Expression Of Immune Response Genes Research Articles

The aryl hydrocarbon receptor (AHR) repressor limits expression of antimicrobial genes but not AHR-dependent genes in intestinal eosinophils.

Intestinal eosinophils express the aryl hydrocarbon receptor (AHR), an environmental sensor and ligand-activated transcription factor that responds to dietary or environmental ligands. AHR regulates tissue adaptation, survival, adhesion, and immune functions in intestinal eosinophils. The AHR repressor (AHRR) is itself induced by AHR and believed to limit AHR activity in a negative feedback loop. We analysed gene expression in intestinal eosinophils from WT and AHRR-KO mice and found that AHRR did not suppress most AHR-dependent genes. Instead, AHRR limited the expression of a distinct small set of genes involved in the innate immune response. These included S100 proteins, antimicrobial proteins and alpha-defensins. Using bone marrow-derived eosinophils we found that AHRR-KO eosinophils released more reactive oxygen species upon stimulation. This work shows that the paradigm of AHRR as a repressor of AHR transcriptional activity does not apply to intestinal eosinophils. Rather, AHRR limits the expression of innate immune response and antimicrobial genes, possibly to maintain an anti-inflammatory phenotype in eosinophils when exposed to microbial signals in the intestinal environment.

Investigation of post-immunization immune response gene expression kinetics in lymphoid tissues of White Leghorn and Indian native chicken

Understanding the basal expression kinetics of genes related to innate immunity aids effective genetic selection for immune response and how host genetics behaves with tolerance to Newcastle disease (ND) is obscure. It was hence aimed to investigate post-immunization mRNA expression kinetics of some immune response genes in White Leghorn, Aseel and Kadaknath chicken. Day-old chicks were vaccinated with ND RDF-1 strain with booster dose at 28-days of age at the experimental layer farm of ICAR-Central Avian Research Institute, Izatnagar (India) and lymphoid tissues were collected in RNAlater at 42-days of age. Total RNA was extracted and first strand cDNA was synthesized. Relative quantification of mRNA expression of IL1-β, IFN-γ, iNOS and TLR15 gene was assessed by qRT-PCR. The mRNA expressions significantly varied among the genes in spleen and thymus, and moderate expression of IFN-γ in all chicken genotypes suggested good protection against Newcastle disease virus. IL1-β-mRNA expressed at the highest level (P˂ 0.05) in spleen followed by thymus and bursa, whereas IFN-γ and iNOS expressed highly (P˂ 0.05) in thymus followed by spleen and bursa. In case of TLR15, the expression was significantly the highest in spleen followed by bursa and thymus. Kadaknath had the highest (P˂ 0.05) expression of IL1-β in thymus and that of iNOS in spleen, whereas Aseel showed the least (P > 0.05) expression than Kadaknath and White Leghorn. The study revealed the existence of wide variation in basal expression levels of immune response genes among different lymphoid tissues and lack of discernible differences in the expression between the chicken genotypes.

Select EZH2 inhibitors enhance viral mimicry effects of DNMT inhibition through a mechanism involving NFAT:AP-1 signaling.

DNA methyltransferase inhibitor (DNMTi) efficacy in solid tumors is limited. Colon cancer cells exposed to DNMTi accumulate lysine-27 trimethylation on histone H3 (H3K27me3). We propose this Enhancer of Zeste Homolog 2 (EZH2)-dependent repressive modification limits DNMTi efficacy. Here, we show that low-dose DNMTi treatment sensitizes colon cancer cells to select EZH2 inhibitors (EZH2is). Integrative epigenomic analysis reveals that DNMTi-induced H3K27me3 accumulates at genomic regions poised with EZH2. Notably, combined EZH2i and DNMTi alters the epigenomic landscape to transcriptionally up-regulate the calcium-induced nuclear factor of activated T cells (NFAT):activating protein 1 (AP-1) signaling pathway. Blocking this pathway limits transcriptional activating effects of these drugs, including transposable element and innate immune response gene expression involved in viral defense. Analysis of primary human colon cancer specimens reveals positive correlations between DNMTi-, innate immune response-, and calcium signaling-associated transcription profiles. Collectively, we show that compensatory EZH2 activity limits DNMTi efficacy in colon cancer and link NFAT:AP-1 signaling to epigenetic therapy-induced viral mimicry.

