-7 stages IIIA-C suggested a similar benefit of D + T vs PBO regardless of stage (Dummer R et al. N Engl J Med. 2020). Here, we report 5-year DMFS rates by AJCC-8 stages IIIA-D, other prognostic subgroups, and results of a regression tree analysis with DMFS. Methods: Patients with resected AJCC-7 stage III BRAFV600E/K-mutant melanoma were randomized to either D (150 mg twice daily) + T (2 mg once daily) or 2 matched PBOs for 12 months. Primary endpoint was relapse-free survival (RFS); DMFS was a secondary endpoint. Kaplan-Meier survival analyses were performed to assess the long-term benefits for DMFS rates with D + T vs PBO. The regression tree analysis (data cutoff: 5 years) for all patients (N = 870) evaluated potential prognostic/predictive factors of long-term DMFS including baseline age, sex, region, BRAF mutation type, body mass index, lactate dehydrogenase levels, ECOG, T and N categories, histology, primary tumor ulceration, treatment type, number of lymph nodes with metastases, tumor mutational burden, and interferon-gamma gene expression signature (IFN-γ GES). Results: At 5 years, DMFS rates were higher for patients with AJCC-8 stages IIIB-D disease receiving adjuvant D + T vs PBO (table). Five-year DMFS rates also favored D + T vs PBO in subgroups of patients with microscopic or macroscopic lymph node involvement (table) and those with or without primary tumor ulceration and/or in-transit metastases. A regression tree revealed T and N stage, treatment type, and IFN-γ GES as important variables defining 5-year DMFS subgroups. Conclusions: In this retrospective analysis, adjuvant D + T provided long-term DMFS benefit vs PBO in stage IIIB-D patients with resected BRAFV600E/K-mutant melanoma. Key clinical and patient factors impacting DMFS were similar to prior RFS findings (ESMO 2021; Robert C et al. Ann Oncol. 2021) and included T and N stage, treatment type, and IFN-γ GES. These results further validate the robust long-term clinical benefit of adjuvant D + T for patients with melanoma. Clinical trial information: NCT01682083. [Table: see text]

Supplementation of broiler chicken diets with bovine lactoferrin improves growth performance, histological parameters of jejunum and immune-related gene expression.

The aim of this study was to investigate the influence of dietary supplementation of bovine lactoferrin (bLF) on growth performance, carcass traits, histomorphology of jejunum, immune function and hepatic and splenic gene expression of interferon-gamma (IFN-γ) and interleukine-2 (IL-2) in broiler chickens. A total of 240 one-day-old Ross 308male broiler chickens were randomly allotted into six dietary treatments with four replicate pens (10 chicks per pen) and fed experimental diet in 3 feeding phases (starter: d 0-10, grower: d 11-24 and finisher: d 25-42). The experimental treatments were (1) corn-soya bean meal-based basal diet (control), (2-5) basal diet supplemented with 200, 400, 600, 800mg/kg bLF, respectively, and (6) basal diet supplemented with 200mg/kg oxytetracycline (OTC). The average body weight gain (ABWG) of broilers fed 800mg/kg bLF was 8.48% higher than those fed a corn-soybean meal-based diet during the starter period (d 0-10) (linear effect, p=0.002; quadratic effect, p=0.24). Average daily feed intake (ADFI) and the feed conversion ratio (FCR) were not affected (p>0.05) by bLF supplementation. At 42days of age, the breast meat percentage and carcass yield of broilers fed 800mg/kg bLF compared with the control group significantly increased by 9.51% and 6.03% respectively (p<0.05). Compared with the chicks fed the control diet, the chicks fed diets supplemented with bLF had higher villus height, muscle thickness and villus surface area (p>0.05). Dietary bLF inclusion increased the total immunoglobulin (IgT) titre against sheep red blood cells (SRBCs) antigen (linear effect, p=0.031; quadratic effect, p=0.035) and improved the phytohaemagglutinin-P (PHA-P)-skin test of broilers. Compared with the control, bLF enhanced the gene expression of IFN-γ in spleen (p=0.048, linear effect, p=0.009; quadratic effect, p=0.093) and liver (p=0.012, linear effect, p=0.008; quadratic effect, p=0.01) and IL-2 expression in spleen (p=0.021, linear effect, p=0.026; quadratic effect, p=0.103). The bLF supplementation had no effect on IL-2gene expression in liver (p>0.05, linear effect, p=0.213; quadratic effect, p=0.159). In conclusion, we found that supplementation of broiler diets with 800mg/kg bLF can improve the growth performance, carcass yield, cell-mediated and antibody-mediated immune responses and enhance the IL-2 and IFN-γ gene expression of broilers.

