Intraepithelial T-cell Research Articles

Analysis of off-tumour toxicities of T-cell-engaging bispecific antibodies via donor-matched intestinal organoids and tumouroids

Predicting the toxicity of cancer immunotherapies preclinically is challenging because models of tumours and healthy organs do not typically fully recapitulate the expression of relevant human antigens. Here we show that patient-derived intestinal organoids and tumouroids supplemented with immune cells can be used to study the on-target off-tumour toxicities of T-cell-engaging bispecific antibodies (TCBs), and to capture clinical toxicities not predicted by conventional tissue-based models as well as inter-patient variabilities in TCB responses. We analysed the mechanisms of T-cell-mediated damage of neoplastic and donor-matched healthy epithelia at a single-cell resolution using multiplexed immunofluorescence. We found that TCBs that target the epithelial cell-adhesion molecule led to apoptosis in healthy organoids in accordance with clinical observations, and that apoptosis is associated with T-cell activation, cytokine release and intra-epithelial T-cell infiltration. Conversely, tumour organoids were more resistant to damage, probably owing to a reduced efficiency of T-cell infiltration within the epithelium. Patient-derived intestinal organoids can aid the study of immune–epithelial interactions as well as the preclinical and clinical development of cancer immunotherapies.

Low dietary fiber intake impairs small intestinal Th17 and intraepithelial T cell development over generations

Dietary fiber strongly impacts the microbiota. Here, we show that a low-fiber diet changes the small intestinal (SI) microbiota and impairs SI Th17, TCRαβ+CD8αβ+ and TCRαβ+CD8αα+ intraepithelial Tcell development. We restore Tcell development with dietary fiber supplementation, but this defect becomes persistent over generations with constant low-fiber diets. Offspring of low-fiber diet-fed mice have reduced SI Tcells even after receiving a fiber-rich diet due to loss of bacteria important for Tcell development. In these mice, only a microbiota transplant from a fiber-rich diet-fed mouse and a fiber-rich diet can restore Tcell development. Low-fiber diets reduce segmented filamentous bacteria (SFB) abundance, impairing its vertical transmission. SFB colonization and a fiber-rich diet partially restore Tcell development. Finally, we observe that low-fiber diet-induced Tcell defects render mice more susceptible to Citrobacter rodentium infection. Together, these results demonstrate the importance of fiber to microbiota vertical transmission and host immune system development.

Specific T-cell receptor beta-rearrangements of gluten-triggered CD8+ T-cells are enriched in celiac disease patients' duodenal mucosa

Celiac disease (CeD) is an autoimmune disorder affecting the small intestine with gluten as disease trigger. Infections including Influenza A, increase the CeD risk. While gluten-specific CD4+ T-cells, recognizing HLA-DQ2/DQ8 presented gluten-peptides, initiate and sustain the celiac immune response, CD8+ α/β intraepithelial T-cells elicit mucosal damage. Here, we subjected TCRs from a cohort of 56 CeD patients and 22 controls to an analysis employing 749 published CeD-related TCRβ-rearrangements derived from gluten-specific CD4+ T-cells and gluten-triggered peripheral blood CD8+ T-cells. We show, that in addition to TCRs from gluten-specific CD4+ T-cells, TCRs of gluten-triggered CD8+ T-cells are significantly enriched in CeD duodenal tissue samples. TCRβ-rearrangements of gluten-triggered CD8+ T-cells were even more expanded in patients than TCRs from gluten-specific CD4+ T-cells (p < 0.0002) and highest in refractory CeD. Sequence alignments with TCR-antigen databases suggest that a subgroup of these most likely indirectly gluten-triggered TCRs recognize microbial, viral, and autoantigens.

Update on the Pathogenesis of Enteropathy-Associated T-Cell Lymphoma.

