Goblet Cell Mucus Research Articles

PTPRH Alleviates Airway Obstruction and Th2 Inflammation in Asthma as a Protective Factor.

PurposePTPRH inhibits EGFR activity directly in cancer patients and activated EGFR induces goblet cell hyperplasia and mucus hypersecretion in asthma. However, the function of PTPRH in asthma remains unknown. The purpose of this study was to access the association of PTPRH with asthma and its underlying mechanism.Patients and MethodsWe examined the PTPRH level in asthma patients (n = 108) and healthy controls (n = 35), and analyzed the correlations between PTPRH and asthma-related indicators. Human bronchial epithelial cell (HBECs) transfected with PTPRH and asthma mouse model were set up to investigate the function of PTPRH.ResultsThe expression of PTPRH was significantly increased and correlated with pulmonary function parameters, including airway obstruction, and T-helper2 (Th2) associated markers in asthma patients. PTPRH increased in the house dust mite (HDM)-induced asthmatic mice, while Th2 airway inflammation and Muc5ac suppressed when treated with PTPRH. Accordingly, PTPRH expression was markedly increased in IL-13-stimulated HBECs but PTPRH over-expression suppressed MUC5AC. Moreover, HBECs transfected with over-expressed PTPRH inhibited the phosphorylation of EGFR, ERK1/2 and AKT, while induced against PTPRH in HBECs dephosphorylated of EGFR, ERK1/2 and AKT.ConclusionPTPRH reduces MUC5AC secretion to alleviate airway obstruction in asthma via potential phosphorylating of EGFR/ERK1/2/AKT signaling pathway, which may provide possible therapeutic implications for asthma.

Giardia duodenalis cysteine proteases cleave proteinase-activated receptor-2 to regulate intestinal goblet cell mucin gene expression

Giardia duodenalis cysteine proteases have been identified as key virulence factors and have been implicated in alterations to intestinal goblet cell activity and mucus production during Giardia infection. The present findings demonstrate a novel mechanism by which Giardia cysteine proteases modulate goblet cell activity via cleavage and activation of protease-activated receptor 2. Giardia duodenalis (assemblage A) increased MUC2 mucin gene expression in human colonic epithelial cells in a manner dependent upon both protease-activated receptor 2 activation and Giardia cysteine protease activity. Protease-activated receptor 2 cleavage within the N-terminal activation domain by Giardia proteases was confirmed using a nano-luciferase tagged recombinant protease-activated receptor 2. In keeping with these observations, the synthetic protease-activated receptor 2-activating peptide 2fLIGRLO-amide increased Muc2 gene expression in a time-dependent manner. Calcium chelation and inhibition of the ERK1/2 mitogen activated protein kinase pathway inhibited Muc2 upregulation during Giardia infection, consistent with canonical protease-activated receptor 2 signaling pathways. Giardia cysteine proteases cleaved both recombinant protease-activated receptor 1 and protease-activated receptor 2 within their extracellular activation domains with isolate-dependent efficiency that correlated with the production of cysteine protease activity. Protease-activated receptors represent a novel target for Giardia cysteine proteases, and these findings demonstrate that protease-activated receptor 2 can regulate mucin gene expression in intestinal goblet cells.

Nigella sativa oil protects against cadmium-induced intestinal toxicity via promotion of anti-inflammatory mechanisms, mucin expression and microbiota integrity.

