Mast cells in oral lichen planus and oral lichenoid lesions related to dental amalgam contact

Abstract The aim of this study was to analyze the expression of mast cell markers toluidine blue, c-kit, and tryptase and presence of mononuclear inflammatory cells in oral lichen planus (OLP) and oral lichenoid lesions related to dental amalgam. Nineteen specimens of OLP, OLLC, and healthy oral mucosa were selected. Mononuclear inflammatory cells were analyzed. Histochemical and immunohistochemical analyses were performed using toluidine blue, anti-c-kit and anti-tryptase reagents, and the results were quantified in areas A and B of connective tissue. Mast cells of all OLP and OLLC samples were positive for toluidine blue, c-kit, and tryptase. The density of toluidine blue+, c-kit+ and tryptase+ mast cells was higher in tissue with OLP and OLLC compared with healthy controls (p < 0.05). No difference was noted in mast cells density between OLP and OLLC (p > 0.05). The density of tryptase+ mast cells was higher in the subepithelial region (area A) than the region below it (Area B) in OLLC (p = 0.047). The mononuclear inflammatory cell density was higher in OLLC compared to OLP, but without statistical significance (p > 0.05). A positive statistical correlation was found between mononuclear immune cells and density of c-kit+ and tryptase+ mast cells in OLP (r = 0.943 and r = 0.886, respectively). Our data demonstrate that the etiopathogenesis process of OLP and OLLC modulates the expansion and degranulation of mast cells; mast cells density, however, was similar between OLP and OLLC. The distribution of mast cells appears to vary along the lamina propria.

Mastócitos e síndrome de ativação mastocitária: o que há de novo? / Mast cells and mast cell activation syndrome: what's new?

Os mastócitos são as principais células efetoras da resposta alérgica aguda, desempenhando também um papel importante na angiogênese, tolerância imunológica, regulação da fibrinólise, regeneração neuronal e osteoclastogênese. Localizam-se maioritariamente na pele e nas mucosas do intestino e pulmões, onde exercem uma função "sentinela". As síndromes de ativação mastocitária são caracterizadas pela ocorrência de episódios recorrentes de manifestações clínicas resultantes da libertação de mediadores mastocitários. Esta constitui-se como entidade complexa com um espectro de sintomas associados, representando um desafio diagnóstico e terapêutico. Nesta revisão, os autores pretendem apresentar uma visão geral sobre a estrutura e função dos mastócitos e sobre os critérios diagnósticos e abordagem terapêutica da síndrome de ativação mastocitária.

Mast cells are the main effector cells of acute allergic response, also playing an important role in angiogenesis, immune tolerance, regulation of fibrinolysis, neuronal regeneration, and osteoclastogenesis. They are generally located in the skin and mucous membranes of the intestines and lungs, where they perform a "sentinel" function. Mast cell activation syndrome is characterized by recurrent clinical manifestations resulting from the release of mast cell mediators. This complex entity, which involves a spectrum of associated symptoms, is a diagnostic and therapeutic challenge. In this article we overview of the structure and function of mast cells, in addition to the diagnostic criteria and therapeutic approaches to mast cell activation syndrome.

Evaluation of peripheral nerve fibers and mast cells in burning mouth syndrome

Abstract Emerging evidence has revealed a cross-talk in the etiopathogenesis of burning mouth syndrome (BMS) related to peripheral nerve fibers (NF) and neuropeptides secreted by mast cells. Here, we investigated the S-100+ density and PGP 9.5+ integrity of peripheral NF and the tryptase+ mast cell density in the oral mucosa of BMS patients and healthy individuals. A total of 23 oral mucosa specimens (12 BMS and 11 controls) were evaluated. The clinical diagnosis of BMS was based on a careful examination, excluding other local and systemic causes. Samples were taken from an incisional biopsy of the tongue mucosa of individuals with symptomatic BMS, while the margins of the non-neoplastic tongue biopsy served as controls of healthy individuals. Immunohistochemistry was performed to determine the density/mm2 of S-100+, PGP 9.5+ peripheral NF, and tryptase+ mast cells. Similar densities of S-100+, PGP 9.5+ peripheral NF, and tryptase+ mast cells were found in cases of BMS, with a median value of 3.70, 0.70, and 29.24/mm2, respectively, and in the control group, with a median value of 2.60, 0.80, and 26.01/mm2, respectively (p > 0.05). Moreover, the relationship between S100+ and PGP 9.5+ peripheral NF was the same in both groups (p = 0.70). This study demonstrated that there were no alterations in the density and integrity of peripheral NF in the tongue of symptomatic BMS patients. However, the sensitization of peripheral NF in this disease may not depend on mast cell density.