EATL is an aggressive T-cell non-Hodgkin lymphoma with poor prognosis and is largely localized to the small intestine. EATL is closely associated with coeliac disease (CD) and is seen mostly in patients originating from Northern Europe. Various factors are associated with an increased risk of developing EATL, such as viral infection, advanced age, being male, and the presence of the HLA-DQ2 haplotype. Clonal rearrangements in the TCR-β and γ genes have been reported in all EATL morphological variants with distinctive immunophenotypic characteristics. Although EATL can occur de novo, individuals with RCDII are at a higher risk of developing EATL. The cells of origin of EATL has been postulated to be normal small intestinal intraepithelial T-lymphocytes (IELs), and more recent evidence suggests a link between innate precursor IELs and EATL derived from refractory coeliac disease type II (RCDII). The immune microenvironment of mucosal cells within the small intestine enhances the process of neoplastic transformation of IELs into EATL. Cytokines such as IL-15 can activate and crucially deregulate the JAK-STAT signaling pathway by binding to receptors on the surface of IELs. Furthermore, mutations in the JAK/STAT pathway have been associated with RCDII-derived EATL.

Immunopathology of early and advanced epididymis lesions caused by Brucella ovis in rams

Ovine brucellosis is an infectious disease that causes alterations in the reproductive tract in ram and abortion in ewes. Their negative economic impact in ovine production warrants a thorough understanding the interactions between B. ovis and the host. Here, epididymis lesions of rams infected by B. ovis were histopathologically staged into early and advanced. Expression by immunohistochemistry of Brucella antigens, inflammatory cell markers (CD3, CD79αcy) and cytokines (IFN-γ, TNF-α, TGF-β1) was assessed in both stages. Early lesions were characterized by epithelial changes, interstitial inflammation, and mild fibrosis; whereas advanced lesions displayed caseous granulomas containing numerous macrophages, multinucleated giant cells, lymphocytes, and plasma cells. Expression of Brucella antigens were observed in both stages. The cellular response in B. ovis lesions were predominantly of T-cells (CD3+) whereas low numbers of B-cells and plasma cells (CD79αcy+) were present in both early and advanced lesions. IFN-γ was expressed by lymphocytes in early lesions suggesting that the adaptive immune response against B. ovis is initiated by Th1 cells, this response was also preserved in advanced stages. Expression of TNF-α was observed in neutrophils of epithelial microabscesses and intraepithelial T-cells of early lesions suggesting a promotion of neutrophil phagocytosis triggered by TNF-α. On the other hand, advanced lesions showed a reduction of TNF-α expression which may permit B. ovis persistence in granulomas. Lastly, TGF-β1 expression (fibroblast, macrophages and less in lymphocytes) were increased with time, suggesting that B. ovis promotes TGF-β1 secretion promoting chronicity of the lesions.

Interferon regulatory factor-2 is required for the establishment of the gut intraepithelial T-cell compartment.

CD8αα+ intestinal intraepithelial lymphocytes (iIELs) are known for their unique role in keeping the integrity of the intestinal epithelial barrier, but factors affecting the development of these cells have not been thoroughly understood. Here, we found that the transcriptional regulator interferon regulatory factor-2 (IRF-2) plays a cell-intrinsic, indispensable role in establishing iIEL populations. CD8αα+, but not CD8αβ+, iIELs bearing TCRαβ or TCRγδ were severely reduced in numbers in mice lacking this factor (Irf2-/- mice). Moreover, the majority of residual CD8αα+TCRαβ+ iIELs in these mice was immature as judged from their Thy1.2high phenotype and inefficient T-bet expression. Thymic IEL precursors isolated from Irf2-/- mice failed to efficiently generate CD8αα+TCRαβ+ and TCRγδ+ IELs upon transfer in vivo and CD8αα+TCRαβ+ cells in response to IL-15 in vitro. Double mutant mice lacking both interleukin-15 (IL-15) and IRF-2 showed an even more severe iIEL defect than in mice lacking IL-15 alone. Upon increasing agonistic TCR signal strength through OT-II TCR transgenesis, CD8αα+TCRαβ+ iIELs became more abundant but remained immature on the Irf2-/- background. Our current observations, thus, revealed the unique bimodal role that IRF-2 plays in promoting not only generation of IEL progenitors in the thymus but also maturation of iIELs in the periphery in IL-15-dependent and -independent manners.

