How obesity and metabolic syndrome trigger non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC) remains elusive. In this issue, Gomes and colleagues describe that nutrient surplus induces hepatic URI expression, triggering genotoxicity and IL17A expression, thus leading to insulin resistance, NASH, and HCC. IL17A signaling blockers might become a readily translatable therapy.

Mammalian pregnancy evolved in the therian stem lineage, that is, before the common ancestor of marsupials and eutherian (placental) mammals. Ancestral therian pregnancy likely involved a brief phase of attachment between the fetal and maternal tissues followed by parturition-similar to the situation in most marsupials including the opossum. In all eutherians, however, embryo attachment is followed by implantation, allowing for a stable fetal-maternal interface and an extended gestation. Embryo attachment induces an attachment reaction in the uterus that is homologous to an inflammatory response. Here, we elucidate the evolutionary mechanism by which the ancestral inflammatory response was transformed into embryo implantation in the eutherian lineage. We performed a comparative uterine transcriptomic and immunohistochemical study of three eutherians, armadillo (Dasypus novemcinctus), hyrax (Procavia capensis), and rabbit (Oryctolagus cuniculus); and one marsupial, opossum (Monodelphis domestica). Our results suggest that in the eutherian lineage, the ancestral inflammatory response was domesticated by suppressing one of its modules detrimental to pregnancy, viz. neutrophil recruitment by cytokine IL17A. Further, we propose that this suppression was mediated by decidual stromal cells, a novel cell type in eutherian mammals. We tested a prediction of this model in vitro, and showed that decidual stromal cells can suppress the production of IL17A from helper T cells. Together, these results provide a mechanistic understanding of early stages in the evolution of eutherian pregnancy.

Eomesodermin (Eomes), a transcription factor, could suppress the Th17 cell differentiation and proliferation through directly binding to the promoter zone of the Rorc and Il17a gene, meanwhile the expression of Eomes is suppressed when c-Jun directly binds to its promoter zone. Ginkgolide K (1,10-dihydroxy-3,14-didehydroginkgolide, GK) is a diterpene lactone isolated from the leaves of Ginkgo biloba. A previous study indicated that GK could decrease the level of phospho JNK (c-Jun N-terminal kinase). Here, we reported the therapeutic potential of Ginkgolide K (GK) treatment to ameliorate experimental autoimmune encephalomyelitis (EAE) disease progression. Methods: EAE was induced in both wildtype and CD4-Eomes conditional knockout mice. GK was injected intraperitoneally. Disease severity, inflammation, and tissue damage were assessed by clinical evaluation, flow cytometry of mononuclear cells (MNCs), and histopathological evaluation. Dual-luciferase reporter assays were performed to measure Eomes transcription activity in vitro. The potency of GK (IC₅₀) was determined using JNK1 Kinase Enzyme System. Results: We revealed that GK could ameliorate EAE disease progression by the inhibition of the Th17 cells. Further mechanism studies demonstrated that the level of phospho JNK was decreased and the level of Eomes in CD4⁺T cells was dramatically increased. This therapeutic effect of GK was almost completely interrupted in CD4-Eomes conditional knockout mice. Conclusions: These results provided the therapeutic potential of GK treatment in EAE, and further suggested that Eomes expression in CD4⁺T cells might be essential in this process.

Keywords: Multiple sclerosis· Transcription factors· EAE· Therapeutic target.

Campylobacter jejuni is among the leading causes of bacterial foodborne illness. Poultry is the major reservoir and source of human campylobacteriosis. Currently, there is no effective and practical method to decrease C. jejuni colonization in chickens or to reduce human infections. Additionally, antibiotic-resistant infections pose a serious public health concern; therefore, antibiotic-alternative approaches are needed to reduce transmission of C. jejuni including resistant bacteria from chickens to humans. Here, we evaluated the effect of E. coli Nissle 1917 (EcN) on innate responses of polarized HT-29 cells and consequently on C. jejuni 81176 infections in HT-29 cells. Pre-treatment of HT-29 cells with EcN for 4 h had a significant effect on the invasion of different C. jejuni strains (2 h post-infection) (P < .05) and no intracellular C. jejuni (24 h post-infection) were recovered. To further understand how EcN mediates its impact on C. jejuni's survival inside the cells, we used Human Antibacterial RT² Profiler^TM PCR arrays to profile gene expression in HT-29 cells after treatment with EcN with or without C. jejuni 81-176 infection. Our results suggest that pre-treatment of the HT-29 cells with EcN induced the anti-inflammatory cytokines and activated the anti-apoptotic Akt signaling which likely to protect the cells against the proinflammatory and apoptosis responses induced by C. jejuni. EcN also positively affected the expression of genes involved in cellular maintenance, growth, development, and proliferation. Further, EcN modulated the expression of genes involved in protective innate immunity, such as TLRs, ERK1/2, p38 MAPK, Ap1, JNK, IL1B, IL17A, and NF-κB signaling.

Keywords: Campylobacter; E. coli Nissle 1917; HT-29 cells; Probiotic; innate immune response; intracellular survival; invasion.

Th17 lymphocytes, that produce IL17A, IL17F, and IL22, play a crucial role during the immune response against Mycobacterium tuberculosis (Mtb) infection. Whereas, the contribution of IL17A in immunity to tuberculosis is usually accepted, the role of IL17F has been scarcely studied so far. The aim of this work was to evaluate the existence of a potential association of the non-synonymous variant rs763780 SNP of the IL17F gene with human tuberculosis. Accordingly, by comparing healthy donors (HD) and tuberculosis patients (TB) populations we demonstrated an association between the C allele of the SNP and the susceptibility to tuberculosis disease in Argentina. Furthermore, we found that peripheral blood mononuclear cells (PBMCs) from individuals with a more effective immune response against Mtb secreted the highest levels of IL17F when stimulated with a lysate of Mtb (Mtb-Ag). Besides, we evidenced that Mtb-Ag-stimulated PBMCs from HD carrying the C variant of the SNP displayed the lowest IFNG secretion, proliferation index, and SLAM expression as compared to TT carriers. Moreover, Mtb-Ag-stimulated PBMCs from TB carrying the C allele produced the lowest levels of IFNG, the highest level of IL17A, and the minimum proliferation indexes as compared to TT TB, suggesting a relationship between the C allele and tuberculosis severity. In fact, TB carrying the C allele presented a more severe disease, with the highest bacilli burden in sputum. Together, our findings identify the IL17F rs763780 SNP as a biomarker of tuberculosis susceptibility and advanced disease severity in Argentina, suggesting that IL17F could be a critical cytokine in tuberculosis immunity.