Objective:This study examined the protective effects of Nigella sativa oil (NSO) on cadmium (Cd)-induced alterations affecting gut morphology and microbiota composition, as well as the involvement of mucus glycoprotein (MUC2) and immuno-inflammatory markers (TNFα and IL-2) in the colon of rats.Materials and Methods:Male Wistar rats, randomized into four groups, were treated either with distilled water (control), CdCl2 (100 mg/kg), CdCl2+NSO (1 ml/kg) or NSO alone. After the experiments, faecal samples were processed for microbial culture on various selective media, while intestinal segments were prepared for histopathological examination and immunohistochemistry. The composition of NSO was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS).Results:Oral Cd administration provoked dramatic increases in faecal counts of potentially pathogenic bacteria (Staphylococci, Enterococci, Pseudomonas and Escherichia coli), while decreasing probiotic lactobacilli counts. Cadmium treatment caused down-regulation of colonic MUC2 (p=0.003) and IL-2 (p=0.03), but increased TNFα (p=0.034), along with reduced goblet cell counts and mucus production. Conversely, treatment with NSO significantly improved Lactobacilli counts (p=0.042), while reducing the levels of potentially pathogenic species. In addition, NSO significantly restored colonic expressions of MUC2 (p=0.001), TNFα (p=0.007) and IL-2 (p=0.025) to control levels. GC-MS analysis of NSO revealed the presence of the active ingredient, thymoquinone and a high content of unsaturated fatty acids, including trans-13-octadecenoic acid and oleic acid. Conclusion:This study highlights the intestinal mucus, microbiota and immuno-inflammatory system as important protective targets of NSO against Cd-induced intestinal toxicity.

Effects of a polyherbal formulation on inflammation and histopathological alterations in mice with ovalbumin-induced allergic asthma.

Objective:Allergic asthma is a complex inflammatory disorder that affects the airways. As an ancient medical system, Iranian Traditional Medicine (ITM) recommends a polyherbal formula called “Monzej-e-balgham” for the treatment of asthma. In the present investigation, the antiasthmatic effects of “Monzej-e-balgham” were examined in a murine model of allergic asthma.Materials and Methods:Twenty-eight Balb/c mice weighing 15-20 g were allocated into 4 groups. As negative and positive controls, groups I and II received phosphate-buffered saline (PBS) and ovalbumin (OVA) solutions, respectively. Groups III and IV were first sensitized with OVA and then respectively treated with “Monzej-e-balgham” (63 mg/kg) and budesonide. Finally, bronchoalveolar lavage fluid (BALF) and lung tissues of the animals were collected and used for eosinophil counting, Th2 type interleukins (IL-5, IL-13, and IL-33) measurement, and histological examinations.Results:“Monzej-e-balgham” significantly reduced the number of eosinophils and the levels of IL-5, IL-13, and IL-33 in BALF specimens compared to OVA-sensitized group (p<0.05). It also ameliorated histopathological changes of the lung tissues such as goblet cells hyperplasia and mucus overproduction in comparison to group II. Interestingly, the results of the “Monzej-e-balgham”-treated group were comparable with those obtained for budesonide-inhaled mice.Conclusion:The present data indicated a mechanism that involves Th2 inflammatory responses in allergic asthma and suggested a polyherbal mixture for the treatment of this disease.

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.

Maternal gut microbiome regulates immunity to RSV infection in offspring.

Development of the immune system can be influenced by diverse extrinsic and intrinsic factors that influence the risk of disease. Severe early life respiratory syncytial virus (RSV) infection is associated with persistent immune alterations. Previously, our group had shown that adult mice orally supplemented with Lactobacillus johnsonii exhibited decreased airway immunopathology following RSV infection. Here, we demonstrate that offspring of mice supplemented with L. johnsonii exhibit reduced airway mucus and Th2 cell-mediated response to RSV infection. Maternal supplementation resulted in a consistent gut microbiome in mothers and their offspring. Importantly, supplemented maternal plasma and breastmilk, and offspring plasma, exhibited decreased inflammatory metabolites. Cross-fostering studies showed that prenatal Lactobacillus exposure led to decreased Th2 cytokines and lung inflammation following RSV infection, while postnatal Lactobacillus exposure diminished goblet cell hypertrophy and mucus production in the lung in response to airway infection. These studies demonstrate that Lactobacillus modulation of the maternal microbiome and associated metabolic reprogramming enhance airway protection against RSV in neonates.