Increased tumor-associated mast cells facilitate thyroid cancer progression by inhibiting CD8+ T cell function through galectin-9

As an important component of solid tumors, mast cells show specific phenotypes in various tumor microenvironments. However, the precise mechanism of mast cell accumulation and the phenotypic features of thyroid cancer (TC) remain largely unknown. Here, we found that mast cells were obviously recruited to tumor tissue by TC-derived stem cell factor (SCF). With tumor progression, mast cell levels increased gradually. In addition, intratumoral mast cells expressed higher levels of the immunosuppressive molecule galectin-9, which effectively suppresses CD8+ T-cell antitumor immunity in vitro. Blocking galectin-9 on tumor-infiltrating mast cells reversed the immunosuppression of CD8+ T cells. In conclusion, our data elucidated novel protumorigenic and immunosuppressive roles of mast cells in TC. In addition, our results indicated that blocking mast cells may impede tumor progression and ameliorate the prognosis of TC patients.

Presence of mast cells in the rat pericranium ­ a tissue very sensitive to pain / Presença de mastócitos no pericrânio do rato ­ tecido muito sensível à dor

Objective: To evaluate mast cell presence in the pericranium of Wistar rats. Methods: Five male rats of the Wistar strain were used. The animals were housed under a 12 h light cycle with ad libitum access to food and water and allowed 10 days of acclimatization before tissue sampling. The five rats were anesthetized by intraperitoneal injection of ketamine/xylazine, 10/20 mg/kg. Following aseptic preparation of the head skin, a midline longitudinal incision was made to expose the pericranium. Two samples of the pericranium were taken, one from the right and one from the left. These samples were fixed in 10% buffered formaldehyde for 24 h. After fixation, tissue samples were paraffin-embedded and sectioned at 4 µm. Then, slides were deparaffinized, stained with a concentration of 0.1% toluidine blue for 1 min, and washed with distilled water. Last, slides were photomicrographed under 400x magnification to identify mast cells. Results: Mast cells were identified in the dura mater and the five rats' pericranium. In the dura mater, mast cells were also found in these rats. We found both granulated (intact) and degranulated mast cells. Conclusion: We suggest that future preclinical studies investigating the involvement of dural mast cells and other meningeal cell populations should also include pericranium samples to explore this structure's relevance in migraine pain and other headache disorders.

Objetivo: Avaliar a presença de mastócitos no pericrânio de ratos Wistar. Métodos: Foram utilizados cinco ratos machos da linhagem Wistar. Os animais foram alojados sob um ciclo de luz de 12 horas com acesso ad libitum a comida e água e tiveram 10 dias de aclimatação antes da amostragem de tecido. Os cinco ratos foram anestesiados por injeção intraperitoneal de cetamina/xilazina, 10/20 mg/kg. Após preparação asséptica da pele da cabeça, foi feita uma incisão longitudinal na linha média para expor o pericrânio. Foram retiradas duas amostras do pericrânio, uma da direita e outra da esquerda. Essas amostras foram fixadas em formaldeído tamponado a 10% por 24 horas. Após a fixação, as amostras de tecido foram embebidas em parafina e seccionadas a 4 µm. Em seguida, as lâminas foram desparafinizadas, coradas com concentração de azul de toluidina 0,1% por 1 min e lavadas com água destilada. Por fim, as lâminas foram fotomicrografadas com aumento de 400x para identificação de mastócitos. Resultados: Foram identificados mastócitos na dura-máter e no pericrânio dos cinco ratos. Na dura-máter, mastócitos também foram encontrados nesses ratos. Encontramos mastócitos granulados (intactos) e desgranulados. Conclusão: Sugerimos que futuros estudos pré-clínicos que investiguem o envolvimento de mastócitos durais e outras populações de células meníngeas também incluam amostras de pericrânio para explorar a relevância desta estrutura na dor da enxaqueca e em outros distúrbios de cefaleia.

Progress in the treatment of chronic spontaneous urticaria: insights from the changes in the understanding of its pathogenesis / 中华皮肤科杂志

Mast cells, autoantibodies, inflammatory cells, coagulation cascade, complement system and nervous system are all involved in the complex pathogenesis of chronic spontaneous urticaria (CSU) , while mast cells play a pivotal role in it. With deeper understanding of the pathogenesis of CSU, cutting-edge therapeutic methods are gradually being used in clinical practice. Nowadays, pharmacotherapyeutic studies are more focused on accurately modulating the pathological state of mast cells. This review summarizes recent advances in the pathogenesis of and medicines for CSU.

