Types Of Mast Cells Research Articles

Synthesis and Antiallergic Activity of Dicoumarin Derivatives.

Allergies are one of the diseases whose incidence rates have increased in recent years due to the greenhouse effect and extreme climate change. Therefore, the development of new antiallergic drugs has attracted the interest of researchers in chemistry and pharmacy fields. Dicoumarin is a coumarin derivative with various biological activities, but its antiallergic activity has not been evaluated. In this study, 14 different dicoumarin derivatives were synthesized by diethylamine-catalyzed condensation reactions of 4-hydroxycoumarin with 14 different aldehydes, and they were identified on the basis of their spectral data. The dicoumarin derivatives were subjected to studies on the degranulation of rat basophilic leukemia cells (RBL-2H3 cells) and mouse bone-marrow-derived mast cells (mBMMCs), and some of them showed good inhibitory effects on the degranulation of the two types of mast cells, demonstrating their good antiallergic activity. This study presents a new method of developing new antiallergic drugs.

Inhibition of mast cell degranulation by novel small molecule MRGPRX2 antagonists

BackgroundMas-related G-protein coupled receptor X2 (MRGPRX2) is a promiscuous receptor on mast cells that mediates IgE-independent degranulation and has been implicated in multiple mast cell-mediated disorders, including chronic urticaria, atopic dermatitis, and pain disorders. Although it is a promising therapeutic target, few potent, selective, small molecule antagonists have been identified, and functional effects of human MRGPRX2 inhibition have not been evaluated in vivo. ObjectiveWe identified and characterized novel, potent, and selective orally active small molecule MRGPRX2 antagonists for potential treatment of mast cell-mediated disease. MethodsAntagonists were identified using multiple functional assays in cell lines overexpressing human MRGPRX2, LAD2 mast cells, human peripheral stem cell-derived mast cells, and isolated skin mast cells. Skin mast cell degranulation was evaluated in Mrgprb2em(-/-) knockout (KO) and Mrgprb2em(MRGPRX2) transgenic human MRGPRX2 knock-in (KI) mice by assessment of agonist-induced skin vascular permeability. Ex vivo skin mast cell degranulation and associated histamine release was evaluated by microdialysis of human skin tissue samples. ResultsMRGPRX2 antagonists potently inhibited agonist-induced MRGPRX2 activation and mast cell degranulation in all mast cell types tested, in an IgE-independent manner. Orally administered MRGPRX2 antagonists also inhibited agonist-induced degranulation and resulting vascular permeability in MRGPRX2 KI mice. In addition, antagonist treatment dose dependently inhibited agonist-induced degranulation in ex vivo human skin. ConclusionMRGPRX2 small molecule antagonists potently inhibited agonist-induced mast cell degranulation in vitro and in vivo as well as ex vivo in human skin, supporting potential therapeutic utility as a novel treatment for multiple human diseases involving clinically relevant mast cell activation.

Ethanol Extract of Ampelopsis brevipedunculata Rhizomes Suppresses IgE-Mediated Mast Cell Activation and Anaphylaxis.

More than 20% of the world's population suffers from allergic diseases, including allergic asthma, rhinitis, and atopic dermatitis that severely reduce the patient's quality of life. The treatment of allergy has been developed, but there are still unmet needs. Ampelopsis brevipedunculata (Maxim.) Trautv. is a traditional medicinal herb with beneficial bioactivities, such as antioxidant, anti-hypertension, anti-viral, anti-mutagenic, and skin and liver (anti-hepatotoxic) protective actions. However, its anti-allergic effect has not been addressed. This study designed to investigate the pharmacological effect of an ethanol extract of A. brevipedunculata rhizomes (ABE) on mast cell and anaphylaxis models. For in vivo studies, we used ovalbumin-induced active systemic anaphylaxis (ASA) and immunoglobulin (Ig) E-mediated passive cutaneous anaphylaxis (PCA) models. In ASA model, oral administration of ABE (1, 10, and 100 mg/kg) attenuated the anaphylactic responses, such as hypothermia, serum histamine, and IgE productions. In PCA model, ABE also suppressed the plasma extravasation and swelling. The underlying mechanisms of action were identified in various mast cell types. In vitro, ABE (10, 30, and 60 µg/mL) inhibited the release of essential allergic mediators, such as histamine and β-hexosaminidase, in a concentration-dependent manner. ABE prevented the rapid increase in intracellular calcium levels induced by the DNP-HSA challenge. In addition, ABE downregulated the tumor necrosis factor-α and interleukin-4 by suppressing the activation of nuclear factor-κB. Collectively, this study is the first to identify the anti-allergic effect of ABE, suggesting that ABE is a promising candidate for treating allergic diseases.