Dietary supplementation of micelle silymarin enhances the antioxidant status, innate immunity, growth performance, resistance against Vibrio parahaemolyticus infection, and gut morphology in Pacific white shrimp (Litopenaeus vannamei)

Silymarin, a flavonoid compound derived from the milk thistle plant (Silybum marianum) is well-known for its hepatoprotective, antioxidant, immunomodulatory, and anti-inflammatory properties. Therefore, this study sought to evaluate the effect of silymarin and micelle silymarin (MS) supplementation on Pacific white shrimp (Litopenaeus vannamei). The experimental diets were formulated to include MS at levels of 0.25, 0.5, 1.0, 2.0, and 4.0 g/kg or silymarin at levels of 1.0 and 2.0 g/kg in the basal diet (Con, M0.25, M0.5, M1, M2, M4, S1 and S2, respectively). Juvenile shrimp (0.98 g) were fed with experimental diets (four replicates) for 42 days. The growth metrics were significantly (P < 0.05) increased in M0.25, M0.5, and M1 groups compared to the Con group. Feed conversion ratio decreased (P < 0.05) in M0.5, M1, and M2 groups, whereas the protein efficiency ratio increased (P < 0.05) in M0.5 and M1 groups compared to the Con group. Phenoloxidase, lysozyme, glutathione peroxidase, and superoxide dismutase activities were enhanced (P < 0.05), with the highest levels observed in the M0.5 group. Malondialdehyde level decreased (P < 0.05) in M1 and M2 groups compared to the Con group. Total protein increased (P < 0.05) in M0.25, M0.5 and S1 groups compared to the Con group. AST levels were significantly reduced (P < 0.05) in shrimp fed M0.5, M1, M2, M4, and S2 groups, and ALT levels decreased (P < 0.05) in M1 and S2 groups compared to Con group. Wall thickness was significantly higher (P < 0.05) in the M0.5 group, and villi length increased (P < 0.05) in M0.25, M0.5, and S1 groups. The resistance against Vibrio parahaemolyticus increased; however, no significant differences in cumulative survival were observed (P ˃ 0.05). Significantly higher (P < 0.05) proPO levels were observed in M0.5, M1, M2, M4, and S2 groups, whereas higher (P < 0.05) TLR-3 levels were observed in M1 and M2 groups compared to the Con group. TNF-α expression decreased (P < 0.05) in all other dietary treatments compared to the Con group. The findings suggest that dietary silymarin and MS can enhance growth, immune response, antioxidant capacity, gut morphology, and immune gene expression, while reducing lipid peroxidation and inflammation in Pacific white shrimp. The shrimp performance shows no significant difference between silymarin and MS groups at 1% and 2% levels. The optimal suggested inclusion level of MS in shrimp feed is 0.5–1.0 g/kg.

Steroid immune responsive gene regulation in Mycobacterium tuberculosis infection in vitro

Tuberculosis (TB) is an infectious disease displaying a multifactorial pathology. The immunomodulatory role attributed to steroid hormones, such as vitamin D3 (VD3) and 17β-estradiol (E2), highlighted the importance of these hormones against Mycobacterium tuberculosis (Mtb) infection. In order to understand their influence upon gene expression of immune and inflammatory responsive genes against Mtb we tested it in vitro using peripheral blood mononuclear cells (PBMCs). Cells were pretreated with VD3 (50 ng/mL) or E2 (100 nM/mL) and co-cultured with H37Rv Mtb or stimulated with lipopolysaccharide from Escherichia coli (LPS). After 24 h and 72 h of co-culture the Mtb viability in macrophages test was performed, as well the total RNA isolation for gene expression analysis by RT-qPCR of the following target genes: NLRP3, DC-SIGN, IL-1β, and IL-10. We also measured IL-10, TNF, IFN-γ, IL-4, IL-6, and IL-2 supernatant levels. As the main results, we found that VD3 and E2 downregulated the expression of inflammatory genes NLRP3, IL-1β, and IL-10 expression in Mtb co-cultured cells. Finally, VD3 treatment increased the release of the cytokine IFN-γ in Mtb-infected cells, while E2 treatment inhibited the release of IL-10, TNF, IFN-γ, and IL-6. Therefore, we report an immunogenetic influence of VD3 and E2 upon Mtb co-culture.