Mastocitosis del adulto: reporte de un caso / Adult mastocytosis: a case report

RESUMEN La mastocitosis es una enfermedad heterogénea, caracterizada por una acumulación de mastocitos en uno o más órganos, siendo el más afectado la piel. Es más frecuente en niños, pero también se presentan casos en los adultos. Hay diferencias significativas entre las formas de presentación en estos grupos etarios, así como también en su evolución y pronóstico. Presentamos el caso clínico de una paciente con mastocitosis cutánea de inicio en la adultez.

ABSTRACT Mastocytosis is a heterogeneous disease, characterized by an accumulation of mast cells in one or more organs. The skin being is the most frecuently affected organ. It is more common in children, but cases also occur in adults. There are significant differences between the forms of presentation in these age groups, as well as in their evolution and prognosis. We report the case of a patient with adult-onset cutaneous mastocytosis.

Role of mast cells in the pathogenesis of allergic contact dermatitis / 中华皮肤科杂志

Allergic contact dermatitis (ACD) is a T cell-mediated inflammatory skin disease induced by contact allergens. The role of mast cells in ACD remains controversial. In the sensitization phase, mast cells play a pro-inflammatory role by releasing inflammatory cytokines to promote recruitment of neutrophils, and affect the sensitization process by stimulating or inhibiting the migration of dendritic cells. During the elicitation phase, mast cells promote non-allergen-specific inflammation, and enhance allergen-specific inflammation in moderate ACD, but inhibit allergen-specific inflammation in severe and chronic ACD. In addition to the IgE-Fcε receptor I-histamine pathway, Mas-related G-protein coupled receptor B2/X2 (MrgprB2/X2) can mediate the activation of mast cells, leading to the release of tryptase to drive non-histaminergic itch, and may serve as a new target for the treatment of ACD.

Effect of electroacupuncture at "Zusanli" (ST 36) on duodenal mast cells, NGF and NTRK1 in rats with functional dyspepsia / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) on duodenal mast cells, nerve growth factor (NGF) and neurotrophic tyrosine kinase receptor type 1 (NTRK1), and to explore the mechanism of electroacupuncture at Zusanli (ST 36) on functional dyspepsia (FD).@*METHODS@#Sixty SPF-grade 10-day-old SD rats were randomly divided into a normal group, a model group, a ketotifen group and an EA group, 15 rats in each group. The FD model was prepared by iodoacetamide combined with rat tail clamping method in the model group, the ketotifen group and the EA group. The rats in the ketotifen group were injected intraperitoneally with ketotifen (1 mg•kg-1•d-1) for 7 days; the rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), with disperse-dense wave, frequency of 2 Hz/50 Hz and intensity of 0.5 mA, 20 min each time, once a day for 14 days. The gastric emptying rate and small intestinal propulsion rate in each group were observed; the morphology of duodenal mucosa was observed by HE staining; the toluidine blue staining was used to observe the number and degranulation of mast cells in duodenal mucosa; the protein and mRNA expressions of NGF, NTRK1 in duodenum were detected by Western blot and real-time PCR; the level of interleukin-1β (IL-1β) in duodenum was measured by ELISA.@*RESULTS@#Compared with the normal group, the gastric emptying rate and small intestinal propulsion rate in the model group were decreased (P<0.01); compared with the model group, the gastric emptying rate and small intestinal propulsion rate in the ketotifen group and the EA group were increased (P<0.01); the small intestinal propulsion rate in the EA group was higher than that in the ketotifen group (P<0.01). In the model group, local defects in duodenal mucosa were observed with a small amount of inflammatory cell infiltration; no obvious abnormality was found in duodenal mucosa of the other groups. Compared with the normal group, the mast cells of duodenal mucosa in the model group were increased significantly with significant degranulation; compared with the model group, the mast cells of duodenal mucosa in the ketotifen group and the EA group were decreased significantly, and the degranulation was not obvious. Compared with the normal group, the protein and mRNA expressions of NGF, NTRK1 as well as the level of IL-1β in duodenum in the model group were increased (P<0.01); compared with the model group, the protein and mRNA expressions of NGF, NTRK1 as well as the levels of IL-1β in duodenum in the ketotifen group and the EA group were decreased (P<0.01, P<0.05); compared with the ketotifen group, the mRNA expression of NGF, as well as the protein and mRNA expressions of NTRK1 in duodenum in the EA group were decreased (P<0.05, P<0.01).@*CONCLUSION@#EA at "Zusanli" (ST 36) could inhibit the activation of duodenal mast cells and regulate the expressions of NGF and its receptor to improve the low-grade inflammatory response of duodenum, resulting in treatment effect on FD.