Autosomal dominant hyper-IgE syndrome caused by dominant-negative loss-of-function mutations in signal transducer and activator of transcription factor 3 (STAT3) (STAT3-HIES) is a rare primary immunodeficiency with multisystem pathology. The quality of life in patients with STAT3-HIES is determined by not only the progressive, life-limiting pulmonary disease, but also significant skin disease including recurrent infections and abscesses requiring surgery. Our early report indicated that hematopoietic stem cell transplantation might not be effective in patients with STAT3-HIES, although a few subsequent reports have reported successful outcomes. We update on progress of our patient now with over 18 years of follow-up and report on an additional seven cases, all of whom have survived despite demonstrating significant disease-related pathology prior to transplant. We conclude that effective cure of the immunological aspects of the disease and stabilization of even severe lung involvement may be achieved by allogeneic hematopoietic stem cell transplantation. Recurrent skin infections and abscesses may be abolished. Donor T_HIL17A to healthy controls. The future challenge will be to determine which patients should best be offered this treatment and at what point in their disease history.

Keywords: Autosomal dominant hyper IgE syndrome; Job syndrome; STAT3-HIES TH17 cells; dominant-negative STAT3 mutations; hematopoietic stem cell transplantation.

Background: Obesity is associated with a non-T2 asthma phenotype which is clinically more severe and difficult to control. Unfortunately, physicians have no gold standard biomarker to predict non-allergic asthma among obese patients.

Objective: To evaluate the ability of TNFA, IL17A and RORC mRNA expression to predict non-allergic asthma among obese adolescents.

Methods: This investigation was a cross-sectional study involving 4 adolescent groups: 1) healthy adolescents (n = 23), 2) adolescents with allergic asthma without obesity (n = 29), 3) adolescents with obesity without asthma (n = 28), and 4) adolescents with non-allergic asthma with obesity (n = 22).

Results: Receiver operating characteristic (ROC) curve analysis, the areas under the ROC curve (AUCs) for RORC, IL17A, and TNFA mRNA expression were 0.955 (95% CI: 0.86-0.99, p < 0.0001), 0.856 (95% CI: 0.73-0.94, p < 0.0001), and 0.756 (95% CI: 0.62-0.87, p = 0.0009), respectively.

Conclusion: RORC, IL17A and TNFA mRNA expression levels may be potential biomarkers of non-allergic asthma among obese adolescents in clinical practice.

Keywords: Asthma; IL-17A; Non-allergic asthma; Obesity; RORC; Tumour necrosis factor alpha.

This work is the first genetic association study of a potential relationship of single nucleotide polymorphisms rs8193036 and rs2275913 located in the IL17A promoter on chromosome 6p12 to chronic viral hepatitis and its progression in Kazakh population.

Evaluation of the effect of IL17A polymorphism on predisposition for chronic hepatitis B and C and its progression to liver cirrhosis.

A total of 862 individuals were enrolled in the retrospective case-control association study. Among the participants, 100 patients had chronic hepatitis B and/or C and liver cirrhosis, and 341 patients had chronic viral hepatitis only. Four hundred twenty-one (421) healthy HBV- and HCV-negative donors without liver diseases were recruited as population control. single nucleotide polymorphisms rs8193036[T/C] and rs2275913[G/A] were genotyped by TaqMan assays using genomic DNA extracted from peripheral blood cells. Results. Minor allele frequencies of rs8193036[C] and rs2275913[A] in the groups of patients were very similar to those observed in the control population, 0.4 and 0.3, respectively. Multivariate logistic regression analysis revealed odds ratios close to 1.0 and confidence intervals overlapping with the value of 1.0 and statistical significance p>> 0.4 for any groups under comparison in the multiplicative model of inheritance.

No significant association between two single nucleotide polymorphisms, rs8193036 and rs2275913 in the IL17A promoter, and susceptibility to chronic viral hepatitis C and/or B and disease progression to liver cirrhosis in Kazakh population were found.

Myelosuppression is the major dose-limiting toxicity of cancer chemotherapy. There have been many attempts to find new strategies that reduce myelosuppression. The dietary supplementation with lactic acid bacteria (LAB) improved respiratory innate immune response and the resistance against respiratory pathogens in immunosupressed hosts. Although LAB viability is an important factor in achieving optimal protective effects, non-viable LAB are capable of stimulating immunity. In this work, we studied the ability of oral preventive administration of viable and non-viable Lactobacillus rhamnosus CRL1505 or L. plantarum CRL1506 (Lr05, Lr05NV, Lp06V or Lp06NV, respectively) to minimize myelosuppressive and immunosuppressive effects derived from chemotherapy. Cyclophosphamide (Cy) impaired steady-state myelopoiesis in lactobacilli-treated and untreated control mice. Lr05V, Lr05NV and Lp06V treatments were the most effective to induce the early recovery of bone marrow (BM) tissue architecture, leukocytes, myeloid, pool mitotic and post-mitotic, peroxidase positive, and Gr-1^Low/High cells in BM. We selected the CRL1505 strain for being the one capable of maintaining its myelopoiesis-enhancing properties in its non-viable form. Although the CRL1505 treatments do not modify the Cy ability to induce apoptosis, both increased the incorporation of BrdU in BM cells. Consequently, Lr05NV and Lr05V treatments were able to promote early recovery of LSK cells (Lin^-Sca-1⁺c-Kit⁺ cells), multipotent progenitors (Lin^-Sca-1⁺c-Kit⁺CD34⁺ cells), and myeloid cells (Gr-1⁺Ly6G⁺Ly6C^- cells) with respect to the untreated Cy control. In addition, these treatments were able to increase the frequency of IL17A-producing innate lymphoid cells in the intestinal lamina propria (IL-17A⁺RORγt⁺CD4^-NKp46⁺ cells) after Cy injection. These results were correlated with an increase in the IL-17A serum levels, a GM-CSF high expression and a CXCL12 lower expression in BM. Therefore, both Lr05V and Lr05NV treatments are able to activate beneficially the IL-17A/GM-CSF axis and accelerate the recovery of Cy-induced immunosuppression by increasing BM myeloid precursors. We demonstrated for the first time the beneficial effect of CRL1505 strain on myelopoiesis affected by a chemotherapeutic drug. Furthermore, Lr05NV could be a good and safe resource for reducing chemotherapy-induced leukopenia. The results are a starting point for future research and open up broad prospects for future applications of the immunobiotics.