Effects of cigarette smoke on Haemophilus influenzae-induced otitis media in a rat model

Exposure to cigarette smoke (CS) is a factor that could delay or worsen the recovery of otitis media (OM) by causing inflammatory swelling of the Eustachian tube (ET). However, despite the suggested relationship, little is known about the association between OM and CS. Therefore, we aimed to evaluate the effects of CS on the development, progression, and recovery of OM, as well as the histological and molecular changes caused by CS exposure, by using a rat model of OM infected with non-typeable Haemophilus influenzae (NTHi). Eighty Sprague–Dawley rats with normal middle ears (MEs) were divided into four groups (n = 20 rats/group): control, CS, OM, and CS + OM. The CS and CS + OM groups were exposed to CS for 2 weeks. The inflammatory reaction to NTHi was more intense and lasted longer in the CS + OM group than in the other groups. Goblet cell proliferation and mucus secretion in the ET were more significant in the CS and CS + OM groups than in the other groups. These findings suggested that because CS directly affects the ET and ME mucosa, bacterial OM can become more severe and may resolve more slowly in the presence of CS exposure rather than in its absence.

Mucus Release and Airway Constriction by TMEM16A May Worsen Pathology in Inflammatory Lung Disease.

Activation of the Ca2+ activated Cl− channel TMEM16A is proposed as a treatment in inflammatory airway disease. It is assumed that activation of TMEM16A will induce electrolyte secretion, and thus reduce airway mucus plugging and improve mucociliary clearance. A benefit of activation of TMEM16A was shown in vitro and in studies in sheep, but others reported an increase in mucus production and airway contraction by activation of TMEM16A. We analyzed expression of TMEM16A in healthy and inflamed human and mouse airways and examined the consequences of activation or inhibition of TMEM16A in asthmatic mice. TMEM16A was found to be upregulated in the lungs of patients with asthma or cystic fibrosis, as well as in the airways of asthmatic mice. Activation or potentiation of TMEM16A by the compounds Eact or brevenal, respectively, induced acute mucus release from airway goblet cells and induced bronchoconstriction in mice in vivo. In contrast, niclosamide, an inhibitor of TMEM16A, blocked mucus production and mucus secretion in vivo and in vitro. Treatment of airway epithelial cells with niclosamide strongly inhibited expression of the essential transcription factor of Th2-dependent inflammation and goblet cell differentiation, SAM pointed domain-containing ETS-like factor (SPDEF). Activation of TMEM16A in people with inflammatory airway diseases is likely to induce mucus secretion along with airway constriction. In contrast, inhibitors of TMEM16A may suppress pulmonary Th2 inflammation, goblet cell metaplasia, mucus production, and bronchoconstriction, partially by inhibiting expression of SPDEF.

The role of airway remodeling in the pathogenesis and treatment of chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is currently considered the third leading cause of death in the world. COPD represents an important public health challenge and a socio-economical problem that is preventable and treatable. The main cause of COPD is chronic inhalation of cigarette smoke, and other harmful constituents of air pollution, which cause epithelial injury, chronic inflammation and airway remodeling. Airway remodeling is most prominent in small airways. It is due to infiltration of the airways by inflammatory cells, such as neutrophils, eosinophils, macrophages, and immune cells, including CD8+ T-cells, Th1, Th17 lymphocytes, and innate lymphoid cells group 3. Fibroblasts, myofibroblasts, and airway smooth muscle (ASM) cells also contribute to airway remodeling by depositing extracellular matrix (ECM) proteins, which increase the thickness of the airway wall. Activated inflammatory cells, and structural cells secrete cytokines, chemokines, growth factors, and enzymes which propagate airway remodeling. Airway remodeling is an active process which leads to thickness of the reticular basement membrane, subepithelial fibrosis, peribronchiolar fibrosis, and ASM cells hyperplasia and hypertrophy. It is also accompanied by submucosal glands and goblet cells hypertrophy and mucus hypersecretion, and angiogenesis. Epithelial mesenchymal transmission (EMT) plays a key role in airway remodeling. In patients with COPD and smokers, cellular reprograming in epithelial cells leads to EMT, whereby epithelial cells assume a mesencymal phenotype. Additionally, COPD is associated with increased parasympathetic cholinergic activity, which leads to ASM cells hypercontractility, increased mucus secretion, and vasodilatation. Treatment of COPD is intricate because of the heterogeneous nature of the disease, which requires specific treatment of the pathophysiological pathways, such as airway inflammation, ASM cell hypercontractility, and parasympathetic cholinergic hyperreactivity. The Global Initiative for Chronic Obstructive Lung Disease (GOLD) 2020 strategy report recommends personalized approach for the treatment of COPD. However, some patients with COPD are unresponsive to the standards of care. They may require a triple combination of LABA/LAMA/ICS. Single-inhaler triple therapy (SITT), such as fluticasone fuorate/vilanterol/umeclidinium has been shown to significantly improve symptoms and asthma control, reduce moderate and severe exacerbations, and to improve lung function.