Use of paclitaxel carried in lipid nanoparticles to treat aortic allograft transplantation in rats.

The aim of this study was to test whether lipid core nanoparticles loaded with paclitaxel (LDE-PTX) protect rat aortic allograft from immunological damage. Fisher and Lewis rats were used differing in minor histocompatibility loci. Sixteen Lewis rats were allocated to four-animal groups: SYNG (syngeneic), Lewis rats receiving aorta grafts from Lewis rats; ALLO (allogeneic), Lewis rats receiving aortas from Fisher rats; ALLO+LDE (allogeneic transplant treated with LDE), Lewis rats receiving aortas from Fisher rats, treated with LDE (weekly injection for 3 weeks); ALLO+LDE-PTX (allogeneic transplant treated with LDE-PTX), Lewis rats receiving aortas from Fisher rats treated with LDE-PTX (4 mg/kg weekly for 3 weeks). Treatments began on transplantation day. Thirty days post-transplantation, SYNG showed intact aortas. ALLO and ALLO+LDE presented intense neointimal formation. In ALLO+LDE-PTX, treatment inhibited neointimal formation; narrowing of aortic lumen was prevented in ALLO and ALLO+LDE. LDE-PTX strongly inhibited proliferation and intimal invasion by smooth muscle cells, diminished 4-fold presence of apoptotic/dead cells in the intima, reduced the invasion of aorta by macrophages and T-cells and gene expression of pro-inflammatory tumour necrosis factor-alpha (TNFα), interferon gamma (IFNγ) and interleukin-1 beta (IL-1β). LDE-PTX was effective in preventing the vasculopathy associated with rejection and may offer a potent therapeutic tool for post-transplantation.

The Effects of High-intensity Interval Training and Probiotic Consumption on Interleukin-10 and Interferon-gamma Gene Expression of Gut Tissue in an Animal Model of Fatty Liver

Aims: Non-Alcoholic Fatty Liver Disease (NAFLD) is the most common chronic liver disease, often reported in patients with inflammatory bowel disease. This study aimed to investigate the effects of High-Intensity Interval Training (HIIT) and probiotic use on the expression of interleukin-10 and interferon-gamma genes in intestinal tissue in an animal model of fatty liver. Methods &amp; Materials: In this experimental study, 40 male Wistar rats (weighing 200-250 g) were selected. The study rats were randomly divided into 5 groups, including healthy control, fatty liver, fatty liver + HIIT, fatty liver + probiotic, and fatty liver + HIIT + Probiotics. Oral tetracycline was administered at a dose of 140 mg/kg of body weight (as a solution in 2 mL of water) for 7 days with fatty liver. HIIT was performed on tape 5 times a week for 5 weeks. The obtained data were analyzed using One-way Analysis of Variance (ANOVA) and Tukey post-hoc test at the significant level of P&lt;0.05. Findings: The present research results suggested that the expression of interleukin-10 and interferon-gamma genes in the intestinal tissue of steatosis mice in the fatty liver group was significantly higher than in the healthy control group (P=0.001). HIIT and probiotic use significantly reduced the expression of interleukin-10 and interferon-gamma genes in the intestinal tissue, compared to the fatty liver group (P=0.001). Conclusion: According to the obtained data, HIIT and probiotic consumption can help modulate the expression of genes involved in the innate immune system of intestinal tissue caused by fatty liver.