Keywords: cyclophosphamide; immunobiotic; lactobacilli; myelopoiesis; non-viable Lactobacillus.

T helper (Th) 17 cells protect from infections and are pathogenic in autoimmunity. While human Th17 cell differentiation has been defined, the global and stepwise transcriptional changes accompanying this process remain uncharacterized. Herein, by performing transcriptome analysis of human Th17 cells, we uncovered three time-regulated modules: early, involving exclusively "signaling pathways" genes; late, characterized by response to infections; and persistent, involving effector immune functions. To assign them an inflammatory or regulatory potential, we compared Th17 cells differentiated in presence or absence of interleukin (IL)-1β, respectively. Most inflammatory genes belong to the persistent module, whereas regulatory genes are lately or persistently induced. Among inflammatory genes, we identified the effector molecules IL17A, IL17F, IL26, IL6, interferon (IFN)G, IFNK, LTA, IL1A, platelet-derived growth factor (PDGF) A and the transcriptional regulators homeodomain-only protein homeobox (HOPX) and sex-determining-region-Y-box (SOX)2, whose expression was independently validated. This study provides an integrative representation of the stepwise human Th17 differentiation program and offers new perspectives toward therapeutic targeting of Th17-related autoimmune diseases.

Keywords: Immunology; Omics; Systems Biology.

Hyperforin is a major active constituent of Hypericum perforatum L. extract, which is widely used for the treatment of depressive disorders. Recent studies have reported that hyperforin reduced inflammation in stroke and suppressed proliferation and differentiation in keratinocytes. Psoriasis is a chronic immune-mediated inflammatory skin disease in which the IL-23/IL-17 axis plays an important role. To investigate the underlying inflammatory mechanisms and response of hyperforin in psoriasis, we use imiquimod (IMQ)-induced mice model, in vitro cultured murine splenic γδ T cells, and HaCaT cells in this study. Data showed that hyperforin reduced epidermal thickness and decreased IMQ-induced pathological scores of cutaneous skin lesions in mice. Meanwhile we proved that hyperforin suppressed infiltration of CD3⁺ T cells and downregulated expression of Il1, Il6, Il23, Il17a, Il22, antimicrobial peptides (AMPs) in the skin lesion. Hyperforin significantly inhibited imiquimod-induced splenomegaly, reduced serum levels of TNF-α and IL-6, and IL-17A in splenocytes and draining lymph nodes. Our study also suggested that hyperforin lessened the infiltration of γδ T cell and CCR6⁺ γδ T cells in spleen and lymph nodes. Hyperforin also suppressed the typical psoriasis-like inflammatory responses and the infiltration of IL-17A⁺ cells in dermal γδ T cells of IMQ treated Tcrd ^-/- mice transferred with γδ T cells. In vitro studies, hyperforin reduced the expression and secretion of IL-17A in γδ T cells, and suppressed the activation of MAPK/STAT3 pathways in human keratinocyte HaCaT cells and γδ T cells. In conclusion, hyperforin alleviates IMQ-induced inflammation in psoriasis through suppressing the immune responses exerted by IL-17 A-producing γδ T cells and related cytokines by modulating MAPK/STAT3 pathways. Our study provided a novel therapeutic tragedy for psoriasis by which hyperforin attenuates psoriasis-related inflammatory responses.

Keywords: IL-17A; Stat3; hyperforin; psoriasis; γδT cells.

The urgent need for a treatment of COVID-19 has left researchers with limited choice of either developing an effective vaccine or identifying approved/investigational drugs developed for other medical conditions for potential repurposing, thus bypassing long clinical trials. In this work, we compared the sequences of experimentally verified SARS-CoV-2 neutralizing antibodies and sequentially/structurally similar commercialized therapeutic monoclonal antibodies. We have identified three therapeutic antibodies, Tremelimumab, Ipilimumab and Afasevikumab. Interestingly, these antibodies target CTLA4 and IL17A, levels of which have been shown to be elevated during severe SARS-CoV-2 infection. The candidate antibodies were evaluated further for epitope restriction, interaction energy and interaction surface to gauge their repurposability to tackle SARS-CoV-2 infection. Our work provides candidate antibody scaffolds with dual activities of plausible viral neutralization and immunosuppression. Further, these candidate antibodies can also be explored in diagnostic test kits for SARS-CoV-2 infection. We opine that this in silico workflow to screen and analyze antibodies for repurposing would have widespread applications.

In the intestine, the host must be able to control the gut microbiota and efficiently absorb transiently supplied metabolites, at the risk of enormous infection. In mammals, the inflammatory cytokine interleukin (IL)-17A/F is one of the key mediators in the intestinal immune system. However, many functions of IL-17 in vertebrate intestines remain unclarified. In this study, we established a gene-knockout (KO) model of IL-17 receptor A1 (IL-17RA1, an IL-17A/F receptor) in Japanese medaka (Oryzias latipes) using genome editing technique, and the phenotypes were compared to wild type (WT) based on transcriptome analyses. Upon hatching, homozygous IL-17RA1-KO medaka mutants showed no significant morphological abnormality. However, after 4 months, significant weight decreases and reduced survival rates were observed in IL-17RA1-KO medaka. Comparison of gene-expression patterns in WT and IL-17RA1-KO medaka revealed that various metabolism- and immune-related genes were significantly down-regulated in IL-17RA1-KO medaka intestine, particularly genes related to mevalonate metabolism (mvda, acat2, hmgcs1, and hmgcra) and genes related to IL-17 signaling (such as il17c, il17a/f1, and rorc) were found to be decreased. Conversely, expression of genes related to cardiovascular system development, including fli1a, sox7, and notch1b in the anterior intestine, and that of genes related to oxidation-reduction processes including ugp2a, aoc1, and nos1 in posterior intestine was up-regulated in IL-17RA1-KO medaka. These findings show that IL-17RA regulated immune- and various metabolism-related genes in the intestine for maintaining the health of Japanese medaka.