Abstract TP33: Knockout of Kruppel-Like Factor 4 in Brain Endothelial Cells Accelerates Vascular and Neuronal Degeneration During Aging

As we get older, various aspects of our bodies start to decline, including brain function. The mechanisms behind the decline in brain function due to aging are still not well understood. The blood-brain barrier (BBB) is crucial for maintaining brain health and homeostasis by controlling both passive and active exchange of materials between the blood and brain. This barrier is predominantly composed of endothelial cells, which deteriorate in both aging and neurodegenerative disorders. Endothelial cells are also highly enriched in the Kruppel-like factor 4 (KLF4) transcription factor, which is reduced in expression as we age. In this study, we report that endothelial-specific KLF4 KO mice demonstrate accelerated brain aging, including accelerated BBB deterioration, neurodegeneration, and cognitive decline. Single-cell RNA sequencing from brain endothelial cells of endothelial-specific KLF4 KO mice demonstrate aberrantly high expression of adaptive and innate immune response genes, and live animal brain imaging across the lifespan reveals accelerated aging-related BBB deterioration, decreased blood flow, and impaired neurovascular coupling. Endothelial-specific KLF4 KO mice show increased neuroinflammation and neutrophil infiltration. Our data suggest that diminished endothelial cell KLF signaling with age could be a novel therapeutic target to preserve health and function of the aging brain.

Presence of Epstein–Barr virus (EBV) antigens detected by sensitive methods has no influence on local immune environment in diffuse large B cell lymphoma

EBV+ diffuse large B cell lymphoma (DLBCL) not otherwise specified (NOS) is a new entity confirmed by the World Health Organization (WHO) in 2017. In this new entity, the virus may contribute to a tolerogenic microenvironment. Traces of the virus have been described in DLBCL with more sensitive methods, in cases that were originally diagnosed as negative. The aim of this study was to analyze the expression of immune response genes in the tumor microenvironment to disclose the role of the virus and its traces in DLBCL. In 48 DLBCL cases, the expression of immune response genes and the presence of molecules that induce tolerance, such as TIM3, LAG3 and PDL1 by immunohistochemistry (IHC), were studied. To broaden the study of the microenvironment, tumor-associated macrophages (TMAs) were also explored. No significant differences were observed in the expression of immune response genes in the EBV+ DLBCL and those cases that were EBV− DLBCL but that exhibited viral traces, assessed by ViewRNA assay. Only the EBV+ DLBCL cases displayed a significantly higher increase in the expression of CD8 and cytotoxic T cells detected by gene expression analysis, and of PDL1 in tumor cells and in the expression of CD68 in the tumor microenvironment detected by IHC, not observed in those cases with viral traces. The increase in CD8 and cytotoxic T cells, PDL1 and CD68 markers only in EBV+ DLBCL may indicate that traces of viral infection might not have influence in immune response markers.

Increased abundance of potentially pathogenic Vibrio and a marine heatwave co-occur with a Pacific Oyster summer mortality event

The Pacific Oyster (Crassostrea gigas) is an important aquaculture species, that has been globally affected by “summer mortality” outbreaks. The causes of summer mortality are often unclear, but several contributing factors have been implicated. To better understand the factors contributing to C. gigas summer mortality, we performed a 24 week experiment in tanks filled by a continuous flow of unfiltered surface seawater during a summer mortality high-risk period. Throughout this period, we measured a suite of environmental parameters, with weekly sampling to characterise the bacterial community composition, the abundance and diversity of the Vibrio genus, and the expression of oyster immune response genes. We observed three clear phases of oyster mortality: Phase 1 (day 0 to 75) was characterised by sporadic and low levels (mean < 1% per week) of oyster mortality; Phase 2 (day 82 to 119) involved constant levels of ongoing mortality (mean < 10% per week); and Phase 3 (day 125 to 161) involved a sharp increase in mortality (mean > 10% per week) reaching up to weekly mortality of 51%. The shift between phase 2 and 3 coincided with heavy rainfall and a marine heatwave (MHW) event at our study site, which lasted 13 days, when the average water temperature was elevated by 0.8 °C when compared to the weeks prior and following the MHW, and a maximum water temperature of 27.7 °C was reached. Oyster mortality was positively correlated with the occurrence of the MHW as well as bacterial and Vibrio abundance, and negatively correlated with seawater salinity that decreased from a salinity of 34.6 to 31.4 psu following rainfall events. In addition, the relative abundance of several Vibrio species, including V. campbellii, V. rotiferianus and V. owensii displayed positive correlations to oyster mortality. Shifts in the oyster-associated bacterial community and oyster immunity gene expression profiles occurred in parallel to increased mortality levels, with genes involved in the regulation of antimicrobial peptides (IkB1 and NF-kB p105) significantly over-expressed during high mortality. Overall, we observed a C. gigas mortality event that coincided with a marine heatwave, rainfall event and a shift in bacterial community composition. Our analyses provides further evidence for the putative role of Vibrio in summer mortality events.