Effect of dietary supplementation of potential probiotic Weissella confusa on innate immunity, immune‐related genes expression, intestinal microbiota and growth performance of rainbow trout ( Oncorhynchus mykiss )

Rainbow trout, Oncorhynchus mykiss (average initial weight: 115 ± 2.6 g), were fed with diets containing different levels of Weissella confusa (0 [control], 1.5 × 107 [T1], 3 × 107 [T2] and 4.5 × 107 [T3] CFU/g) for 2 months. At the end of the experiment, innate immunity, expression of immune-related genes, intestinal microbiota and growth performance of the fish were investigated. A significant increase (p < .05) was seen in the T2 group compared to other groups in terms of lysozyme activity. In case of alternative complement activity (ACH50), a significant increase (p < .05) was also observed in the T1 and T2 treatment groups compared to the other groups. The obtained data from the gene expressions showed a significant up-regulation of tumour necrosis factor-alpha (TNF-α) and interleukin 8 (IL-8) in W. confusa-fed fish (p < .05), while interferon gamma (IFN-γ) gene expression was only significantly up-regulated in T2. However, the expression of interleukin 10 (IL-10) was down-regulated in the supplemented treatments compared to the control group. Microbiological evaluation revealed that feeding with the W. confusa-supplemented diet significantly enhanced the lactic acid bacteria (LAB) levels (p < .05). Total bacterial count (TBC) was significantly increased in T2 compared to the control group (p < .05). Also, the growth performance of supplemented treatments remarkably improved compared to the control group (p < .05). These results confirmed the beneficial effects of W. confusa as host-derived potential probiotic in rainbow trout with the appropriate inclusion being 3 × 107 CFU/g.

The Antimicrobial Peptide Cecropin AD Supplement Alleviated Soybean Meal-Induced Intestinal Inflammation, Barrier Damage, and Microbial Dysbiosis in Juvenile Turbot, Scophthalmus maximus

This study aimed to investigate the benefit of dietary cecropin AD (CAD) on the intestinal health of turbot (Scophthalmus maximus) fed diets with a high level of soybean meal. A 12-week feeding trial was conducted with four isonitrogenous and isolipidic diets: a fishmeal-based diet (FM), a diet replacing 40% fish meal protein of FM diet with soybean meal protein (SBM), and the SBM diet supplemented with 0.5 g/kg (C1) and 1.0 g/kg (C2) CAD, respectively. The results of histology of distal intestine (DI) showed that turbots fed the SBM diet exhibited distinct symptoms of enteritis. However, fish fed diets with CAD supplementation kept the normal appearance of the DI which was similar to that in FM group. Compared with the SBM group, diet with CAD supplementation significantly decreased the gene expression of intestinal pro-inflammatory cytokines tumor necrosis factor-α (tnf-α), interleukin-1β (il-1β), interferon-gamma (ifn-γ), and nuclear factor-kappa B p65 (nf-κb p65), while up-regulated the gene expression of intestinal tight junction proteins claudin-3, claudin-4, occludin, and zonula occludens-1 (zo-1). Besides, diet C1 shaped the intestinal microbiota profile toward an anti-inflammatory phenotype represented by the increased abundance of Blutia, Firmicutes/Bacteroides ratio, and decreased Prevotellaceae. In conclusion, dietary CAD could positively modulate the intestinal health of turbot from the impairment induced by soybean meal, which expands its application to help fish better adapt to the increasing plant protein level in aquafeed.

Probiotic Lactobacilli Enhance Immunogenicity of an Inactivated H9N2 Influenza Virus Vaccine in Chickens.