Microscopic anatomical background of unexplained tubal infertility – three new possible causes

Infertility is defined as the inability to conceive within 1 year of unprotected intercourse. The infertility evaluation generally includes an assessment of the ovarian cycle and a semen analysis, but the evaluation of tubal patency is an essential part of the fertility workup. However, in the case of the uterine tube, patency may not be the only cause of tubal infertility. Idiopathic or unexplained infertility is a broad subcategory of infertility defined by the absence of obvious or explainable cause. Based on our previous research, we review and discuss three hypotheses associated with new anatomical and histological knowledge of tubal morphology and transport function and its potential impact on reproductive outcomes in women with unexplained tubal infertility. In 2021, Duffy et al. (Fertility and Sterility) published “Top 10 priorities for future infertility research”, based on their survey among healthcare professionals and patients with fertility problems from 40 countries. No mention of tubal infertility was included in their results. On the other hand, the topic “What causes unexplained infertility?” was one of the most important research priorities for female infertility. In 2018, a neglected structure of the human uterine tube, the lymphatic lacunae of the mucosal folds and fimbriae were described by Varga et al. (Annals of Anatomy). We believe that the lymph and lymphatic lacunae are responsible for the thickening of the fimbriae during ovulation. The fimbriae become enlarged and turgescent, coming into contact with the surface of the ovary, what is important in oocyte pick‐up and the maintenance of the tubal fluid. Hypothetically, disturbed lymph flow, caused by e.g., sedentary work, can cause problems with oocyte pick‐up and subsequent fertilization. In 2019, Varga et al. (Annals of Anatomy) described an important regulatory T‐ cell population in the human tubal epithelium. These intra‐epithelial T‐lymphocytes can be involved in the process of immune tolerance, which could lead to the tolerance of non‐self cells (sperm) and partially non‐self cells (a developing embryo) without the activation of local immune responses. An imbalance between the cells of the intra‐epithelial immune system can hypothetically lead to the immune reaction against sperm and/or embryo during transport through the uterine tube. The recently described cell population of tubal telocytes have many possible functions, but one of the most commonly reported is their ability to “sense” the levels of female sex hormones and thus regulate those processes inside the uterine tubes, which are under hormonal control. As reviewed by Klein et al. (2020, Medicina), telocytes are excessively fragile in the presence of noxious factors. In the uterine tubes, possible mechanisms of unexplained infertility include changes in the structural and functional integrity of the cellular microenvironment, including quantitative reduction and/or functional alteration of telocytes.

Abstract 154: Endothelial Dysfunction In Experimental Celiac Disease Is Mediated By Gut-Derived Vascular Inflammation And Oxidative Stress

Objective: Celiac disease (CeD) is an autoimmune enteropathy triggered by dietary gluten, with a prevalence around 1%. Since many patients show heterogeneous symptoms, it is vastly underdiagnosed and yet without any treatment option besides lifelong gluten restriction. Vascular function in CeD patients compared to healthy controls has been assessed and hypothesized CeD as a cardiovascular risk factor. Herewith we aimed to explore the cardiovascular effects of CeD in a murine model, with special emphasis on endothelial dysfunction, vascular inflammation, and oxidative stress, known triggers for cardiovascular disease. Methods: NOD.DQ8 mice were raised on a zein-based, gluten-free diet. During the experiment, mice received gavage with peptic-tryptic digest of gliadin and were switched to a gluten-containing diet (Gliadin group). A control group (Zein group) was continued on gluten-free diet and received accordingly digested zein. Results: Blood pressure measurements revealed arterial hypertension in the Gliadin group. Endothelium-dependent relaxation was impaired in the Gliadin group compared to Zein, whereas no change in endothelium-independent was observed, tested by isometric tension recordings. Pro-inflammatory genes, like Tnfa and Nox2 were upregulated in the Gliadin group measured by quantitative rtPCR in aortic and intestinal tissue. Infiltration by intraepithelial T-lymphocytes into the epithelium of the small intestine was shown by IHC. In cardiac tissue, gliadin-treatment increased the oxidative stress parameters 3-nitrotyrosine and 4-hydroxy-2-nonenal shown in dot blots. To investigate the inflammatory link between gut and the cardiovascular system plasma proteomics were performed and revealed elevated levels of IL-17A. Conclusion: CeD is a cardiovascular risk factor. However, the mechanism remains unresolved. In a mouse model, we demonstrate that gluten treatment elevates blood pressure and impairs vascular function. Intestinal inflammation conveys to the cardiovascular system, leads to oxidative stress, and thereby impairs endothelial function. A potential mediator might be IL-17A, whose role in other autoimmunity-based cardiovascular risk factors (e.g., psoriasis) has already been demonstrated.