Host and Microbe Blood Metagenomics Reveals Key Pathways Characterizing Critical Illness Phenotypes.

Rationale: Two molecular phenotypes of sepsis and acute respiratory distress syndrome, termed hyperinflammatory and hypoinflammatory, have been consistently identified by latent class analysis in numerous cohorts, with widely divergent clinical outcomes and differential responses to some treatments; however, the key biological differences between these phenotypes remain poorly understood.Objectives: We used host and microbe metagenomic sequencing data from blood to deepen our understanding of biological differences between latent class analysis-derived phenotypes and to assess concordance between the latent class analysis-derived phenotypes and phenotypes reported by other investigative groups (e.g., Sepsis Response Signature [SRS1-2], molecular diagnosis and risk stratification of sepsis [MARS1-4], reactive and uninflamed).Methods: We analyzed data from 113 patients with hypoinflammatory sepsis and 76 patients with hyperinflammatory sepsis enrolled in a two-hospital prospective cohort study. Molecular phenotypes had been previously assigned using latent class analysis.Measurements and Main Results: The hyperinflammatory and hypoinflammatory phenotypes of sepsis had distinct gene expression signatures, with 5,755 genes (31%) differentially expressed. The hyperinflammatory phenotype was associated with elevated expression of innate immune response genes, whereas the hypoinflammatory phenotype was associated with elevated expression of adaptive immune response genes and, notably, T cell response genes. Plasma metagenomic analysis identified differences in prevalence of bacteremia, bacterial DNA abundance, and composition between the phenotypes, with an increased presence and abundance of Enterobacteriaceae in the hyperinflammatory phenotype. Significant overlap was observed between these phenotypes and previously identified transcriptional subtypes of acute respiratory distress syndrome (reactive and uninflamed) and sepsis (SRS1-2). Analysis of data from the VANISH trial indicated that corticosteroids might have a detrimental effect in patients with the hypoinflammatory phenotype.Conclusions: The hyperinflammatory and hypoinflammatory phenotypes have distinct transcriptional and metagenomic features that could be leveraged for precision treatment strategies.

Differential susceptibility of human microglia HMC3 cells and brain microvascular endothelial HBEC-5i cells to Mayaro and Una virus infection

Mayaro (MAYV) and Una (UNAV) are emerging alphaviruses circulating in the Americas. Earlier reports have revealed that MAYV infects different human cell lines, including synovial and dermal fibroblasts, chondrocytes, osteoblasts, astrocytes and pericytes, as well as neural progenitor cells. In this study we evaluated the susceptibility of immortalized human microglia HMC3 cells and brain microvascular endothelial HBEC-5i cells to MAYV and UNAV infection. Cytopathic effects, cell viability, viral progeny yields, and the presence of E1 and nsP1 proteins in HMC3 and HBEC-5i cells infected with several MAYV or UNAV strains were assessed using an inverted microscope, MTT assay, plaque-forming assays, and immunofluorescence or Western blot, respectively. Finally, the expression of immune response genes was analyzed using RT-qPCR. MAYV and UNAV demonstrated strong cytopathic effects and significantly reduced cell viability in HMC3 cells. Moreover, the HMC3 cells were efficiently infected regardless of the virus strain tested, and E1 and nsP1 viral proteins were detected. In contrast, only MAYV appeared to infect HBEC-5i cells, and minimal effects on cell morphology or viability were observed. Furthermore, the MAYV titer and viral protein levels were substantially lower in the infected HBEC-5i cells when compared to those of the infected microglia cells. Finally, unlike UNAV, MAYV elicited a strong expression of specific interferon-stimulated genes in microglia cells, along with pro-inflammatory cytokines implicated in the immune response. Collectively, these findings demonstrate that MAYV and UNAV are capable of infecting relevant human brain cells.

Effects of Antrodia cinnamomea solid culture mycelium by-products on growth performance and immune response in weaning black piglets.