The secretions of certain cytokines, chemokines and growth factors are triggered by orthodontic appliances, which often affect the remodelling of periodontal tissues. Critical cumulative forces are applied by various types of orthodontic appliances to the periodontium. The secretion of such molecules is probably responsible, through molecular and cellular communications, for the optimal resorption of hard tissues in the periodontal setting, which therefore enables the coordination of multiple movements of tooth. This study assessed and compared a wide range of cytokines, cellular marker analysis and defensins present in the saliva samples of human subjected to orthodontic treatment with two different treatment modalities, i.e., conventional lingual and labial fixed orthodontic appliances. A total 40 samples of saliva were obtained, of which 20 were treated with traditional lingual appliances and 20 were treated with labial fixed appliances. After 21 days of treatment, all salivary samples were collected from the subjects. In order to analyse a broad range of soluble cytokine levels in saliva by flow cytometry, a bead-based immunoassay was performed. Cell surface markers were analysed by flow cytometry. Protein levels of saliva for defensins were quantified by ELISA. Non-significant differences were observed in the cytokine levels in the saliva except for the significant effects for CCL2, IL-17A and IL-6. Cellular markers CD45 and CD326 showed high percentage in conventional lingual samples. Defensin levels were found to be lower in conventional lingual patients. Subjects with conventional lingual appliances had significantly higher salivary protein levels of IL-1β, CCL2, IL17A, and IL-6, higher CD45+ and CD326+ cells and lower defensin levels than subjects with fixed labial appliances. The current study provided a clear basis for the development of innovative methods to aid in the improvement of various procedural treatments and orthodontic equipment of next generation.

Psoriasis is a T cell mediated chronic inflammatory skin disease affecting about 2% of the population worldwide. Recently has established the central role of IL-23/Th17 immune axis in the pathogenesis of psoriasis and different subclasses of T cells including Th1 and Th17 cells are involved in initiation and amplification of the skin inflammation process, in addition, in cases of recurrent psoriasis, Th22 cells play the role of memory cells with the help of Th9 cells, which are also important in this process. The main goal was to evaluate the ratio of T cell profile and IL23/Th17 axis by evaluating IL17A, IL22, IL9 in peripheral blood of persons with moderate to severe plaque psoriasis. We have estimated the activation of IL-23/Th17 axis by evaluating the level of IL-17A, IL-22 and IL-9 in peripheral blood of patients with plaque psoriasis (n=18) with different severity of the disease (PASI from 10 to 40) comparing the results with data obtained from healthy persons (n=15). The expression of CD69 activation marker on T helper cells has been evaluated as well. The results were analyzed using FACScan flow cytometer (Becton Dickinson). The percentage of CD3 + T lymphocytes in the peripheral blood of patients with psoriasis was not significantly different compared to normal healthy volunteers, however, the level of expression of CD4 + T cells was reduced. We observed a dramatic increase in IL22 along with a decrease in the level of expression of IL-9 and IL-17, the expression of Th activation marker (CD69) was also decreased in comparison with the control group. The T cell profile and the IL-23/Th17 axis functional activity levels were significantly different from the literature data obtained about the inflammatory region (psoriatic lesions on the skin). IL-9 and IL-17 expression levels are decreased in peripheral blood Th cells, which may be explained by mobilization of the corresponding Th9 and Th17 cells into the inflammatory site.

Background: Interleukin (IL)17A is a member of the IL17 cytokine family, which is released by both immune and non-immune cells such as tumor and stromal cells into the tumor microenvironment. IL17 receptors are also widely expressed in different type of cells. Among all the members, IL17A is the most controversial in regulating tumor immunity. Here, we investigated how IL17A inhibition modulated macrophage differentiation and metabolism in the presence or absence of gemcitabine. Gemcitabine is the gold standard drug for treating pancreatic cancer and can increase macrophage antitumoral activities.

Results: We observed some unique features of macrophages polarized in the absence of IL17A, in terms of RNA and protein expression of typical phenotypic markers, and we demonstrated that this paralleled specific changes in their metabolism and functions, such as the induction of an antitumor response. Interestingly, these features were almost maintained or enhanced when macrophages were treated with gemcitabine. We also demonstrated that the anti-IL17A antibody effectively reproduced features of macrophages derived from IL17A knock-out mice.

Conclusion: Overall, we provide a proof-of-concept that combining an anti-IL17A antibody with gemcitabine may represent an effective strategy to modulate macrophages and enhance the anti-tumor response, especially in pancreatic cancer where gemcitabine is widely used.

Keywords: Interleukin 17A (IL17A); chemo-immunotherapy; macrophages; metabolism; pancreatic cancer.

HLA-B*27 is an important marker for spondyloarthritis (SpA), however, many SpA patients are HLA-B*27 negative. Thus, the aim of this study was to investigate the influence of IL17, TNF and VDR gene polymorphisms in SpA patients who were HLA-B*27 negative. This case-control study was conducted in 158 patients [102 patients with ankylosing spondylitis (AS) and 56 with psoriatic arthritis (PsA)] and 184 controls. HLA-B*27 genotyping was performed using PCR-SSP and IL17A (rs2275913), IL17F (rs763780), TNF-308 (rs1800629), TNF-238 (rs361525), FokI C>T (rs2228570), TaqI C>T (rs731236), ApaI A>C (rs7975232), and BsmI C>T (rs1544410) using PCR-RFLP. Statistical analyses were performed by Chi-square and logistic regression using OpenEpi and SNPStats software. The IL17F C allele frequency was higher in patients with SpA, AS and PsA compared to controls. The IL17F T/C genotype frequency was higher in SpA patients in an overdominant inheritance model and when men and women were separately analyzed. IL17A_IL17F AC haplotype was significantly associated to the risk for SpA patients. As for VDR, the ApaI a/a was a potential risk factor for SpA in men. In conclusion, IL17F C variant contributed to the risk of SpA in Brazilian patients who were HLA-B*27 negative and could be a potential marker for SpA.

Keywords: HLA-B27 antigen; IL17F C variant; ankylosing spondylitis; genetic association studies; psoriatic arthritis.