A Single-Cell Transcriptome of Bovine Milk Somatic Cells.

The production of milk by dairy cows far exceeds the nutritional needs of the calf and is vital for the economical use of dairy cattle. High milk yield is a unique production trait that can be effectively enhanced through traditional selection methods. The process of lactation in cows serves as an excellent model for studying the biological aspects of lactation with the aim of exploring the mechanistic base of this complex trait at the cellular level. In this study, we analyzed the milk transcriptome at the single-cell level by conducting scRNA-seq analysis on milk samples from two Holstein Friesian cows at mid-lactation (75 and 93 days) using the 10× Chromium platform. Cells were pelleted and fat was removed from milk by centrifugation. The cell suspension from each cow was loaded on separate channels, resulting in the recovery of 9313 and 14,544 cells. Library samples were loaded onto two lanes of the NovaSeq 6000 (Illumina) instrument. After filtering at the cell and gene levels, a total of 7988 and 13,973 cells remained, respectively. We were able to reconstruct different cell types (milk-producing cells, progenitor cells, macrophages, monocytes, dendritic cells, T cells, B cells, mast cells, and neutrophils) in bovine milk. Our findings provide a valuable resource for identifying regulatory elements associated with various functions of the mammary gland such as lactation, tissue renewal, native immunity, protein and fat synthesis, and hormonal response.

Amyloid Beta Peptides Lead to Mast Cell Activation in a Novel 3D Hydrogel Model.

Alzheimer's disease (AD) is a prevalent neurodegenerative disease and the world's primary cause of dementia among the elderly population. The aggregation of toxic amyloid-beta (Aβ) is one of the main pathological hallmarks of the AD brain. Recently, neuroinflammation has been recognized as one of the major features of AD, which involves a network of interactions between immune cells. The mast cell (MC) is an innate immune cell type known to serve as a first responder to pathological changes and crosstalk with microglia and neurons. Although an increased number of mast cells were found near the sites of Aβ deposition, how mast cells are activated in AD is not clear. We developed a 3D culture system to culture MCs and investigated the activation of MCs by Aβ peptides. Because collagen I is the major component of extracellular matrix (ECM) in the brain, we encapsulated human LADR MCs in gels formed by collagen I. We found that 3D-cultured MCs survived and proliferated at the same level as MCs in suspension. Additionally, they can be induced to secrete inflammatory cytokines as well as MC proteases tryptase and chymase by typical MC activators interleukin 33 (IL-33) and IgE/anti-IgE. Culturing with peptides Aβ1-42, Aβ1-40, and Aβ25-35 caused MCs to secrete inflammatory mediators, with Aβ1-42 inducing the maximum level of activation. These data indicate that MCs respond to amyloid deposition to elicit inflammatory responses and demonstrate the validity of collagen gel as a model system to investigate MCs in a 3D environment to understand neuroinflammation in AD.

Mast cells in type 2 skin inflammation: Maintenance and function.