Lesiones plantares en paciente pediátrico: apropósito de dos casos / Sole of feet lesions in pediatric patients: a purpose of two cases

RESUMEN Las mastocitosis e histiocitosis, son enfermedades que se caracterizan por la proliferación o activación descontrolada y posterior acumulación anormal de mastocitos e histiocitos respectivamente. De incidencia desconocida, talvez porque son subdiagnosticadas. Su patogenia aún es desconocida, si bien está relacionada con mutaciones en la vía del C-KIT para las mastocitosis y de origen viral o neoplásico en el caso de las histiocitosis. Ambas patologías suelen ser frecuentes en la infancia, incluso algunas son congénitas. El mastocitoma cutáneo único sería una forma benigna de mastocitosis y la histiocitosis de células de Langerhans es una forma de histiocitosis que en nuestro caso al afectar un solo órgano (la piel) tendría un buen pronóstico.

SUMMARY Mastocytosis and histiocytosis are diseases that are characterice by uncontrolled proliferation or activation and subsequent abnormal activation of mast cells and histiocytes respectively. Of unknown incidence, perhaps because they are underdiagnosed, their pathogenesis is still unknown although it is related to mutations in the C-KIT pathway for mastocytosis and of viral or neoplastic origin in the case of histiocytosis. Both pathologies are usually frequent in childhood, even some are congenital. The single cutaneous mastocytoma would be a benign form of mastocytosis and the histiocytosis of Langerhans cells is a form of histiocytosis that in our case affecting a single organ (the skin) will have a good prognosis.

Involvement of tryptase-positive mast cells and angiogenesis in the growth of inflammatory odontogenic cysts

Abstract: Inflammatory periapical lesions are characterized by infiltration of different immune cell types, the functions of which depend on an effective vascular network. This study aimed to evaluate the mast cells density (MCD) in inflamatory odontogenic cysts capsules concerning microvascular density (MVD), microvascular area (MVA), and microvascular perimeter (MVP), and correlate such findings with the type of lesion, intensity of the inflammatory infiltrate, and thickness of the epithelial lining. Twenty inflamatory dentigerous cysts (IDCs), twenty radicular cysts (RCs), and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using anti-tryptase and anti-CD34 antibodies. RCs exhibited the highest MCD, MVD, MVA, and MVP indexes (p = < 0.001, p = 0.008, p = 0.003 and p = < 0.001, respectively), and lesions with inflammatory infiltrate grade III showed the highest MVD (p = 0.044). Considering epithelial thickness, a higher MVP index was identified in lesions with hyperplastic epithelium (p = 0.018). In IDCs, RCs, and RRCs, a strong positive correlation was observed between MVA and MVP (r = 0.950 and p = < 0.001; r = 0.914 and p = < 0.001; r = 0.713 and p = < 0.001, respectively). In IDCs, a moderate correlation was observed between MCD and both MVA and MVP (r = 0.660 and p = 0.002; r = 0.634 and p = 0.003, respectively). These results suggest that tryptase-positive mast cells might play an important role in the angiogenic activity of IDCs, while RCs had the highest indexes. Our findings also confirmed that the intensity of the inflammatory infiltrate and epithelial thickness influence angiogenesis.

Profibrotic effect of mast cells on hyperoxia-induced lung injury / 国际儿科学杂志

Bronchopulmonary dysplasia(BPD)is a chronic lung disease that usually occurs in premature infants who require mechanical ventilation, with a high mortality rate, and there is currently a lack of effective prevention and treatment methods.The pathological features of typical BPD are mainly severe airway injury, lung parenchymal inflammation, squamous epithelial metaplasia, and pulmonary fibrosis.Although the pathogenesis of BPD is not yet clear, a large number of mast cell aggregation can be observed in the fibrotic lung of premature infants.Studies have suggested that a series of inflammatory mediators released by mast cell degranulation under hyperoxia have an important impact on BPD.This article reviews the role and mechanism of mast cells in promoting fibrosis in BPD, and in order to provide new ideas for the treatment of BPD.

Diagnosis and treatment of chronic urticaria: current status and future prospects / 中华皮肤科杂志

The etiology and pathogenesis of chronic urticaria are complex. The main traditional treatment is oral antihistamines. With the progressive development in the biomedical field, targeted therapy has gradually become a new treatment option. Anti-immunoglobulin E monoclonal antibodies (omalizumab) can rapidly improve patients′ condition and enhance their quality of life during the treatment of chronic urticaria, and its clinical efficacy and safety have been gradually confirmed in clinical practice. This article summarizes and analyzes the current status of clinical diagnosis and treatment of chronic urticaria, discusses some common problems and corresponding strategies, and provides a reference for clinical management of these patients.