Foxo1 controls gut homeostasis and commensalism by regulating mucus secretion.

Mucus produced by goblet cells in the gastrointestinal tract forms a biological barrier that protects the intestine from invasion by commensals and pathogens. However, the host-derived regulatory network that controls mucus secretion and thereby changes gut microbiota has not been well studied. Here, we identify that Forkhead box protein O1 (Foxo1) regulates mucus secretion by goblet cells and determines intestinal homeostasis. Loss of Foxo1 in intestinal epithelial cells (IECs) results in defects in goblet cell autophagy and mucus secretion, leading to an impaired gut microenvironment and dysbiosis. Subsequently, due to changes in microbiota and disruption in microbiome metabolites of short-chain fatty acids, Foxo1 deficiency results in altered organization of tight junction proteins and enhanced susceptibility to intestinal inflammation. Our study demonstrates that Foxo1 is crucial for IECs to establish commensalism and maintain intestinal barrier integrity by regulating goblet cell function.

17,18-Epoxyeicosatetraenoic Acid Inhibits TNF-α-Induced Inflammation in Cultured Human Airway Epithelium and LPS-Induced Murine Airway Inflammation.

17,18-Epoxyeicosatetraenoic acid (17,18-EpETE), an eicosapentaenoic acid metabolite, is generated from dietary oil in the gut, and antiinflammatory activity of 17,18-EpETE was recently reported. To evaluate the inhibitory effects of 17,18-EpETE in airway inflammation, we examined in vitro and in vivo effects on mucus production, neutrophil infiltration, and cytokine/chemokine production in airway epithelium. Nasal tissue localization of G protein-coupled receptor 40 (GPR40), a receptor of 17,18-EpETE, was determined by immunohistochemical staining. Expression of GPR40 mRNA in nasal mucosa of chronic rhinosinusitis (CRS) patients and control subjects was determined by reverse transcription-polymerase chain reaction (RT-PCR). The in vitro effects on airway epithelial cells were examined using normal human bronchial epithelial cells and NCI-H292 cells. To examine the in vivo effects of 17,18-EpETE on airway inflammation, we induced goblet cell metaplasia, mucus production, and neutrophil infiltration in mouse nasal epithelium by intranasal lipopolysaccharide (LPS) instillation. GPR40 is mainly expressed in human nasal epithelial cells and submucosal gland cells. RT-PCR analysis revealed that the expression of GPR40 mRNA was increased in nasal tissues from CRS patients compared with those from control subjects. 17,18-EpETE significantly inhibited tumor necrosis factor (TNF)-α-induced production of interleukin (IL)-6 , IL-8, and mucin from cultured human airway epithelial cells dose dependently, and these antiinflammatory effects on cytokine production were abolished by GW1100, a selective GPR40 antagonist. Intraperitoneal injection or intranasal instillation of 17,18-EpETE significantly attenuated LPS-induced mucus production and neutrophil infiltration in mouse nasal epithelium. Inflammatory cytokine/chemokine production in lung tissues and bronchoalveolar lavage fluids was also inhibited. These results indicate that 17,18-EpETE plays a regulatory role in mucus hypersecretion and neutrophil infiltration in nasal inflammation. Local or systemic administration may provide a new therapeutic approach for the treatment of intractable airway disease such as CRS.