This study evaluated the effects of supplementation with Antrodia cinnamomea mycelium by-product (ACBP) on growth performance and immune response in weaning piglets. Total available content and antioxidant capacity of ACBP were determined. Ninety-six black pigs were randomly distributed to 24 pens. Study compared four groups which were supplemented with ACBP at 0%, 2.5%, 5%, or 10% for 6 weeks after weaning at 4 weeks. Results showed that ACBP on total phenolic, total flavonoid, and total triterpenoids contents were 13.68 mg GAE/g DW, 1.67 μg QE/g DW, and 15.6 mg/g, respectively. Weaning piglets fed 2.5% ACBP showed a significant decreased body weight gain compared with those supplemented with 5% ACBP, 10% ACBP, and control groups. Results showed that all ACBP groups increased the villi height of jejunum significantly. Incidence of diarrhea in 11 weeks with supplementation with 5% and 10% ACBP diets were lower than in control group. The 10% ACBP group showed significantly lower expression of immune response genes (IL-1β, IL-6, IL-8, TNF-α, and IFN-γ) than the 2.5% and 5% ACBP groups. Based on results, dietary supplementation with 10% ACBP did not significantly affect body weight but could decrease piglet diarrhea condition and expression of IL-1β and IL-6 genes.

Interactions of the Immune System with Human Kidney Organoids.

Kidney organoids are an innovative tool in transplantation research. The aim of the present study was to investigate whether kidney organoids are susceptible for allo-immune attack and whether they can be used as a model to study allo-immunity in kidney transplantation. Human induced pluripotent stem cell-derived kidney organoids were co-cultured with human peripheral blood mononuclear cells (PBMC), which resulted in invasion of allogeneic T-cells around nephron structures and macrophages in the stromal cell compartment of the organoids. This process was associated with the induction of fibrosis. Subcutaneous implantation of kidney organoids in immune-deficient mice followed by adoptive transfer of human PBMC led to the invasion of diverse T-cell subsets. Single cell transcriptomic analysis revealed that stromal cells in the organoids upregulated expression of immune response genes upon immune cell invasion. Moreover, immune regulatory PD-L1 protein was elevated in epithelial cells while genes related to nephron differentiation and function were downregulated. This study characterized the interaction between immune cells and kidney organoids, which will advance the use of kidney organoids for transplantation research.

Effect of Lincomycin and Butyrate Glycerides Supplementation on Performance, Blood Biochemical Constituents, Immune Response and Nutrient Absorption Related Gene Expression in Broilers

Abstract Over 35 days feeding trial, the effects of lincomycin (LM) and butyrate glycerides (BG) supplementation on the growth performance, blood biochemical changes, immune response, and nutrient absorption related gene expression in broiler chickens were evaluated. A total of 480 one- day-old broiler chicks (Ross 308) were allotted into 6 groups (8 replicates/group with 10 birds/replicate). Experimental groups were arranged as follows; control group (A) fed the basal diet (BD); B, C, and D groups were fed the BD supplemented with 350, 500 and 1000 g BG/ton of feed respectively. While group E was fed the BD with 350 + 500 + 1000 g BG/ton of feed during starter, grower, and finisher periods, respectively, and finally group F which received lincomycin (LM) antibiotic (100 g/ton) in overall periods. Broilers that received BG in group E (birds supplemented with BG at 350 + 500 + 1000 g/ton feed) and LM (F) supplements both gained more body weight and weight growth (P&lt;0.05). Lower feed intake and improved fcr were observed in all the experimental groups that received either BG or lm compared to control birds (P&lt;0.05). Lower serum total cholesterol, triglycerides, and glutamic oxaloacetic transaminase (GOT) (P&lt;0.05) were recorded in the lm or BG supplemented birds compared to the control. Higher immune response represented by enhanced antibody titer against H9N1, and newcastle disease (ND) and lower hepatic malondialdehyde (MDA) was obtained in groups D, E and F (P&lt;0.05) compared to the other groups. Higher gene expression of GSH-Px, Glut 2, PepT1 and FAS was obtained in groups D, E and F (P&lt;0.05) compared to the control. Supplemental BG or LM increased the unsaturated fatty acids (oleic, linoleic and linolenic acids) while lowered the palmitic acid contents in breast muscle (P&lt;0.05) compared to control. In conclusion, supplementing broilers’ diets with BG or lincomycin improved their growth performance, immune response and muscle fatty acid profile as well as modulatory role in lipid metabolism. Supplementing starter, grower, and finisher broiler diets with BG at 350 + 500 + 1000 g/ton each may be a useful substitute for lincomycin.