Mast cells in the pathophysiology of Duchenne muscular dystrophy in Golden Retriever dogs / O papel dos mastócitos na patogênese da distrofia muscular de Duchenne em cães Golden Retriever

The Golden Retriever muscular dystrophy (GRMD) is one of the best models of Duchenne muscular dystrophy (DMD), with similar genotypic and phenotypic manifestations. Progressive proliferation of connective tissue in the endomysium of the muscle fibers occurs in parallel with the clinical course of the disease in GRMD animals. Previous studies suggest a relationship between mast cells and the deposition of fibrous tissue due to the release of mediators that recruit fibroblasts. The aim of this study was to evaluate the presence of mast cells and their relationship with muscle injury and fibrosis in GRMD dogs of different ages. Samples of muscle groups from six GRMD and four control dogs, aged 2 to 8 months, were collected and analyzed. The samples were processed and stained with HE, toluidine blue, and Azan trichrome. Our results showed that there was a significant increase in infiltration of mast cells in all muscle groups of GRMD dogs compared to the control group. The average number of mast cells, as well as the deposition of fibrous tissue, decreased with age in GRMD dogs. In the control group, all muscle types showed a significant increase in the amount of collagenous tissue. This suggests increased mast cell degranulation occurred in younger GRMD dogs, resulting in increased interstitial space and fibrous tissue in muscle, which then gradually decreased over time as the dogs aged. However, further studies are needed to clarify the role of mast cells in the pathogenesis of fibrosis.(AU)

O cão Golden Retriever distrófico (Golden Retriever muscular dystrophy - GRMD) é um dos melhores modelos da distrofia muscular de Duchenne (DMD), com manifestações genotípicas e fenotípicas similares. A proliferação progressiva de tecido conjuntivo no endomísio das fibras musculares ocorre paralelamente ao curso clínico da doença em animais GRMD. Estudos anteriores sugerem uma relação entre os mastócitos e a deposição de tecido fibroso devido à liberação de mediadores que recrutam fibroblastos. O objetivo deste estudo foi avaliar a presença de mastócitos e sua relação com a lesão muscular e fibrose em cães GRMD de diferentes idades. Amostras de grupos musculares de seis GRMD e quatro controles, com idade entre 2 a 8 meses, foram coletadas e analisadas. As amostras foram processadas e coradas com HE, azul de toluidina e tricrômico de Azan. Nossos resultados mostraram que houve um aumento significativo na infiltração de mastócitos em todos os grupos musculares de cães GRMD em comparação com o grupo controle. O número médio de mastócitos, assim como a deposição de tecido fibroso, diminuiu com a idade em cães GRMD. No grupo controle, todos os tipos musculares mostraram um aumento significativo na quantidade de tecido colágeno. Isto sugere o aumento da degranulação de mastócitos em cães GRMD mais jovens, resultando em aumento do espaço intersticial e tecido fibroso no músculo, que então gradualmente diminuiu com o tempo à medida que os cães envelheceram. No entanto, mais estudos são necessários para esclarecer o papel dos mastócitos na patogênese da fibrose.(AU)

Mast cells in the kidney biopsies of pediatric patients with lupus nephritis / Mastócitos em biópsias renais de pacientes pediátricos com nefrite lúpica

ABSTRACT Introduction: Mast cells may be involved in inflammation and contribute to the onset of fibrosis in lupus nephritis (LN). Objective: This study aimed to correlate the presence of mast cells in kidney biopsy specimens of pediatric patients with LN with activity (AI) and chronicity (CI) indices and assess how effectively mast cells may be used as a prognostic factor. Method: The study included 40 patients aged 6-18 years diagnosed with LN at the Renal Disease Service of the Federal University of Triângulo Mineiro between 1996 and 2015. Workup and epidemiological data were evaluated vis-à-vis AI, CI, and mast cell counts (MCC). Results: Significant positive correlations were found between mast cell counts (MCC) and AI (p = 0.003; r: 0.66) and MCC and CI (p = 0.048; r: 0.48). The ROC curve showed that mast cells were highly sensitive and specific in the differentiation of patients with an AI > 12 from individuals with an AI ≤ 12. Serum creatinine levels were higher in individuals with class IV LN than in patients with class V disease [1.50 (0.40-20.90) vs. 0.70 (0.62-0.90), p = 0.04]. Blood urea nitrogen had a positive significant correlation with MCC (p = 0.002; r: 0.75). A trend toward a negative correlation was observed between MCC and serum albumin (p = 0.06; r: -0.5459). Kidney biopsies of patients with nephrotic syndrome had higher MCC [2.12 (0.41-5.140) vs. 0.53 (0.0-3.94), p = 0.07]. Conclusion: Inflammatory cell infiltration and morphological differences between cell types in the inflammatory infiltrate are relevant factors in the assessment of the LN. Mast cell analysis and AI/CI assessment may be relevant prognostic indicators for pediatric patients with LN.