Microanalysis of the Intestinal Bulb of Grass Carp (Ctenopharyngodon Idella): Histological, Histochemical, Immunohistochemical, and Scanning Electron Microscopical Studies.

Cyprinid fishes have one of the simplest types of gastrointestinal tract among vertebrates. Those fish species do not possess a true stomach that is replaced by a simple dilatation at the anterior part of the intestine called the intestinal bulb. Twenty adult specimens of grass carp were used in the present study to identify the cellular components as well as the immunohistochemical and surface architectural characteristics of the intestinal bulb. The mucosa of the intestinal bulb shows numerous, deep longitudinal folds arranged in zigzagging-like patterns. The epithelium is composed mainly of absorptive columnar cells covered by microvilli and mucous goblet cells. Spindle-shaped enteroendocrine cells and some migratory immune cells such as intraepithelial lymphocytes and rodlet cells could be identified between the absorptive cells. The epithelium also contains many secretory granules and large numbers of vacuoles containing digestive enzymes mostly in the basal part. The immunohistochemistry revealed that CD20-positive B-lymphocytes are immunolocalized mainly in the connective tissue core lamina propria of the mucosal folds. However, CD3-immunopositive T-lymphocytes are highly concentrated in the lamina propria. In addition, intraepithelial T-lymphocytes expressed immunopositivity to CD3. The current study presented many types of immune cells and suggests their essential immunological role for the intestinal blub.

Challenge in Early Diagnosis of Rare Lymphomas – A Case of Monomorphic Epitheliotropic Intestinal T-cell Lymphoma Masquerading as Enteritis

Abstract Introduction/Objective Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL) is an extranodal lymphoma arising from intestinal intraepithelial T-lymphocytes. Diagnosis is challenging obscured by non-specific gastrointestinal (GI) symptoms and radiographic findings, with potential delay in implementing therapy, as was encountered in a 40- year-old man of African-American descent with cachexia. Methods/Case Report The patient presented with a two month history of nausea, vomiting, diarrhea and 10-15 pounds weight loss. Computed tomography revealed wall thickening of multiple large- and small-bowel loops. The clinical differential diagnoses included infectious enteritis and inflammatory bowel disease. Since symptoms improved during the hospital stay, he was discharged with future outpatient endoscopy planned but returned a month later with abdominal swelling and pain. On repeat imaging, he had ascites and small-bowel mechanical obstruction versus ileus. Straw-colored ascites, dilated friable segments of small-bowel, matted mesenteric lymph nodes, and adhesions were found on exploratory laparotomy. Small-bowel segments and appendix were resected. A prolonged hospital course ensued post-operatively with death due to multiple complications. Results (if a Case Study enter NA) The intestinal wall was grossly thickened with no discrete masses and microscopically had extensive infiltrates of small- to intermediate-sized, mature, minimally pleomorphic, CD3+/CD4- /CD5-/CD7+/CD8+/CD10-/CD20-/CD30-/CD56+/EBER-ISH- T-lymphocytes, focally infiltrating into the epithelium. Ascitic fluid cytology had small lymphocytes, CD2+/CD3+/CD4-/CD5-/CD10-/CD7+/CD8+/CD25-/CD30-/CD56+ /TCRαβ+/TCRγδ- by flow cytometry immunophenotyping. A diagnosis of MEITL was rendered based on these findings. Conclusion MEITL is rare, aggressive, unassociated with celiac disease and usually seen in Asian and Hispanic populations. It presents with non-specific GI symptoms of abdominal pain, perforation, diarrhea, weight loss and intestinal obstruction underscoring the need for a high index of suspicion. Upper GI endoscopy has a limited role since involvement of small-bowel, specifically jejunum, is common. The evolving capsule endoscopy techniques may help in an early diagnosis. Due to limited literature, optimal treatment is unclear and various strategies using chemotherapy, surgery and stem-cell transplant have been tried with variable outcomes.