It is widely accepted that narcotic use during pregnancy and specific environmental factors (e.g., maternal immune activation and chronic stress) may increase risk of neuropsychiatric illness in offspring. However, little progress has been made in defining human-specific in utero neurodevelopmental pathology due to ethical and technical challenges associated with accessing human prenatal brain tissue. Here we utilized human induced pluripotent stem cells (hiPSCs) to generate reproducible organoids that recapitulate dorsal forebrain development including early corticogenesis. We systemically exposed organoid samples to chemically defined "enviromimetic" compounds to examine the developmental effects of various narcotic and neuropsychiatric-related risk factors within tissue of human origin. In tandem experiments conducted in parallel, we modeled exposure to opiates (μ-opioid agonist endomorphin), cannabinoids (WIN 55,212-2), alcohol (ethanol), smoking (nicotine), chronic stress (human cortisol), and maternal immune activation (human Interleukin-17a; IL17a). Human-derived dorsal forebrain organoids were consequently analyzed via an array of unbiased and high-throughput analytical approaches, including state-of-the-art TMT-16plex liquid chromatography/mass-spectrometry (LC/MS) proteomics, hybrid MS metabolomics, and flow cytometry panels to determine cell-cycle dynamics and rates of cell death. This pipeline subsequently revealed both common and unique proteome, reactome, and metabolome alterations as a consequence of enviromimetic modeling of narcotic use and neuropsychiatric-related risk factors in tissue of human origin. However, of our 6 treatment groups, human-derived organoids treated with the cannabinoid agonist WIN 55,212-2 exhibited the least convergence of all groups. Single-cell analysis revealed that WIN 55,212-2 increased DNA fragmentation, an indicator of apoptosis, in human-derived dorsal forebrain organoids. We subsequently confirmed induction of DNA damage and apoptosis by WIN 55,212-2 within 3D human-derived dorsal forebrain organoids. Lastly, in a BrdU pulse-chase neocortical neurogenesis paradigm, we identified that WIN 55,212-2 was the only enviromimetic treatment to disrupt newborn neuron numbers within human-derived dorsal forebrain organoids. Cumulatively this study serves as both a resource and foundation from which human 3D biologics can be used to resolve the non-genomic effects of neuropsychiatric risk factors under controlled laboratory conditions. While synthetic cannabinoids can differ from naturally occurring compounds in their effects, our data nonetheless suggests that exposure to WIN 55,212-2 elicits neurotoxicity within human-derived developing forebrain tissue. These human-derived data therefore support the long-standing belief that maternal use of cannabinoids may require caution so to avoid any potential neurodevelopmental effects upon developing offspring in utero.

Aims: Heart failure (HF) is common in both men and women, yet disease pathophysiology, presentation, and progression differ between sexes. Studies addressing whether biomarkers predict new onset HF sex-specifically are scarce. This study therefore aims to test the sex-specificity of 252 protein biomarkers for new-onset HF.

Methods and results: A matched case-control design in patients selected from cohorts within the HOMAGE consortium was used. Cases (new-onset HF, n = 562) and controls (n = 780) were matched for cohort (PREDICTOR, HEALTH-ABC, & PROSPER), follow-up time (defined as time from entry to incident HF), and age. Incident HF was defined as first hospitalization for HF. Targeted plasma proteins (n = 252) were measured using Proximity Extension Assay technology from O-link. To look for sex differences for new onset HF, we adjusted for cohort, age, and baseline clinical parameters. At baseline, women had a biomarker profile reflecting activated metabolism and immune responses. However, none of the biomarkers had a significant interaction with sex in predicting new onset HF, but four biomarkers had a trend towards sex-specificity (P < 0.013). E-selectin and interleukin 1 receptor antagonist were more female-specific, whereas IL17A and CHIT1 tended to be male sex-specific for incident HF.

Conclusions: The majority of biomarkers associated with incident HF did not significantly differ between women and men, despite clear differences in biomarkers at baseline.

Keywords: Incident heart failure; Proteomics; Sex differences.

Background/aim: The pathway of initiation of psoriasis comprises the differentiation and infiltration of T-helper 17 (Th17) cells into the skin, characterized by the production of interleukin 17A and 17F (IL-17A/IL-17F) among other cytokines, resulting in a downstream cascade of events. Due to the lack of simplicity in psoriasis models, we aimed to develop an easily and rapidly inducible mouse model for the IL-23/IL-17 pathway with quick readouts from a straightforward lavaging process and with detectable cytokine levels.

Materials and methods: We utilized the 6-day air-pouch mouse model, injected with a combination of anti-CD3, IL-23 and IL-1β. At 24, 48 and 72 h, intra-pouch secretion of IL-17A, IL-17F and C-X-C motif chemokine ligand 1 were measured. Skin biopsies were collected and immune cell infiltration evaluated, and intra-pouch immune cells were isolated and analyzed.

Results: The combination of anti-CD3, IL-23 with and without IL-1β significantly increased intra-pouch levels of IL-17A/IL-17F at 24 and 72 h, triggering a downstream production of C-X-C motif chemokine ligand 1. The cytokines were detectable even 72 h post-induction. T-cell receptor beta was down-regulated on CD4⁺ and CD8⁺ T-cells, indicating intra-pouch T-cell activation. Αnti-CD3 induced CD3⁺ cell migration into the subcutis and the lining tissue surrounding the cavity of the air pouch, where in the latter, a similar distribution pattern of Il17a mRNA-expressing cells was also observed. However, no Th17 cell differentiation nor changes in IL-17A⁺ granulocytes were observed.

Conclusion: The induced air-pouch mouse model induced with a cocktail of anti-CD3, IL-23 with or without IL-1β is able to mirror the IL-23/IL-17 axis of psoriasis-like inflammation characterized by immune cell infiltration and cytokine secretion.

Keywords: CXCL1; IL-17A; IL-17F; IL-1β; IL-23; Psoriasis; TCR; air pouch; anti-CD3.

Background: Canine leishmaniosis (CanL) is associated with an aberrant cutaneous immune response. Few studies have compared cutaneous immune cells among dogs with leishmaniosis or infected without leishmaniosis, and noninfected dogs.