An Investigation on the Effects of Dietary Vitamin E on Juvenile Sea Urchin (Strongylocentrotus intermedius): Growth, Intestinal Microbiota, Immune Response, and Related Gene Expression.

A 90 d feeding experiment was conducted to investigate the effects of vitamin E (VE) on growth, intestinal microbiota, immune response, and related gene expression of juvenile sea urchin (Strongylocentrotus intermedius). Six dry feeds were made to contain graded levels of VE (78, 105, 152, 235, 302, and 390 mg/kg); these were named E78, E105, E152, E235, E302, and E390, respectively. Dry feed E50 and fresh kelp (HD) were used as the control diets. There were six replicates of cages in each dietary group, and each cage held 20 sea urchins with an initial body weight of approximately 1.50 g. Results exhibited that weight gain rate and gonadosomatic index (GSI) of the sea urchins were not significantly affected by dietary VE ranging from 78 to 390 mg/kg. Sea urchins in the dry feed groups showed poorer growth performance, but significantly higher GSI than those in the fresh kelp groups. The pepsin and lipase activities were not significantly promoted by low or moderate VE, but were inhibited by a high level of VE (302-390 mg/kg), while amylase and cellulase activities were significantly increased by low or moderate VE, with the highest values observed in the E105 and E235 groups, respectively. VE addition at a low dosage (105-152 mg/kg) showed inhibitory effects on immune and antioxidant enzyme activities and expression of inflammation-related genes, but showed no beneficial effects at moderate or high dosage (235-390 mg/kg), while a moderate or relatively higher level of VE (235-302 mg/kg) significantly increased the expression of several immune-related genes. The relative abundance of Proteobacteria, Actinobacteria, Ruegeria, and Maliponia in the intestine of the sea urchins increased with the increase in VE in the dry feeds. On the contrary, the relative abundance of the Firmicutes, Bacteroidetes, Escherichia-Shigella, Bacteroides, and Clostridium sensu stricto 1 gradually decreased as VE content increased. These results indicated that a moderate level of VE (172.5-262.4) can achieve ideal digestive enzyme activities and growth performance, but a relatively higher level of VE (235-302 mg/kg) was beneficial for maintaining the immune and antioxidant capacity of juvenile S. intermedius by regulating the expression of inflammation- and immune-related genes and abundance of some bacteria to a healthy state.

A combination of burn wound injury and Pseudomonas infection elicits unique gene expression that enhances bacterial pathogenicity.

Burns are a leading cause of morbidity and mortality worldwide with the most common cause of death resulting from sepsis, often from Pseudomonas aeruginosa. We previously reported that a non-lethal flame burn induced an altered host immune response. Using this model, gene expression in both the murine host and P. aeruginosa was measured using a NanoString custom probe panel. We observed differing patterns of gene expression in both host and P. aeruginosa in the skin, blood, liver, and spleen of mice that were burned and/or infected, compared to mice that were neither burned nor infected (i.e., Sham). In mice that were both burned and infected (B/I), we observed changes in gene expression in both the host and P. aeruginosa that were distinct from all other treatment conditions. These data suggest that the combination of the burned state and superimposed infection affects both host and pathogen gene expression to increase infection propensity. Gene transcription significantly changed from 6 to 24 h post-B/I in each tissue. Finally, inhibiting IL-10 signaling or co-administering arginine at the time of P. aeruginosa infection prolonged or restored survival in an otherwise 100% fatal burn and infection model. These findings suggest that disease states such as burns may differentially alter innate immune response gene expression in both a host- and pathogen-specific manner.IMPORTANCEThe interaction between an underlying disease process and a specific pathogen may lead to the unique expression of genes that affect bacterial pathogenesis. These genes may not be observed during infection in the absence of, or with a different underlying process or infection during the underlying process with a different pathogen. To test this hypothesis, we used Nanostring technology to compare gene transcription in a murine-burned wound infected with P. aeruginosa. The Nanostring probeset allowed the simultaneous direct comparison of immune response gene expression in both multiple host tissues and P. aeruginosa in conditions of burn alone, infection alone, and burn with infection. While RNA-Seq is used to discover novel transcripts, NanoString could be a technique to monitor specific changes in transcriptomes between samples and bypass the additional adjustments for multispecies sample processing or the need for the additional steps of alignment and assembly required for RNASeq. Using Nanostring, we identified arginine and IL-10 as important contributors to the lethal outcome of burned mice infected with P. aeruginosa. While other examples of altered gene transcription are in the literature, our study suggests that a more systematic comparison of gene expression in various underlying diseases during infection with specific bacterial pathogens may lead to the identification of unique host-pathogen interactions and result in more precise therapeutic interventions.