Management of mixed acute rejection driven by a de novo donor-specific complement-binding anti-DQB1*03:01 antibody and intraepithelial CD8 T cells in a kidney recipient: A case report

Mixed acute rejection is a clinicopathological entity that is difficult to accurately diagnose, and so may be under-reported. Allografts are lost more often than in either humoral or cellular rejection. The diagnosis requires both histological and immunological studies on renal biopsy and blood specimens from the transplant recipient to provide the required rescue therapy to abolish the allogeneic response against the graft. We present a clinical case report of an active mixed acute rejection driven by a de novo donor-specific complement-binding anti-DQB1*03:01 antibody and intraepithelial CD8 T-cells in a patient with a kidney transplant. The patient was diagnosed, treated, and followed up as per the local institution’s procedure with a full recovery of graft function. Our case emphasises the challenge of a mixed acute rejection and supports the view of the need to improve the post-transplant outcome of recipients and their grafts.

Diagnostic and therapeutic challenge of unclassifiable enteropathies with increased intraepithelial CD103+ CD8+ T lymphocytes: a single center case series

Objectives Chronic diarrhea, villous atrophy and/or increased intraepithelial T-lymphocytes (IEL) occur in many inflammatory disorders including celiac disease (CD). However, a definite diagnosis is difficult to make in some patients despite an extensive diagnostic work-up. Clinical outcomes and histological phenotypes of such patients we refer to as unclassifiable enteropathy (UEP) remain unclear. Material and methods We performed a retrospective single-center analysis of patients with chronic diarrhea, weight loss and increased IEL. Patients with defined etiologies including infections, CD, drugs, immunodeficiencies or neoplasms were excluded. Clinical and histologic/immunophenotypic parameters were analyzed. Results Nine patients with UEP were identified. Small intestinal damage ranged from minor villous abnormalities to complete atrophy while all patients displayed high numbers of CD103+ CD8+ IELs. In contrast to CD, these CD8+ T cells were not confined to the surface epithelium, but also infiltrated the crypts. Additional histological features included apoptotic crypt epithelial cells and mixed inflammatory infiltrates in the tunica propria. Involvement of other segments of the gastrointestinal tract was observed in 7/9 patients. A clonal intestinal T-cell lymphoproliferative disorder developed in 2 patients, one of which had a fatal disease course. The majority of patients responded to corticosteroids, while response to immunosuppressive medications yielded heterogeneous results. Conclusions We report a patient population with ‘difficult-to-classify’ enteropathies characterized by various degrees of villous atrophy and strongly increased intraepithelial CD103+ CD8+ T cells in the small intestine which harbor an increased risk for T-cell lymphoproliferative disorders. Clinical course, histology, and response to immunosuppressive therapy all suggest an autoimmune pathogenesis.