Hypothesis/objective: Evaluate the number of neutrophils, histiocytes, T lymphocytes, T-bet-positive, GATA3-positive, FoxP3-positive and interleukin (IL)-17-positive cells, in lesional (Group A) and normal-looking (Group B) skin of nine dogs with stage II/III leishmaniosis; in normal-looking PCR-positive (Group C; n = 6) or PCR-negative (Group D; n = 6) skin of infected dogs; and in the normal-looking skin of 12 noninfected dogs (Group E).

Methods and materials: Diagnosis of CanL considered clinical signs, clinicopathological abnormalities, detection of Leishmania amastigotes in lymph nodes, and/or bone marrow and positive serological results. Paraffin-embedded skin biopsies were processed for routine immunofluorescence and positive cells were identified using commercially available anti-canine antibodies.

Results: In Group A, there was a significantly higher number of neutrophils (P < 0.001), histiocytes (P = 0.012), T lymphocytes (P = 0.011), GATA3-positive (P = 0.02) and IL-17A-positive (P = 0.002) cells compared to Group E. In Group B, there was a significantly higher number of histiocytes (P = 0.02), T lymphocytes (P = 0.004), GATA3-positive (P = 0.006) and FoxP3-positive (P = 0.028) cells compared to Group E. There was no difference in between groups A and B and between groups C or D and E.

Conclusions and clinical importance: In the lesional and/or normal-looking skin of dogs with moderate/severe CanL there is an infiltration of neutrophils, histiocytes, T lymphocytes, GATA3-, FoxP3- and IL17A-positive cells. By contrast, the number of these cells is not increased in the normal-looking skin of infected dogs without CanL compared to the skin of noninfected dogs.

Contexte: La leishmaniose canine (CanL) est associée avec une réponse immunitaire cutanée aberrante. Peu d’études ont comparé les cellules immunitaires cutanées entre les chiens avec leishmaniose ou infectés sans leishmaniose, et non infectés. HYPOTHÈSE/OBJECTIF: Evaluer le nombre de neutrophiles, histiocytes, lymphocytes T et cellules T-bet-positives, GATA3-positives, FoxP3-positives et interleukine (IL)-17 positives, dans la peau lésionnelle (Groupe A) et d’aspect normal (Groupe B) de neuf chiens avec leishmaniose stade II/III ; dans la peau d’aspect normal PCR-positive (Groupe C ; n=6) ou PCR-négative (Groupe D ; n=6) de chiens infectés ; et dans la peau d’aspect normal de 12 chiens non infectés (Groupe E). MATÉRIELS ET MÉTHODES: Le diagnostic de CanL repose sur les signes cliniques, les anomalies clinicopathologiques, la détection d’amastigotes de Leishmania dans les nœuds lymphatiques et/ou la moelle osseuse et les résultats sérologiques positifs. Les biopsies cutanées montées dans la paraffine ont été préparées pour immunofluorescence de routine et les cellules positives ont été mises en évidence par des anticorps anti-canin disponibles dans le commerce. RÉSULTATS: Dans le Groupe A, il y avait un nombre significativement plus élevé de neutrophiles (P < 0.001), histiocytes (P = 0.012), lymphocytes T(P = 0.011), cellules GATA3-positive (P = 0.02) et IL-17A-positive (P = 0.002) comparé au Groupe E. Dans le Groupe B, il y avait un nombre significativement plus élevé d’histiocytes (P = 0.02), lymphocytes T (P = 0.004), cellules GATA3-positive (P = 0.006) et FoxP3-positive (P = 0.028) comparé au Groupe E. Il n’y avait pas de différence entre les groupes A et B et entre les groupes C ou D et E.

Conlsusions et imprtance clinique: Au sein de la peau lésionnelle et/ou d’aspect normale de chiens avec CanL modére/sévère il y avait une infiltration de neutrophiles, histiocytes, lymphocytes T, cellules GATA-3, FOX-3 et IL17A positives. En revanche, le nombre de ces cellules n’est pas augmentée dans la peau d’aspect normal des chiens infectés sans CanL comparé à la peau de chiens non infectés.

INTRODUCCIÓN: la leishmaniosis canina (CanL) se asocia con una respuesta inmunitaria cutánea aberrante. Pocos estudios han comparado las células inmunitarias cutáneas entre perros con leishmaniosis o infectados sin leishmaniosis y perros no infectados. HIPÓTESIS/OBJETIVO: Evaluar el número de neutrófilos, histiocitos, linfocitos T, células T-bet-positivas, GATA3-positivas, FoxP3-positivas e interleuquina (IL)-17-positivas, enzonas de piel lesionadas (Grupo A) y de apariencia normal (Grupo B) piel de nueve perros con leishmaniosis en estadio II/III; en la piel de perros infectados con PCR positiva (Grupo C; n = 6) o PCR negativa (Grupo D; n = 6) de aspecto normal; y en la piel de aspecto normal de 12 perros no infectados (Grupo E). MÉTODOS Y MATERIALES: el diagnóstico de CanL consideró signos clínicos, anomalías clínico-patológicas, detección de amastigotes de Leishmania en ganglios linfáticos y/o médula ósea y resultados serológicos positivos. Las biopsias de piel incluidas en parafina se procesaron para inmunofluorescencia de rutina y las células positivas se identificaron utilizando anticuerpos anti-caninos disponibles comercialmente. RESULTADOS: en el grupo A, hubo un número significativamente mayor de neutrófilos (P <0,001), histiocitos (P = 0,012), linfocitos T (P = 0,011), célula positivas para GATA3 (P = 0,02) e IL-17A positivas (P = 0,002) en comparación con el Grupo E. En el Grupo B, hubo un número significativamente mayor de histiocitos (P = 0,02), linfocitos T (P = 0,004), células GATA3 positivas (P = 0,006) y FoxP3 positivas (P = 0,028) en comparación con el Grupo E. No hubo diferencias entre los grupos A y B ni entre los grupos C o D y E. CONCLUSIONES E IMPORTANCIA CLÍNICA: en la piel lesionada y/o de aspecto normal de perros con CanL moderada/grave hay una infiltración de neutrófilos, histiocitos, linfocitos T, células positivas para GATA3, FoxP3 e IL17A. Por el contrario, el número de estas células no aumenta en la piel de aspecto normal de los perros infectados sin CanL en comparación con la piel de los perros no infectados.