Effect of yeast cell wall supplementation on intestinal integrity, digestive enzyme activity and immune traits of broilers

ABSTRACT 1. The protective layer formed by intestinal epithelial cells acts as a barrier preventing the adhesion of pathogenic bacteria, aids digestion and passage of nutrients and reduces damage caused from toxins on the gastrointestinal tract. This study was conducted to investigate the effects of a yeast cell wall-based product (YCW), on broiler intestinal integrity, digestive enzyme capacity and immune function. 2. A 35-d trial involving 246, one-d-of-hatch male broiler chickens was carried out at a trial facility at Agri-Food Biosciences Institute (AFBI, Belfast, UK). Birds were randomly allocated into 6 pens at day of hatch (41 birds/pen; 123 birds/group). Pens were divided into two groups: (1) basal diet and (2) basal diet that incorporated YCW at the manufacturers’ recommended inclusion levels (Alltech Inc., Lexington, Kentucky, USA). 3. In this study, YCW supplementation affected broiler intestinal morphology resulting in greater crypt depth, villus height and surface area, goblet cell density and mucus layer thickness and lower muscularis mucosae thickness. The digestive enzymes, maltase, sucrase and alkaline phosphatase, were significantly higher in the YCW supplemented group compared to the control. The expression levels of pro-inflammatory cytokines, IL-1β, IL-12 and IL-18, were significantly lower as was necroptotic cell death in YCW supplemented birds. 4. In conclusion, under the conditions of this study, YCW supplementation positively affected intestinal health parameters in broilers following 35-d supplementation.

Inverted Ductal Papilloma of the Salivary Gland

This is a case consult of slides stated to be from an excision of a buccal mucosa mass in a 58-year-old-man. The specimen was described as a 3 cm diameter roughly oval tan-gray tissue with a 2 x 1.5 cm mucosal ellipse on the surface that has a central ulcerated punctum. Cut section showed an underlying 1.7 cm diameter roughly oval well-circumscribed mass with a granular tan surface. Histological sections show a papillary lesion with an orifice on the mucosal surface and with epithelial nests invaginating into the underlying lamina propria in a non-infiltrative pattern. (Figure 1) The lesion is composed of papillary epithelial fronds with cleft-like spaces between the fronds. (Figure 2) The papillary fronds are lined by non-keratinizing basaloid stratified squamous cells with a superficial layer of columnar glandular cells along with mucous goblet cells interspersed among the squamous cells. (Figure 3) All the cellular components are devoid of cytologic atypia and mitoses. Based on these microscopic features we signed the case out as inverted ductal papilloma (IDP).&#x0D; &#x0D; Ductal papillomas are uncommon benign epithelial tumors with a papillary configuration that originate from the excretory ductal system of salivary gland acini.1-3 The World Health Organization recognizes two sub-types depending on the growth pattern: an intraductal papilloma (IP) and an IDP.1 An IDP usually presents as an asymptomatic submucosal nodule, measuring about 1.5 centimeters in diameter, and most commonly involving the buccal mucosa, followed by the lips, palate, and floor of the mouth.2,3 Histological sections typically show an unencapsulated though well-circumscribed epithelial proliferation with a papillary configuration on the luminal surface, and a nodular, endophytic or invaginating (“inverted”) configuration at its interface with the underlying lamina propria.2 Both the papillary and the invaginating areas are composed of basaloid, non-keratinizing stratified squamous epithelium that are often covered with a cuboidal or columnar ductal cell layer.2 Scattered among these are mucous goblet cells which can form microcysts.1,2 There is an overall morphological similarity to the sinonasal inverted papilloma.3 A relationship to trauma has been proposed.1,4 Association with Human Papilloma Virus (HPV) has also been reported.1 Others, however, have not been able to demonstrate this association.4&#x0D; &#x0D; Differential diagnoses primarily include IP - which is differentiated from IDP architecturally by being a unicystic intraluminal papillary proliferation within a dilated excretory duct 2 – and sialadenoma papilliferum – which is predominantly polypoid and pedunculated with a verrucoid surface rather than a submucosal nodule, and an over-all morphologic similarity to the cutaneous tumor syringocystadenoma papilliferum.1,4 An important differential diagnosis that has to be ruled out is mucoepidermoid carcinoma (MECA) because of the presence of both squamous and mucin-secreting cells. MECA is distinguished by poor circumscription, and an infiltrative solid-cystic growth pattern.2,4&#x0D; &#x0D; IDP is benign and non-recurrent. Unlike the nasal tumor, there has been no report of malignant transformation.2,3 Complete surgical excision is considered curative.1,2 Reporting these cases is encouraged to further our knowledge of the entity and elucidate a potential association with HPV.