Quantifying intraepithelial lymphocytes and subepithelial collagen band in microscopic colitis, extracting insights into the interrelationship of lymphocytic and collagenous colitis

Microscopic colitis (MC) is the umbrella term for the conditions termed lymphocytic colitis (LC) and collagenous colitis (CC). LC with thickening of the subepithelial collagen band or CC with increased number of intraepithelial T- lymphocytes (IELs) is often seen in MC and may lead to difficulties in correct histological classification.We investigated the extent of overlapping features of CC and LC in 60 cases of MC by measuring the exact thickness of the subepithelial collagen band in Van Gieson stained slides and quantifying number of IELs in CD3 stained slides by digital image analysis. A thickened collagen band was observed in nine out of 29 cases with LC (31%) and an increased number of IELs in all 23 cases of CC (100%). There was no correlation between the thickness of the collagen band and number of IELs.Due to the increased number of IELs in all cases of CC we consider the lymphocytic inflammatory infiltration of the mucosa to be the essential histopathological feature of MC. However, although LC and CC are related due to the lymphocytic inflammation, the non-linear correlation of number of IELs and thickness of the collagenous band indicate differences in their pathogenesis.

Pembrolizumab in unresectable or metastatic MSI-high colorectal cancer: safety and efficacy

ABSTRACT Introduction Microsatellite instability-high (MSI-H) colorectal cancer (CRC) represents a unique subset of CRC characterized by elevated neoantigen expression and a high degree of intraepithelial T-cell infiltrate. These characteristics make MSI-H tumors particularly susceptible to immune checkpoint inhibitors (ICIs) such as pembrolizumab which inhibit the negative regulation of cytotoxic T-cells and promote T-cell mediated anti-tumor activity. Areas covered We discuss the drug development of pembrolizumab including the seminal studies which enabled the drug to garner FDA approvals in the refractory and first-line settings for patients with MSI-H CRC, the pharmacokinetic & pharmacodynamic profile of the agent, and the adverse event profile of the ICI. We also discuss unmet needs in the arena of ICIs including strategies to overcome tumor resistance and to increase the applicability of the agents to a broader population of CRC patients. Expert opinion Despite the anti-tumor activity of pembrolizumab in patients with MSI-H CRC, 30–35% of patients fail to derive any benefit. Ongoing research efforts are seeking to identify ICI combinations, which can overcome CRC resistance to pembrolizumab, move ICIs into the treatment paradigm for patients with localized MSI-H CRC and enable ICIs to become meaningful treatment options for patients with microsatellite stable CRC.

A New Intraepithelial \u03b3\u03b4 T-Lymphocyte Marker for Celiac Disease Classification in Formalin-Fixed Paraffin-Embedded (FFPE) Duodenal Biopsies

BackgroundThe histopathologic diagnosis of celiac disease (CD) may be challenging when the duodenal biopsies mucosal injury is limited. Intraepithelial T-lymphocytes (IELs) can be useful to characterize the degree of mucosal inflammation. A small fraction of IELs expresses the γδ T-cell receptor (named γδ-IELs), whose density, determined by flow cytometry or frozen section immunohistochemistry (IHC), is a specific marker for CD.AimTo establish a new IHC assay for γδ-IELs applicable to formalin-fixed paraffin-embedded (FFPE) duodenal biopsies.MethodsWe analyzed γδ-IELs using IHC in 138 duodenal biopsies using a standard IHC staining protocol with a new monoclonal antibody H-41. IELs were quantitated with digital image analysis.ResultsCompared to those in non-celiac controls (n = 51), γδ-IEL density was significantly increased in newly diagnosed celiac disease patients (n = 22, p < 0.0001). In ROC-curve analysis, the cutoff of 6.5 γδ-IELs/100 enterocytes distinguished optimally active CD patients from non-celiac controls (sensitivity 96%, specificity 95%). γδ-IEL density in CD patients on a gluten-free diet (n = 53) were also higher than in controls (p < 0.0001), but lower than those in newly diagnosed CD (p < 0.0001). The diagnostic value of γδ-IELs outperformed that of CD3 + IELs in both patient groups. γδ-IELs were better than CD3 + IELs distinguishing between celiac disease and conditions histologically mimicking celiac disease (n = 12).ConclusionsIntraepithelial γδ T-lymphocytes can be stained and quantitated reliably in FFPE duodenal biopsies. The results showed excellent specificity and sensitivity for celiac disease. The new IHC method of detection of γδ-IELs is a promising addition to the routine histopathologic assessment methodology of celiac disease.