Hintergrund: Die canine Leishmaniose (CanL) wird mit einer abnormen kutanen Immunantwort in Verbindung gebracht. Wenige Studien haben die kutanen Immunzellen bei Hunden mit Leishmaniose oder infizierten ohne Leishmaniose und nicht-infizierten Hunden verglichen.

Hypothese/ziele: Eine Evaluierung der Neutrophilen, Histiozyten, T Lymphozyten, T-bet-positiven, GATA3-positiven, FoxP3-positiven und Interleukin (IL)-17-positiven Zellen, in Hautveränderungen (Gruppe A) und normal aussehender Haut (Gruppe B) von neun Hunden mit Stadium II/III der Leishmaniose; in einer Gruppe mit normal aussehender PCR-positiver (Gruppe C; n = 6) oder PCR-negativer (Gruppe D; n = 6) Haut von infizierten Hunden; sowie in normal aussehender Haut von 12 nicht-infizierten Hunden (Gruppe E).

Methoden und materialien: Für eine Diagnose von CanL wurden klinische Zeichen, klinisch-pathologische Abnormalitäten, die Detektion von Leishmania Amastigoten im Lymphknoten, und/oder Knochenmark sowie positive serologische Ergebnisse in Betracht gezogen. In Paraffin-gebettete Hautbiopsien wurden für Routine Immunfluoreszenz weiterverarbeitet und positive Zellen mittels kommerziell erhältlichen anti-caninen Antikörpern identifiziert.

Ergebnisse: In Gruppe A gab es eine signifikant höhere Anzahl an Neutrophilen (P < 0,001), Histiozyten (P = 0,012), T Lymphozyten (P = 0,011), GATA3-positiven (P = 0,02) und IL-17A-positiven (P = 0,002) Zellen im Vergleich zu Gruppe E. In Gruppe B bestand eine signifikant höhere Anzahl an Histiozyten (P = 0,02), T Lymphozyten (P = 0,004), GATA3-positiver (P = 0,006) und FoxP3-positiver (P = 0,028) Zellen im Vergleich zu Gruppe E. Es bestand kein Unterschied zwischen Gruppen A und B und zwischen den Gruppen C oder D und E.

Schlussfolgerungen und klinische bedeutung: In der läsionalen und/oder normal-aussehenden Haut von Hunden mit moderaten/schweren CanL kommt eine Infiltration mit Neutrophilen, Histiozyten, T Lymphozyten, GATA3-, FoxP3- und IL17A-positiven Zellen vor. Im Vergleich dazu war die Anzahl dieser Zellen in der normal aussehenden Haut der infizierten Hunde ohne CanL im Vergleich zur Haut nicht-infizierter Hunde nicht erhöht.

背景: 犬リーシュマニア症 (CanL) は、異常な皮膚免疫反応に関連している。リーシュマニア症の犬またはリーシュマニア症なしで感染した犬と感染していない犬の間で皮膚免疫細胞を比較した研究はほとんどない。 仮説/目的: 本研究の目的は、ステージII / IIIのリーシュマニア症の9頭の犬の病変皮膚 (グループA) および正常に見える皮膚 (グループB) 、感染犬の正常に見えるPCR陽性の皮膚 (グループC; n = 6) またはPCR陰性の皮膚 (グループD; n = 6) 、そして感染していない12頭の犬の正常に見える皮膚 (グループE) の好中球、組織球、Tリンパ球、T-bet陽性、GATA3陽性、FoxP3陽性、インターロイキン (IL) -17陽性細胞の数を評価することであった。 材料と方法: CanLの診断では、臨床徴候、臨床病理学的異常、リンパ節におけるリーシュマニアアマスティゴートの検出、および/または骨髄および血清学的結果陽性が考慮された。パラフィン包埋皮膚生検をルーチン免疫蛍光法のため処理し、市販抗イヌ抗体を使用して陽性細胞を同定した。 結果: グループAはグループEと比較して、好中球 (P <0.001) 、組織球 (P = 0.012) 、Tリンパ球 (P = 0.011) 、GATA3陽性 (P = 0.02) 、IL-17A陽性 (P = 0.002) の数が有意に多かった。グループBはグループEと比較して、組織球 (P = 0.02) 、Tリンパ球 (P = 0.004) 、GATA3陽性 (P = 0.006) 、FoxP3陽性 (P = 0.028の数が有意に多かった。グループAB間、およびグループCまたはDE間では相違なかった。 結論と臨床的重要性: 中等度/重度のCanL犬の病変および/または正常に見える皮膚には、好中球、組織球、Tリンパ球、GATA3、FoxP3、およびIL17A陽性細胞の浸潤がある。対照的に、これらの細胞数は、感染していない犬の皮膚と比較して、CanLのない感染犬の正常に見える皮膚では増加しない。.

背景: 犬利什曼病(CanL)可引起皮肤免疫应答异常。很少有研究比较利什曼病患犬或无利什曼病的感染犬和非感染犬的皮肤免疫细胞。 假设/目的: 评价9只II/III期利什曼病犬, 其病变 (A组) 和外观正常 (B组) 的皮肤的中性粒细胞、组织细胞、T淋巴细胞、T-bet-阳性、GATA3阳性、FoxP3阳性和白介素(IL)-17阳性细胞数量; 外观正常的PCR阳性 (C组; n=6) 或PCR阴性 (D组; n=6) 的感染犬皮肤; 以及12只非感染犬 (E组) 外观正常的皮肤。 方法和材料: CanL的诊断考虑了临床体征、临床病理学异常、淋巴结和/或骨髓中利什曼原虫无鞭毛体的检测和阳性血清学结果。对石蜡包埋的皮肤活检组织进行常规免疫荧光处理, 并使用市售抗犬抗体鉴定阳性细胞。 结果: A组的中性粒细胞(P<0.001)、组织细胞(P=0.012)、T淋巴细胞(P=0.011)、GATA3阳性(P=0.02)和IL-17A阳性(P=0.002)细胞数量显著高于E组。B组的组织细胞数量显著更高(P = 0.02), 与E组相比, T淋巴细胞(P=0.004)、GATA3阳性(P=0.006)和FoxP3阳性(P=0.028)细胞。A组和B组之间,以及C组或D组和E组之间无差异。 结论和临床重要性: 在中度/重度CanL犬的病变和/或外观正常的皮肤中, 存在中性粒细胞、组织细胞、T淋巴细胞、GATA3、FoxP3和IL17A阳性细胞浸润。相比之下, 与未感染犬的皮肤相比, 在无CanL的感染犬的正常外观皮肤中,这些细胞的数量未增加。.