VEGF receptor 2 (KDR) protects airways from mucus metaplasia through a Sox9-dependent pathway.

Mucus-secreting goblet cells are the dominant cell type in pulmonary diseases, e.g., asthma and cystic fibrosis (CF), leading to pathologic mucus metaplasia and airway obstruction. Cytokines including IL-13 are the major players in the transdifferentiation of club cells into goblet cells. Unexpectedly, we have uncovered a previously undescribed pathway promoting mucous metaplasia that involves VEGFa and its receptor KDR. Single-cell RNA sequencing analysis coupled with genetic mouse modeling demonstrates that loss of epithelial VEGFa, KDR, or MEK/ERK kinase promotes excessive club-to-goblet transdifferentiation during development and regeneration. Sox9 is required for goblet cell differentiation following Kdr inhibition in both mouse and human club cells. Significantly, airway mucous metaplasia in asthmatic and CF patients is also associated with reduced KDR signaling and increased SOX9 expression. Together, these findings reveal an unexpected role for VEGFa/KDR signaling in the defense against mucous metaplasia, offering a potential therapeutic target for this common airway pathology.

G Protein-Coupled Receptor Kinase 2 (GRK2) Regulates T Cell Response in a Murine Model of House Dust Mite-Induced Asthma.

G protein-coupled receptor kinase 2 (GRK2) is an adapter protein that modulates G protein-coupled receptor (GPCR) signaling. It also regulates the functions and activity of other intracellular proteins in many cell types. Accordingly, GRK2 is thought to contribute to disease progression by a variety of mechanisms related to its multifunctional roles. Indeed, GRK2 levels are enhanced in patient samples as well as in preclinical models of several diseases. We have previously shown that GRK2 regulates mast cell functions, and thereby contributes to exacerbated inflammation during allergic reactions. In the current study, we observed that GRK2 levels are enhanced in the lungs of human asthma patients and in mice sensitized to house dust mite extract (HDME) allergen. Consistent with these findings, interleukin (IL)-4 and IL-13 levels were reduced in the lungs of GRK2+/− mice in a HMDE mouse model of asthma. Because Th2 cells are the major source of these cytokines during asthma, we determined the role of GRK2 in regulating T cell-specific responses in our HMDE mouse model. We observed a significant reduction of airway hyperresponsiveness (AHR), lung eosinophil and lymphocyte counts, serum IgE, Th2 cytokines (IL-4 and IL-13), goblet cell hyperplasia and mucus production in mice that had reduced GRK2 expression specifically in T cells. Collectively, our studies reveal an important role for GRK2 in regulating T cell response during asthma pathogenesis and further elucidation of the mechanisms through which GRK2 modulates airway inflammation will lead to the development of new therapeutic strategies for asthma.

G-protein coupled receptor kinase 2 (GRK2) regulates T cell response in a murine model of house dust mite-induced asthma