Avian influenza viruses (AIVs) infect a wide range of hosts, including humans and many avian species. Efforts have been made to control this pathogen in chickens using vaccination programs, but that has been met with varying degrees of success. Therefore, identification of more efficacious vaccination strategies is warranted. This study was undertaken to investigate the potential effects of probiotics on the immunogenicity of a beta-propiolactone-whole inactivated virus (WIV) vaccine of H9N2 subtype adjuvanted with the Toll-like receptor-21 ligand, CpG oligodeoxynucleotides 2007 (CpG). Eighty-four 1-day-old White Leghorn layers were allocated into six groups. Two out of six groups received a mixture of probiotic Lactobacillus spp. (PROB) biweekly from days 1 35 of age. Chickens were intramuscularly vaccinated with WIV either alone or adjuvanted with AddaVax™ (WIV+Add) or CpG (WIV+CpG), and one group received saline (phosphate-buffered saline). Primary and secondary vaccinations occurred at days 14 and 28 of age, respectively. The results revealed that the group that received probiotics and was vaccinated with CpG-adjuvanted WIV H9N2 vaccine had higher hemagglutination inhibition titers than the other treatment groups at days 14 and 21 postprimary vaccination. Probiotics did not induce higher IgM or IgY titers in chickens receiving the WIV vaccine only. Concerning their effect on cell-mediated immune responses, probiotics enhanced interferon-gamma (IFN-γ) gene expression and significantly increased secretion of IFN-γ protein by splenocytes in chickens vaccinated with CpG-adjuvanted WIV H9N2. Together, these findings suggest the use of probiotics to enhance the immunogenicity of CpG-adjuvanted WIV H9N2 vaccines. Additional studies are required to better understand the specific interactions between probiotics and the gut microbiota and different types of cells of the gastrointestinal tract to decipher the underlying mechanisms of how probiotics modulate immune responses to vaccines.

The effects of nano-curcumin supplementation on Th1/Th17 balance in migraine patients: A randomized controlled clinical trial

BackgroundThe present study was aimed to evaluate the nano-curcumin supplementation on Th1/Th17 balance by assessment of gene expression and serum level of interferon gamma (IFN-γ) and interleukin-17 (IL-17) in migraine patients. MethodsForty participants with episodic migraine were randomly allocated to receive 80 mg nano-curcumin (n = 20) or placebo (n = 20) in a randomized double-blind clinical trial for two months. The expression of IFN-γ and IL-17 from peripheral blood mononuclear cells and IFN-γ and IL-17 serum levels were measured, using a real-time PCR and ELISA methods, respectively. ResultsCompared to placebo group, two month nano-curcumin supplementation led to a significant reduction in serum levels and expression of IL-17 mRNA (P = 0.006 & 0.04, respectively), while there was no statistical difference regarding serum levels and expression of IFN-γ mRNA. ConclusionNano-curcumin supplementation in migraine patients led to a significant reduction in gene expression and plasma levels of IL-17 compared to control group.

Immunosuppressive effect of hispidulin in allergic contact dermatitis

BackgroundLong-term use of most immunosuppressants to treat allergic contact dermatitis (ACD) generates unavoidable severe side effects, warranting discovery or development of new immunosuppressants with good efficacy and low toxicity is urgently needed to treat this condition. Hispidulin, a flavonoid compound that can be delivered topically due to its favorable skin penetrability properties, has recently been reported to possess anti-inflammatory and immunosuppressive properties. However, no studies have investigated the effect of hispidulin on Th1 cell activities in an ACD setting.MethodsA contact hypersensitivity (CHS) mouse model was designed to simulate human ACD. The immunosuppressive effect of hispidulin was investigated via ear thickness, histologic changes (i.e., edema and spongiosis), and interferon-gamma (IFN-γ) gene expression in 1-fluoro-2,4-dinitrobenzene (DNFB)-sensitized mice. Cytotoxicity, total number of CD4+ T cells, and percentage of IFN-γ-producing CD4+ T cells were also investigated in vitro using isolated CD4+ T cells from murine spleens.ResultsTopically applied hispidulin effectively inhibited ear swelling (as measured by reduction in ear thickness), and reduced spongiosis, IFN-γ gene expression, and the number of infiltrated immune cells. The inhibitory effect of hispidulin was observed within 6 h after the challenge, and the observed effects were similar to those effectuated after dexamethasone administration. Hispidulin at a concentration up to 50 μM also suppressed IFN-γ-producing CD4+ T cells in a dose-dependent manner without inducing cell death, and without a change in total frequencies of CD4+ T cells among different concentration groups.ConclusionThe results of this study, therefore, suggest hispidulin as a novel compound for the treatment of ACD via the suppression of IFN-γ production in Th1 cells.