HISTOPATHOLOGY OF CELIAC DISEASE. POSITION STATEMENTS OF THE ITALIAN GROUP OF GASTROINTESTINAL PATHOLOGISTS (GIPAD-SIAPEC)

Celiac Disease (CeD) is an immune-mediated inflammatory disorder of the small intestine, affecting genetically susceptible individuals when exposed to gluten. Small intestinal biopsy interpretation has been the “gold standard” for celiac disease (CeD) for over 50 years. Despite today’s availability of sensitive and specific serological tests, the histopathological features from mucosal biopsy play a key role in diagnosing when CeD is suspected. Such a diagnostic approach requires a multidisciplinary team to optimize both tissue sampling and interpretation via the interaction between the pathologist and the gastroenterologist. Pathologists of the Italian Group of Gastrointestinal Pathology (GIPAD-SIAPEC), together with a member (TR) of the Italian Society of Technicians (AITIC) and an expert gastroenterologist (CC), provide position statements as a practical tool for reading and interpreting the report.Moreover, a position statement was formulated about the recently described condition known as Non-Celiac Gluten Sensitivity (NCGS). Within such a diagnostic setting, both the architectural abnormalities of the duodenal mucosa, namely glandular hyperplasia, and villous atrophy and the number of intraepithelial T-lymphocytes should be well highlighted. Ancillary tests such as anti-CD3 stain are useful for an accurate count of the intraepithelial T lymphocytes when CeD or NCGS is suspected. Moreover, anti-CD3 and anti-CD8 stains are recommended in patients not responding to the gluten-free diet (GFD) to confirm a diagnosis of Refractory Celiac Disease (RCeD). Diagnostic clues about the differential diagnosis of both CeD and RCeD have also been rendered.

Coffee Intake and Colorectal Cancer Incidence According to T-Cell Response.

We hypothesized that the associations between coffee intake and colorectal cancer (CRC) incidence might differ by immune cell densities in CRC tissue. Using the Nurses’ Health Study and the Health Professionals Follow-up Study, we examined the association of coffee intake with incidence of CRC classified by intraepithelial or stromal T-cell subset densities by multiplex immunofluorescence assay for CD3, CD4, CD8, CD45RO (PTPRC), and FOXP3. We applied an inverse probability-weighted Cox proportional hazardsregression model to control for selection bias and potential confounders. During follow-up of 133 924 participants (3 585 019 person-years), we documented 3161 incident CRC cases, including 908 CRC cases with available data on T-cell densities in tumor tissue. The association between coffee intake and CRC was not statistically significantly different by intraepithelial or stroma T-cell subset (Pheterogeneity > .38). Hence, there is no sufficient evidence for differential effect of coffee intake on incidence of CRC subtypes classified by T-cell infiltrates.

Cytoprotective effect of Nigella sativa seed on 4-nonylphenol-induced renal damage in the African catfish (Clarias gariepinus)

4-Nonylphenol (4-NP) is a nephrotoxic substance that is highly prevalent in aquatic environments. Nigella sativa seed (NSS) has many biological activities and is widely used throughout the world as a medicinal product. Therefore, in the present study, we investigated the cytoprotective effect of NSS on 4-NP-induced renal damage in African catfish (Clarias gariepinus). Thirty fish were divided into five equal groups: an untreated control group and four groups that were challenged with 4-NP at a dose of 0.1 mg L−1 of aquarium water and fed a basal diet supplemented with 0%, 1%, 2.5%, and 5% NSS, respectively, for 3 weeks. Histological, histochemical, and ultrastructural features of the kidney were then assessed as biomarkers for renal tissue damage. Our results confirmed that 4-NP was a potent cytotoxic agent for the kidney tissue and induced renal damage, with 4-NP-intoxicated fish showing necrosis in the epithelial cells of the renal corpuscles, renal proximal convoluted tubules, and intertubular hematopoietic tissue, as well as loss of or a decrease in microvilli, a decrease in mitochondria, and an increase in the lysosomes in the epithelial cells of the proximal convoluted tubules. The kidneys of 4-NP-intoxicated fish also showed increased numbers of Perls’ Prussian blue-positive melanomacrophage centers and intraepithelial T-lymphocytes in the proximal convoluted tubules and plasma cells. The administration of NSS to 4-NP-challenged fish significantly minimized the cytotoxic effect of 4-NP, maintaining the normal kidney structure, with concentrations of 2.5% and 5% of feed being most effective for protecting the kidney against 4-NP-induced renal damage.