Systematic evaluation of AML-associated antigens identifies anti-U5 SNRNP200 therapeutic antibodies for the treatment of acute myeloid leukemia

Despite recent advances in the treatment of acute myeloid leukemia (AML), there has been limited success in targeting surface antigens in AML, in part due to shared expression across malignant and normal cells. Here, high-density immunophenotyping of AML coupled with proteogenomics identified unique expression of a variety of antigens, including the RNA helicase U5 snRNP200, on the surface of AML cells but not on normal hematopoietic precursors and skewed Fc receptor distribution in the AML immune microenvironment. Cell membrane localization of U5 snRNP200 was linked to surface expression of the Fcγ receptor IIIA (FcγIIIA, also known as CD32A) and correlated with expression of interferon-regulated immune response genes. Anti-U5 snRNP200 antibodies engaging activating Fcγ receptors were efficacious across immunocompetent AML models and were augmented by combination with azacitidine. These data provide a roadmap of AML-associated antigens with Fc receptor distribution in AML and highlight the potential for targeting the AML cell surface using Fc-optimized therapeutics.

Microbial mechanisms of regulation of the activity of nuclear transcription factor NF-kB1

Nuclear transcription factor NF-kB1 is a regulator of the expression of immune response genes, apoptosis, inflammatory reactions and cell proliferation. Microorganisms are able to change the activity of the nuclear transcription factor, which allows them to modulate the immune response of the macroorganism and/or have a pathogenic effect on organs and tissues. The aim of the study is to systematize data on microbial molecular factors regulating the transcription nuclear factor NFkB-1. The review describes microbial molecules, the mechanisms of their action on the nuclear transcription factor NF-kB1. The A52R protein of the smallpox vaccine virus acts as a homologue of MyD88, the Yersinia YopP/J protein acetylates the IKKß subunit, thereby inhibiting kB kinase (IKK) and preventing the activation of the nuclear factor NFkB. The EBNA1 protein of the Epstein-Barr virus is a "stop signal" for the proteasome, so blocking proteolysis disrupts the presentation of virus antigens on MHC I and the overall immune response. The OspG protein of microorganisms of the genus Shigella prevents ubiquitination, and also inhibits the degradation of phosphorylated IKKß, which disrupts the activation of NFkB-1. Enteropathogenic Escherichia coli (EPEC) produces three effectors to inhibit the function of NFkB. Two of these effectors, NleB and NleE block signaling at the level of TGF-beta-activated kinase 1 or IKKß kinase, the third NleH protein binds to the RPS3 (ribosomal protein S3) cofactor of the NFkB factor and suppresses the transcription of many genes. The components of bacterial cells – lipopolysaccharides of the cell walls of gram-negative bacteria, representatives of the Enterobacteriaceae family, participate in modulating the activity of the transcription factor. This polymer affects the presentation genes of MHC class I and II antigens, the death receptor, IL-6 signaling, IFN-γ signaling and the P38 signaling pathway. Conclusion. Protein and non-protein substances of microorganisms, components of bacterial cell structures are molecules that have a multidirectional effect on the activity of proteasomes that regulate inflammatory and immune responses in the macroorganism.

Impact of dietary piperine on growth performance, immune response, antioxidant status, and immune-related gene expression in pathogen-aggravated Cyprinus carpio