Contexto: A leishmaniose canina (CanL) está associada a uma resposta imune cutânea aberrante. Poucos estudos compararam células imunes cutâneas entre cães com leishmaniose ou infectados sem leishmaniose e cães não infectados. HIPÓTESE/OBJETIVO: Avaliar o número de neutrófilos, histiócitos, linfócitos T, células T-bet-positivas, GATA3-positivas, FoxP3-positivas e interleucina (IL) -17-positivas, em pele lesional (Grupo A) e de aparência normal (Grupo B) de nove cães com leishmaniose em estágio II / III; em pele de cães infectados positiva por PCR de aparência normal (Grupo C; n = 6) ou negativa por PCR (Grupo D; n = 6); e na pele de aparência normal de 12 cães não infectados (Grupo E). MÉTODOS E MATERIAIS: O diagnóstico do CanL foi baseado nos sinais clínicos, anormalidades clínico-patológicas, detecção de formas amastigotas de Leishmania em linfonodos e/ou medula óssea e resultados sorológicos positivos. As biópsias de pele incluídas em blocos de parafina foram processadas para imunofluorescência de rotina e as células positivas foram identificadas usando anticorpos anticaninos disponíveis no mercado.

Resultados: No Grupo A, houve um número significativamente maior de neutrófilos (P <0,001), histiócitos (P = 0,012), linfócitos T (P = 0,011), células GATA3-positivas (P = 0,02) e IL-17A-positivas ( P = 0,002) em comparação com o Grupo E. No Grupo B, houve um número significativamente maior de histiócitos (P = 0,02), linfócitos T (P = 0,004), células GATA3-positivas (P = 0,006) e FoxP3-positivas (P = 0,028) em comparação com o Grupo E. Não houve diferença entre os grupos A e B e entre os grupos C ou D e E. CONCLUSÕES E IMPORTÂNCIA CLÍNICA: Na pele com lesões e/ou aparência normal de cães com CanL moderada/grave há infiltração de neutrófilos, histiócitos, linfócitos T, células GATA3-, FoxP3- e IL17A-positivas. Em contraste, o número dessas células não está aumentado na pele de aparência normal de cães infectados sem CanL em comparação com a pele de cães não infectados.

Background and aims: Expansion and morphological dysregulation of the bronchial vascular network occurs in asthmatic airways. Interleukin (IL) -17 and Rho-kinase (ROCK) are known to act in inflammation control and remodeling. Modulation of Rho-kinase proteins and IL-17 may be a promising approach for the treatment of asthma through the control of angiogenesis. Our objective was to analyze the effects of treatment with anti-IL17 and/or Rho-kinase inhibitor on vascular changes in mice with chronic allergic pulmonary inflammation.

Methods: Sixty-four BALB/c mice, with pulmonary inflammation induced by ovalbumin were treated with anti-IL17A (7.5/µg per dose, intraperitoneal) and/or Rho-kinase inhibitor (Y-27632-10 mg/kg, intranasal), 1 h before each ovalbumin challenge (22, 24, 26, and 28/days). Control animals were made to inhale saline. At the end of the protocol, lungs were removed, and morphometric analysis was performed to quantify vascular inflammatory, remodeling, and oxidative stress responses.

Results: Anti-IL17 or Rho-kinase inhibitor reduced the number of CD4⁺, CD8⁺, dendritic cells, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17, Rho-kinase 1 and 2, transforming growth factor (TGF-β), vascular endothelial growth factor (VEGF), nuclear factor (NF)-KappaB, iNOS, metalloproteinase (MMP)-9, MMP-12, metalloproteinase inhibitor-1 (TIMP-1), FOXP-3, signal transducer and activator of transcription 1 (STAT1) and phospho-STAT1-positive cells, and actin, endothelin-1, isoprostane, biglycan, decorin, fibronectin and the collagen fibers volume fraction compared with the ovalbumin group (p < 0.05). The combination treatment, when compared with anti-IL17, resulted in potentiation of decrease in the number of IL1β- and dendritic cells-positive cells. When we compared the OVA-RHO inhibitor-anti-IL17 with OVA-RHO inhibitor we found a reduction in the number of CD8⁺ and IL-17, TGF-β, and phospho-STAT1-positive cells and endothelin-1 in the vessels (p < 0.05). There was an attenuation in the number of ROCK 2-positive cells in the group with the combined treatment when compared with anti-IL17 or Rho-kinase inhibitor-treated groups (p < 0.05).

Conclusion: We observed no difference in angiogenesis after treatment with Rho-kinase inhibitor and anti-IL17. Although the treatments did not show differences in angiogenesis, they showed differences in the markers involved in the angiogenesis process contributing to inflammation control and vascular remodeling.The reviews of this paper are available via the supplemental material section.

Keywords: Rho-kinase; Y-27632; angiogenesis; asthma; interleukin-17; neutralizing antibody; vascular remodeling.

Purpose: To investigate the immuno-histological evidences in viable and non-viable hydatid cysts obtained from naturally infected camels.

Methods: A cohort study (February 2018-December 2019), a total of 15 hydatidosis-infected camels from slaughter houses in Cairo were involved. Specimens were investigated for parasite viability, liver histological changes, IL-17A cytokine immunohistochemical expressions in the adventitial layer, and the anti-nuclear antibodies (ANAs) immunofluorescent expression in the metacestode's structures. Real-Time Quantitative -Morphocytometry and SPSS were utilized.

Results: Multi-focal lesions and high viability were found in 60% of the cases. Overall accumulation of collagen associated the parasite establishment that involved infiltrations of mononuclear cells with significantly increased IL-17A expression. Interestingly, the ANAs appeared to have a role in the immune-defense against the metacestode showing different patterns. ANAs production correlated with IL-17A expression and the viability of the parasite.

Conclusion: IL-17A responses in hydatidosis is associated with collagen deposition and ANA production as a sort of anti-parasite immunity in a viability dependent manner.

Keywords: ANA; Collagen; IL17A; Viability.