Abstract G protein-coupled receptor kinase 2 (GRK2) is an adapter protein that modulates G protein-coupled receptor (GPCRs) signaling. It also regulates the functions and activity of other intracellular proteins in many cell types. Accordingly, GRK2 is thought to contribute to disease progression by a variety of mechanisms related to its multifunctional roles. Indeed, GRK2 levels are enhanced in patient samples as well as in preclinical models of several diseases. We have previously shown that GRK2 regulates mast cell functions, and thereby contributes to exacerbated inflammation during allergic reactions. In the current study, we observed that GRK2 levels are enhanced in the lungs of human asthma patients and in mice sensitized to house dust mite extract (HDME) allergen. Consistent with these findings, interleukin (IL)-4 and IL-13 levels were reduced in the lungs of GRK2+/− mice in a HMDE mouse model of asthma. Because Th2 cells are the major source of these cytokines during asthma, we determined the role of GRK2 in regulating T cell-specific responses in our HMDE mouse model. We observed a significant reduction of airway hyperresponsiveness (AHR), lung eosinophil and lymphocyte count, serum IgE, Th2 cytokines (IL-4 and IL-13), goblet cell hyperplasia and mucus production in mice that had reduced GRK2 expression specifically in T cells. Collectively, our studies reveal an important role for GRK2 in regulating T cell response during asthma pathogenesis and further elucidation of the mechanisms through which GRK2 modulates airway inflammation will lead to the development of new therapeutic strategies for asthma.

Structural, ultrastructural, and functional aspects of the skin of the upper lip of silver carp (Hypophthalmichthys molitrix).

This study was carried out to analyze the architecture of the skin of the upper lip region in silver carp fishes using light, scanning, and transmission electron microscopies. The skin was composed of epidermis, dermis, and hypodermis. The epidermis of the upper lip was characterized by the presence of large number of metachromatic mucous goblet cells, which showed positive reaction with Periodic Acid-Schiff (PAS), Alcian blue (AB), and toluidine blue. The electroreceptive lateral line system was organized into ampullary and tuberous organs. The scanning electron microscopy showed that the surface of the skin of upper lip was covered by microridges and characterized by the presence of taste buds and openings of lateral line system. As observed by transmission electron microscopy, the cytoplasm of the epidermal layers appeared electron-dense except for the superficial layer, where the cytoplasm was electron-lucent and contained many vacuoles and few profiles of rER. Moreover, the epidermis contained rodlet cells and stem cells. Few organelles were found within the cytoplasm of club cells. Neutrophils and eosinophilic granular cells were also demonstrated as important immune cells in the epidermis of the upper lip. Furthermore, lymphocytes and basophils could be identified with macrophage in the epidermal layer of the upper lip. Numerous telocytes were demonstrated between the collagen fibers of the dermis and bundles of myelinated nerve fibers. In conclusion, the skin of the upper lip region of silver carp displayed many sensory and immunological characteristic features.

ELEVATED ADENOMATOUS POLYPOSIS COLI IN GOBLET CELLS IS ASSOCIATED WITH INFLAMMATION IN MOUSE AND HUMAN COLON

Adenomatous Polyposis Coli (APC) functions as an essential colon tumor suppressor. Roles for APC in other disease states such as Inflammatory Bowel Disease (IBD) remain less defined. In the process of characterizing Apc protein in gastrointestinal tissues from human patients with active IBD, we found a subset of goblet cells with elevated Apc staining intensity. While normal colon tissue was only sparsely populated with these “APChigh” goblet cells, tissue from patients with Ulcerative Colitis and Crohn’s Disease contained entire regions where almost every goblet cell was APChigh. In the descending colon, the APChigh goblet cells were prominent in patients with active Ulcerative Colitis but were mostly lacking in normal samples. In mice, APChigh cells were observed only in the small intestines and proximal/medial colons. Upon induction of colitis with Dextran Sodium Sulfate (DSS), the APChigh goblet cells remained in the proximal and medial colon, but also appeared in the distal colon. To better understand the link between APC expression, goblet cells, and inflammation, cultured human colon cells derived from normal tissue were depleted of APC and RNAseq analysis was performed. APC depletion resulted in reduced expression of mRNAs encoding IL1 signaling pathway components IL1b and IL1R, known regulators of Muc2 expression. Muc2 is the main constituent of goblet cell mucus, which is secreted to form a protective barrier for the epithelial cells lining the large intestine. We found that treating cancer cells lacking wild-type APC with IL1b resulted in increased IL1R and MUC2 expression. Induction of full-length APC in these cells led to a similar phenotype. Combining IL1β treatment with APC induction led to an increase of MUC2 expression greater than expected for additive affects, suggesting that APC sensitized these cells to IL1 signaling. These results illuminate a novel role for APC in goblet cells under conditions of inflammation. We postulate that the higher amount of APC in a subset of goblet cells might sensitize them to IL-1 signaling. During times of inflammation, such as active Ulcerative Colitis, goblet cells with higher APC levels would be more sensitive to IL-1-induced MUC2 expression than other goblet cells, perhaps protecting the intestinal lining from further barrier breaches.