RESUMO Introdução: Mastócitos podem participar da inflamação e contribuir para fibrose na nefrite lúpica (NL). Objetivo: Correlacionar mastócitos em biópsias renais (BR) de pacientes pediátricos com NL com índices de atividade (IA) e cronicidade (IC), avaliando sua efetividade como fator prognóstico. Metodologia: Foram estudados 40 pacientes, entre 6 e 18 anos, diagnosticados com NL pelo Serviço de Nefropatologia da UFTM entre 1996 e 2015. Dados laboratoriais e epidemiológicos foram correlacionados com IA, IC e contagem de mastócitos (CM). Resultados: Encontramos correlação positiva e significativa entre contagem de mastócitos (CM) e IA (p = 0,003; r: 0,66) e entre CM e IC (p = 0,048; r: 0,48). Conforme a curva Roc, os mastócitos têm alta sensibilidade e especificidade na diferenciação de pacientes com IA menor ou maior do que 12. A creatinina sérica foi mais elevada na classe IV em relação à classe V [1,50 (0,40 - 20,90) versus 0,70 (0,62 - 0,90), p = 0,04]. Ureia sérica apresentou correlação positiva e significativa com CM (p = 0,002; r: 0,75). Observou-se tendência à correlação negativa entre CM e albumina sérica (p = 0,06; r: -0,5459). BR de pacientes com síndrome nefrótica apresentaram maior CM [2,12 (0,41 - 5,140) versus 0,53 (0,0 - 3,94), p = 0,07]. Conclusão: Não apenas o infiltrado inflamatório como também a diferenciação morfológica dos tipos celulares que o constituem são importantes para a avaliação da NL. Isso indica que a análise dos mastócitos, juntamente com a dos IA e IC, pode ajudar os nefrologistas a definirem o prognóstico de pacientes pediátricos.

Presence of langerhans cells, regulatory T cells (Treg) and mast cells in asymptomatic apical periodontitis

Abstract Asymptomatic Apical Periodontitis is essentially an inflammatory disease of microbial aetiology. Association and function of the cell components involved, or specific inductive factors and growth mediators associated with development, maintenance and resolution of the periapical lesions are still unknown. The objective of this study was to evaluate the concentration of Regulatory T cells (FoxP3+; Treg), Langerhans cells (CD1a+; LC) and mast cells in asymptomatic apical periodontitis. 73 cases were selected: 30 periapical granulomas, 29 radicular cysts and 14 residual cysts. All groups were submitted to morphological analysis for classification of inflammatory infiltrate and thickness of the epithelial lining as well as to immunohistochemical analysis for detection of LC and Treg cells. Toluidine blue staining was used for detecting mast cells. Analysis showed higher mean numbers of LC (8.2 cells/0.2mm2), and Treg cells in radicular cysts (5.910 cells/0.2mm2). As for mast cells, it was found that radicular cysts had a higher mean number of these cells compared to other periapical lesions (12.68 cells/0.2mm2). The association between thickness of the epithelial lining and inflammatory cells showed that the presence of hypertrophic epithelium in radicular cysts presented higher density of LC. The number of LC and Treg cells play an important role in the control of the inflammatory micro-environment in periapical granulomas and radicular cysts, respectively. The presence of mast cells in radicular cysts may be associated with progression of the lesion. Knowledge regarding the inflammatory cell profile is therefore essential for a better understanding of the pathogenesis of asymptomatic periapical periodontitis.

Although with intact mucosa at colonoscopy, chagasic megacolons have an overexpression of Gal-3 / Embora com mucosa íntegra à colonoscopia, megacólons chagásicos apresentam superexpressão de Gal-3