970P - Are immune checkpoint inhibitors a valid option for papillary renal cell carcinoma? Transcriptomic characterization of the immune infiltrate

BackgroundPapillary Renal Cell Carcinoma (pRCC) is the most common non-clear cell RCC (nccRCC). Pivotal studies evaluating immune checkpoint inhibitors mostly excluded nccRCC. PD-1/PD-L1 inhibitors exhibit limited activity in metastatic pRCC. The immune microenvironment in pRCC is unknown. MethodsIn silico, we studied the expression of cytotoxic lymphocyte infiltration (CYT), using a descriptive (by CIBERSORT) and specific quantitative approach, as well as the expression of inhibitors checkpoint immune markers (ICI), in 258 localized papillary renal cell tumors using RNA-seq data from The Cancer Genome Atlas (TCGA) as training set. Based on previous report, we selected 8 genes of interest (CD8a, CD8b, GZMA, PRF1, PD1, PDL1, PDL2 and CTLA4). An independent data set of 34 localized pRCC (gene expression) was used as a validation set. ResultsUsing a clustering method based on the expression level of 8 predefined genes of interest, we identified 3 groups, differentiated by CYT and ICI expression. In validation cohort, we observed similar clustering. Cluster 3, characterized by a CYT and ICI high expression, was significantly associated with increased population of TCD8, TCD4 helper, M1 macrophages and dendritic cells in CIBERSORT analysis. Additionally, these immune clusters were not associated with indels neo-antigen load but were significantly correlated with the MHC class I antigen presenting machinery expression (APM) (p=1.1.10-11) and interferon-gamma gene expression (p=1.6.10-13). ConclusionsWe characterized cytotoxic immune infiltration in pRCCs. Cluster 3 could correspond to a population of immunotherapy responders. Transcriptomic immune signature validation in pRCCs patients treated with immunotherapy is warranted. Legal entity responsible for the studyManon de Vries-Brilland. FundingHas not received any funding. DisclosureM. Gross-Goupil: Honoraria (institution), Advisory / Consultancy: Ipsen; Honoraria (institution), Advisory / Consultancy, Research grant / Funding (institution): Janssen; Honoraria (institution), Advisory / Consultancy: Astellas; Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: MSD; Research grant / Funding (institution): BMS; Research grant / Funding (institution): AstraZeneca; Research grant / Funding (institution), Travel / Accommodation / Expenses: Pfizer; Research grant / Funding (institution), Travel / Accommodation / Expenses: Roche; Travel / Accommodation / Expenses: Sanofi; Travel / Accommodation / Expenses: Amgen. A. Ravaud: Research grant / Funding (institution), Travel / Accommodation / Expenses, Officer / Board of Directors: Pfizer; Travel / Accommodation / Expenses, Officer / Board of Directors: BMS; Travel / Accommodation / Expenses, Officer / Board of Directors: AstraZeneca; Travel / Accommodation / Expenses, Officer / Board of Directors: Roche; Travel / Accommodation / Expenses, Officer / Board of Directors: MSD; Travel / Accommodation / Expenses, Officer / Board of Directors: Ipsen. B. Escudier: Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: BMS; Advisory / Consultancy, Research grant / Funding (institution), Travel / Accommodation / Expenses: Pfizer; Advisory / Consultancy, Research grant / Funding (institution): Novartis; Advisory / Consultancy, Research grant / Funding (self): Aveo; Advisory / Consultancy: Ipsen; Advisory / Consultancy: Roche; Advisory / Consultancy: EUSA. L. Albiges: Advisory / Consultancy: Pfizer; Advisory / Consultancy: Novartis; Advisory / Consultancy: BMS; Advisory / Consultancy: Ipsen; Advisory / Consultancy: Roche; Advisory / Consultancy: MSD; Advisory / Consultancy: AstraZeneca. All other authors have declared no conflicts of interest.

1310O - Adjuvant nivolumab (NIVO) versus ipilimumab (IPI) in resected stage III/IV melanoma: 3-year efficacy and biomarker results from the phase III CheckMate 238 trial

1310O - Adjuvant nivolumab (NIVO) versus ipilimumab (IPI) in resected stage III/IV melanoma: 3-year efficacy and biomarker results from the phase III CheckMate 238 trial