ELEVATED ADENOMATOUS POLYPOSIS COLI IN GOBLET CELLS IS ASSOCIATED WITH INFLAMMATION IN MOUSE AND HUMAN COLON

Abstract Adenomatous Polyposis Coli (APC) functions as an essential colon tumor suppressor. Roles for APC in other disease states such as Inflammatory Bowel Disease (IBD) remain less defined. In the process of characterizing Apc protein in gastrointestinal tissues from human patients with active IBD, we found a subset of goblet cells with elevated Apc staining intensity. While normal colon tissue was only sparsely populated with these “APChigh” goblet cells, tissue from patients with Ulcerative Colitis and Crohn’s Disease contained entire regions where almost every goblet cell was APChigh. In the descending colon, the APChigh goblet cells were prominent in patients with active Ulcerative Colitis but were mostly lacking in normal samples. In mice, APChigh cells were observed only in the small intestines and proximal/medial colons. Upon induction of colitis with Dextran Sodium Sulfate (DSS), the APChigh goblet cells remained in the proximal and medial colon, but also appeared in the distal colon. To better understand the link between APC expression, goblet cells, and inflammation, cultured human colon cells derived from normal tissue were depleted of APC and RNAseq analysis was performed. APC depletion resulted in reduced expression of mRNAs encoding IL1 signaling pathway components IL1b and IL1R, known regulators of Muc2 expression. Muc2 is the main constituent of goblet cell mucus, which is secreted to form a protective barrier for the epithelial cells lining the large intestine. We found that treating cancer cells lacking wild-type APC with IL1b resulted in increased IL1R and MUC2 expression. Induction of full-length APC in these cells led to a similar phenotype. Combining IL1β treatment with APC induction led to an increase of MUC2 expression greater than expected for additive affects, suggesting that APC sensitized these cells to IL1 signaling. These results illuminate a novel role for APC in goblet cells under conditions of inflammation. We postulate that the higher amount of APC in a subset of goblet cells might sensitize them to IL-1 signaling. During times of inflammation, such as active Ulcerative Colitis, goblet cells with higher APC levels would be more sensitive to IL-1-induced MUC2 expression than other goblet cells, perhaps protecting the intestinal lining from further barrier breaches.

Morph-anatomic and histochemical study of ileum of goose (Alopochen egyptiacus) with special references to immune cells, mucous and serous goblet cells, telocytes, and dark and light smooth muscle fibers.

The morphological characteristics of the ileum of 20 adult male Egyptian geese were determined using LM, SEM, and TEM. The mean length of the ileum in the male goose was approximately 158.71 mm, representing nearly 10.19% of the total length of the small intestine. The ileum is composed of four layers: mucosa, submucosa, muscular layer, and serosa. The mucosal layer comprises the epithelium, lamina propria, and muscularis mucosa. The mucosa forms finger-like villi and is invaginated at the bases, forming the crypts of Lieberkühn. The ileum is lined by simple columnar epithelium that contains absorptive dark and light enterocytes with two types of goblet cells (mucous and serous varieties) microfold like cells, dendritic reticulum cells, Paneth cells, and a closed type of enteroendocrine cells. The lamina propria has diffuse lymphoid tissue containing lymphocytes, macrophages, mast cells, plasma cells, and heterophils as well as telocytes. The muscularis mucosa comprises circular smooth muscle fibers extending into the core of the villi. The submucosa is a thin layer of elastic-rich connective tissue. The muscular level consists of four layers, with light and dark smooth muscle fibers. We described in detail the structure of all cellular components and histomorphometric measurements.