Effect of dietary supplementation with yeast cell wall extracts on performance and gut response in broiler chickens

BackgroundThe dietary supplementation of yeast cell wall extracts (YCW) has been found to reduce pathogenic bacteria load, promote immunoglobulin production, prevent diseases by pro-inflammatory responses, and alter gut microbiota composition. This study evaluated growth and slaughter results, health, gut morphology, immune status and gut transcriptome of 576 male chickens fed two diets, i.e. C (control) or Y (with 250–500 g/t of YCW fractions according to the growth period). At 21 and 42 d the jejunum of 12 chickens per diet were sampled and stained with hematoxylin/eosin for morphometric evaluation, with Alcian-PAS for goblet cells, and antibodies against CD3+ intraepithelial T-cells and CD45+ intraepithelial leukocytes. The jejunum sampled at 42 d were also used for whole-transcriptome profiling.ResultsDietary YCW supplementation did not affect final live weight, whereas it decreased feed intake (114 to 111 g/d; P ≤ 0.10) and improved feed conversion (1.74 to 1.70; P ≤ 0.01). Regarding the gut, YCW supplementation tended to increase villi height (P = 0.07); it also increased the number of goblet cells and reduced the density of CD45+ cells compared to diet C (P < 0.001). In the gut transcriptome, four genes were expressed more in broilers fed diet Y compared to diet C, i.e. cytochrome P450, family 2, subfamily C, polypeptide 23b (CYP2C23B), tetratricopeptide repeat domain 9 (TTC9), basic helix-loop-helix family member e41 (BHLHE41), and the metalloreductase STEAP4. Only one gene set (HES_PATHWAY) was significantly enriched among the transcripts more expressed in broilers fed diet Y. However, a total of 41 gene sets were significantly over-represented among genes up-regulated in control broilers. Notably, several enriched gene sets are implicated in immune functions and related to NF-κB signaling, apoptosis, and interferon signals.ConclusionsThe dietary YCW supplementation improved broiler growth performance, increased gut glycoconjugate secretion and reduced the inflammatory status together with differences in the gut transcriptome, which can be considered useful to improve animal welfare and health under the challenging conditions of intensive rearing systems in broiler chickens.

Mucosal Profiling of Pediatric-Onset Colitis and IBD Reveals Common Pathogenics and Therapeutic Pathways

Pediatric-onset colitis and inflammatory bowel disease (IBD) have significant effects on the growth ofinfants and children, but the etiopathogenesis underlying disease subtypes remains incompletely understood. Here, we report single-cell clustering, immune phenotyping, and risk gene analysis for children with undifferentiated colitis, Crohn's disease, and ulcerative colitis. We demonstrate disease-specific characteristics, as well as common pathogenesis marked by impaired cyclic AMP (cAMP)-response signaling. Specifically, infiltration of PDE4B- and TNF-expressing macrophages, decreased abundance of CD39-expressing intraepithelial Tcells, and platelet aggregation andrelease of 5-hydroxytryptamine at the colonic mucosae were common in colitis and IBD patients. Targeting these pathways by using the phosphodiesterase inhibitor dipyridamole restored immune homeostasis and improved colitis symptoms in a pilot study. In summary, comprehensive analysis of the colonic mucosae has uncovered common pathogenesis and therapeutic targets for children with colitis and IBD.