This study investigated the effects of dietary piperine (PIP) on growth performance, digestive enzymes, serum biochemical parameters, antioxidant and immune responses, and gene expression in Cyprinus carpio challenged with Aeromonas hydrophila. Six diets were prepared with PIP doses of 0, 0.5, 1.0, 2.0, 3.0, and 4.0 g/kg, corresponding with the control, PR50, PR100, PR200, PR300, and PR400, respectively. Fish were challenged with Aeromonas hydrophila after 8 weeks of feeding with the respective diets. Weight gain (PWG) and specific growth rate (SGR) were significantly enhanced, whereas feed conversion ratio (FCR) was lowered in PR200. The cumulative post-challenge survival was improved to 68.43% in the PR200 group compared with 28.08% in the control. Serum total protein and albumin levels were significantly enhanced in the PR200 group compared to the control. However, dietary PIP up to 3 g/kg had no significant effect on serum glucose, cortisol, aspartate aminotransferase, or alkaline phosphatase activities; however, the alanine aminotransferase level was lower (P < 0.05) in the PR200 group than in the control. Intestinal amylase, lipase, and protease activities increased in PR300, and intestinal amylase and lipase increased in the PR100 group (P < 0.05). The serum immunological indices (lysozyme, alternative complement pathway, phagocytic activity, and respiratory burst activity) were higher (P < 0.05) in the PR200 group than in the control group. Serum superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities were significantly intensified in PR200–PR300 than in the control group, with the highest activity observed in the PR200 group. Malondialdehyde was significantly lower in the PR200 group than in the control group. Furthermore, SOD, CAT, and Nrf2 expression was strongly upregulated in the liver tissue of the PR200 and PR300 groups compared to that in the control. The transcript levels of pro-inflammatory cytokines viz. IL-1β and TNF-α were significantly upregulated in the kidneys of the PR100 and PR200 post-challenged. In contrast, the anti-inflammatory cytokine IL-10 was significantly downregulated in the kidneys of PR200. The expression of HSP70 was upregulated only in the PR400. Quadratic regression analysis showed that the optimal dietary PIP level was estimated as 2.07−2.13 g/kg to maximize growth performance. Overall, these results indicate that dietary PIP at an appropriate level can improve immunity, cytokine gene expression, and disease resistance in C. carpio.

Periodontitis-resistant and -susceptible matriline regulation of gingival transcriptome in nonhuman primates.

This report identified if gingival gene expression transcriptomes demonstrated unique profiles that discriminated periodontitis-susceptible (PDS) and periodontitis-resistant (PDR) animals in health and disease. Nonhuman primates generally organize their social groups based upon matriline origin. We have used a multi-generational colony of rhesus macaques to identify matrilines presenting with significant differences in periodontitis (e.g., earlier age onset, greater prevalence, and severity). Animals from 12 to 23 years of age (n = 17; 8 - PDR, 9 - PDS) were entered into a ligature-induced periodontitis trial. Gingival biopsies were taken at baseline and 0.5, 1, 3, and 5 months post-ligation, and microarray analysis was used to quantify gene expression in samples at each time point. Over 1000 genes showed significant (p < .01) differences in the PDR versus PDS animals at baseline. The frequency of differences generally decreased during the disease process, and increased with resolution (i.e., 5 months). A nearly 2:1 ratio of elevated gene levels was noted in baseline PDR samples that included up-regulated MMPs, Fc receptors, chemokines, interleukins, and innate immune receptors, and down-regulated genes particularly related to epithelial biology. Most dramatically, there was a skewed differential expression of adaptive immune response genes in the PDR and epithelial cell structure/function genes in PDS samples. The results demonstrate substantive differences in gingival tissue response capacity/programming in PDR and PDS samples that may contribute to the differences in clinical outcomes related to the heritability of disease risk through matrilines.

SPOP targets the immune transcription factor IRF1 for proteasomal degradation.

Adaptation of the functional proteome is essential to counter pathogens during infection, yet precisely timed degradation of these response proteins after pathogen clearance is likewise key to preventing autoimmunity. Interferon regulatory factor 1 (IRF1) plays an essential role as a transcription factor in driving the expression of immune response genes during infection. The striking difference in functional output with other IRFs is that IRF1 also drives the expression of various cell cycle inhibiting factors, making it an important tumor suppressor. Thus, it is critical to regulate the abundance of IRF1 to achieve a 'Goldilocks' zone in which there is sufficient IRF1 to prevent tumorigenesis, yet not too much which could drive excessive immune activation. Using genetic screening, we identified the E3 ligase receptor speckle type BTB/POZ protein (SPOP) to mediate IRF1 proteasomal turnover in human and mouse cells. We identified S/T-rich degrons in IRF1 required for its SPOP MATH domain-dependent turnover. In the absence of SPOP, elevated IRF1 protein levels functionally increased IRF1-dependent cellular responses, underpinning the biological significance of SPOP in curtailing IRF1 protein abundance.