ABSTRACT Objective To evaluate the density of anti-galectin-3-immunostained cells, collagen percentage, mast cell density and presence of pathological processes in intestinal muscle biopsies of patients. Methods Thirty-five patients who underwent intestinal biopsy were selected from 1997 to 2015. Patients were divided into three groups: chagasic patients with mucosal lesion (n=13), chagasic patients with intact mucosa (n=12) and non-chagasic patients with no mucosal lesion (n=10). Histological processing of the biopsied fragments and immunohistochemistry for galectin-3 were performed. Additional sections were stained with hematoxylin and eosin to evaluate the general pathological processes, picrosirius for evaluation of collagen and toluidine blue to evaluate the mast cell density. Results Patients of mucosal lesion group had a significantly higher frequency of ganglionitis and myositis when compared to the chagasic patients with intact mucosa and non-chagasic group. The density of anti-galectin-3-immunostained cells was significantly higher in the chagasic patients with intact mucosa group when compared to the non-chagasic group. The group of chagasic patients with intact mucosa presented a higher percentage of collagen in relation to the patients with mucosal lesion and to the non-chagasic group, with a significant difference. There was no significant difference in mast cell density among the three groups. Conclusion The higher density of anti-galectin-3-immunostained cells in patients in the chagasic patients with intact mucosa group suggested the need for greater attention in clinical evaluation of these patients, since this protein is associated with neoplastic transformation and progression.

RESUMO Objetivo Avaliar a densidade de células imunomarcadas por anti-galectina-3, a percentagem de colágeno, a densidade de mastócitos e a presença de processos patológicos na musculatura intestinal de pacientes biopsiados. Métodos Foram selecionados 35 pacientes submetidos à biópsia de intestino entre 1997 a 2015. Os pacientes foram divididos em três grupos: chagásicos com lesão de mucosa (n=13), chagásicos com mucosa íntegra (n=12) e não chagásicos sem lesão de mucosa (n=10). Foram realizados processamento histológico dos fragmentos biopsiados e imunohistoquímica para galectina-3. Cortes adicionais foram corados por hematoxilina e eosina, para avaliar os processos patológicos gerais, pelo picrosírius, para avaliação do colágeno, e pelo azul de toluidina, para avaliar a densidade de mastócitos. Resultados Os pacientes do grupo chagásicos com lesão de mucosa apresentaram frequência significativamente maior de ganglionite e miosite quando comparados aos dos grupos chagásico com mucosa íntegra e não chagásicos. A densidade das células imunomarcadas por anti-galectina-3 foi significativamente maior no grupo chagásicos com mucosa íntegra quando comparada ao grupo não chagásico. O grupo de chagásicos com mucosa íntegra apresentou maior percentagem de colágeno em relação aos grupos chagásicos com mucosa lesada e ao grupo de não chagásicos, com diferença significativa. Não houve diferença significativa com relação à densidade de mastócitos entre os três grupos. Conclusão A maior densidade de células imunomarcadas por anti-galectina-3 nos pacientes do grupo chagásico com mucosa íntegra sugere a necessidade de maior atenção na avaliação clínica desses pacientes, uma vez que essa proteína está associada com transformação e progressão neoplásica.

Mast cells-derived MiR-223 destroys intestinal barrier function by inhibition of CLDN8 expression in intestinal epithelial cells

BACKGROUND: Mast cells (MCs) have been found to play a critical role during development of inflammatory bowel disease (IBD) that characterized by dysregulation of inflammation and impaired intestinal barrier function. However, the function of MCs in IBD remains to be fully elucidated. RESULTS: In our study, we used exosomes isolated from human mast cells-1 (HMCs-1) to culture with NCM460, HT-29 or CaCO2 of intestinal epithelial cells (lECs) to investigate the communication between MCs and lECs. We found that MCs-derived exosomes significantly increased intestinal epithelial permeability and destroyed intestinal barrier function, which is attributed to exosome-mediated functional miRNAs were transferred from HMCs-1 into lECs, leading to inhibit tight junction-related proteins expression, including tight junction proteins 1 (TJP1, ZO-1), Occludin (OCLN), Claudin 8 (CLDN8). Microarray and bioinformatic analysis have further revealed that a panel of miRNAs target different tight junction-related proteins. Interestingly, miR-223 is enriched in mast cell-derived exosome, which inhibit CLDN8 expression in IECs, while treatment with miR-223 inhibitor in HT-29 cells significantly reversed the inhibitory effect of HMCs-1-derived exosomes on CLDN 8 expression. Most importantly, enrichment of MCs accumulation in intestinal mucosa of patients with IBD compared with those healthy control. CONCLUSIONS: These results indicated that enrichment of exosomal miR-223 from HMCs-1 inhibited CLDN8 expression, leading to destroy intestinal barrier function. These finding provided a novel insight of MCs as a new target for therapeutic treatment of IBD.

Effect of pre-acupuncture intervention on serum IgE and cutaneous phosphorylated tyrosine-protein kinase expression in rats with urticaria / 针刺研究

OBJECTIVE: To observe the effect of acupuncture on serum IgE level, the degranulation of mast cells, the release of histamine and serotonin, and the expressions of phosphorylated tyrosine-protein kinase Lyn and Syk (p-Lyn, p-Syk) in skin tissue in rats with urticaria, as well as analyze the mechanism of acupuncture in the prevention and the treatment of urticaria. METHODS: SD male rats were randomly divided into normal control, model control, medication and acupuncture groups (n＝10 in each group). The anti-ovalbumin serum was used to establish urticaria model. Rats of the medication group received gastric lavage of Loratadine (0.1 mg/100 g). In the acupuncture group, bilateral "Xuehai" (SP10) and "Quchi" (LI11) were punctured perpendicularly, about 2 to 4 mm in depth, and the needles were retained for 30 min. The treatment was given consecutively for 14 days in the two treatment groups. H．E. staining was adopted to observe the morphological changes of skin tissue, ELISA to determine the total IgE level in serum, the toluidine blue staining to observe the degranulation of mast cells in local skin tissue and the immunohistochemistry to determine the expressions of histamine and serotonin as well as the the expressions of p-Lyn and p-Syk. RESULTS: Compared with the normal control group, the epidermis of the model control group was significantly thickened, the dermis was swollen, the inflammatory infiltration of small vessels was serious and the mast cells were swollen and deformed, with blurred edge and exfoliated granules. Additionally, in the model control group, the serum IgE level was significantly higher (P0.05). CONCLUSION: Acupuncture at LI11 and SP10 is applicable in the treatment of urticaria. This therapy inhibits the type Ⅰ hypersensitivity and the mast cell degranulation, which may be related to the regulation of p-Lyn and p-Syk protein expressions in the locus coeruleus skin tissue.

Effect of Medicated Serum Prepared with Chang'an Ⅰ Prescription on Sensitized Mast Cell Degranulation / 中国实验方剂学杂志

Objective:To observe the influence of Chang'an Ⅰ prescription drug-containing serum on IgE-mediated RBL-2H3 cell degranulation model， and explore the mechanism of Chang'an Ⅰ prescription in inhibiting RBL-2H3 activation degranulation and releasing inflammatory mediators with v-yes-1 Yanaguchi sarcoma viral related oncogene homolog （Lyn）/spleen tyrosine protein kinase （Syk）/mitogen-activated protein kinase （MAPK） signal pathway. Method:Preparation for Chang'an Ⅰ prescription serum. Animal group， SD male rats were randomly divided into Chang'an Ⅰ prescription serum high， medium， low dose， and blank control groups with 10 rats in each group. Dosage： 10 mL·kg-1 distilled water was given to blank control group， while Chang'an Ⅰ prescription serum high， medium and low dose groups were respectively given to the Chang'an Ⅰ prescription concentrated crude drug with concentration of 1.15，2.30，4.60 g·kg-1， respectively once a day for 7 days continuously and then blood was taken from aorta ventralis and centrifuged. Ketotifen as the positive control drug. Mast cells are counted with toluidine blue staining. Cellular release of β-aminohexose was detected by colorimetric method. Contents of MCT， TNF-α， MCP-1 and histamine were measured by enzyme-linked immunosorbent assay （ELISA） kits， Lyn/Syk/MAPK protein levels were detected by immunoblotting. Result:For cell activation and degranulation， compared with the blank control group， the model group had more cell degranulation （P<0.05）， compared with model group， the cell degranulation rate of each dose group of Chang'an Ⅰ prescription decreased （P<0.05）. The release rate of β-hexosamine in each dose group of Chang'an Ⅰ prescription decreased significantly （P<0.01）. For the release of active mediators， compared with the blank control group， the contents of histamine， MCT， TNF-α and MCP-1 all increased in the model group （P<0.01）， compared with the model group， the contents in each dose group of Chang'an Ⅰ prescription all decreased significantly （P<0.01）. Compared with the normal group， the phosphorylation levels of Lyn and Syk， extracellular regulatory protein kinase 1/2（ERK1/2）， c-Jun N-terminal kinase （JNK）， and mitogen-activated protein kinase p38 increased in the model group （P<0.05）. Compared with the model group， the Lyn， Syk and ERK1/2， JNK and p38 protein phosphorylation levels reduced in Chang'an Ⅰ prescription group （P<0.05）. Conclusion:Chang'an Ⅰ prescription drug-containing serum down-regulates the phosphorylation levels of proteins Lyn， Syk， and ERK1/2， JNK， and p38， inhibits RBL-2H3 cell activation and degranulation， reduces the release of cytokines and chemokines， such as histamine， MCT， TNF-α and MCP-1， it may be one of its mechanisms for treating IBS-D visceral hypersensitivity.