Mast cells (MCs) are immune cells residing in tissues and playing indispensable roles in maintaining homeostasis and inflammatory states. Skin lesions associated with atopic dermatitis (AD) and type 2 skin inflammation display an increment in MCs, which have both pro- and anti-inflammatory effects. The direct and indirect activations of skin MCs by environmental factors such as Staphylococcus aureus can instigate type 2 skin inflammation in AD with poorly understood mechanisms. Furthermore, both IgE-dependent and -independent degranulation of MCs contribute to pruritus in AD. Conversely, MCs suppress type 2 skin inflammation by promoting Treg expansion through IL-2 secretion in the spleen. Moreover, skin MCs can upregulate gene expression involved in skin barrier function, thus mitigating AD-like inflammation. These functional variances of MCs in AD could stem from differences in experimental systems, their localization, and origins. In this review, we will focus on how MCs are maintained in the skin under homeostatic and inflammatory conditions, and how they are involved in the pathogenesis of type 2 skin inflammation.

Oleanolic Acid Acetate Inhibits Mast Cell Activation in Ovalbumin-Induced Allergic Airway Inflammation.

Asthma is a complex, heterogeneous chronic inflammatory airway disease with multiple phenotypes. There has been a great progress in managing asthma, but there are still unmet needs for developing uncontrolled asthma treatments. The present study aimed to determine the effectiveness of oleanolic acid acetate (OAA) from Vigna angularis against allergic airway inflammation and the underlying mechanism of action with a focus on mast cells. To investigate the effect of OAA in allergic airway inflammation, we used the ovalbumin (OVA)-sensitized and challenged mice. To examine allergic airway inflammation associated with immune responses of mast cell activation in vitro, various types of mast cells were used. Systemic and cutaneous anaphylaxis models were used for mast cell-mediated hyper-responsiveness in vivo. OAA reduced OVA-induced airway inflammatory responses such as bronchospasm, increase of immune cell infiltration and serum immunoglobulin E and G1 levels. Especially, OAA decreased the mast cell infiltration, and β-hexosaminidase release as a mast cell activation marker in the bronchoalveolar lavage fluid. OAA inhibited mast cell degranulation in mast cell line (RBL-2H3) and primary cells (rat peritoneal mast cell and mouse bone marrow-derived mast cell). Mechanistically, OAA suppressed intracellular signaling pathways including the phosphorylation of phospholipase Cγ and nuclear factor-κB, resulting from the suppression of intracellular calcium influx and pro-inflammatory cytokine expression. Further, oral administration of OAA attenuated mast cell-mediated systemic and cutaneous anaphylaxis. Our study showed that OAA can inhibit mast cell-mediated allergic reaction. Consequently, the application of OAA to mast cells for the allergic airway inflammation facilitate a new direction of treating allergic asthma.

Mast cell distribution in porcine common bile duct with special reference to ghrelin

Due to the similar pattern of ghrelin localisation in human and porcine intestine, swine are described as a suitable animal model for experiments related to ghrelin and its receptors. In a previous study of ours, the presence of ghrelin immunoreactive endocrine cells in common bile duct (Ductus choledochus) (CBD) of domestic swine was established. However, data regarding the distribution of mast cell types in the CBD in this species are missing. The current study aimed to determine the distribution of tryptase-, ghrelin- and toluidine blue positive mast cells in the wall of extra- and intramural parts of porcine CBD. For this purpose, immunohistochemical staining for detection of tryptase was performed in order to identify ghrelin reactivity in mast cells. Additionally, toluidine blue staining was carried out to compare the number of metachromatic mast cells with that of tryptase positive cells. The comparative quantitative analysis showed that the number of tryptase immunorective cells was higher than that of metachromatic ones in the CBD wall. Ghrelin immunoreactive cells were the most numerous, therefore it can be assumed that not only mast cells but other cells also contain ghrelin. The three mast cells types were more abundant in the propria and subserosa of the intramural part than in the same layers of extramural CBD part. In the muscle layer, mast cell number was higher in the extramural than in the intramural part of the duct. In conclusion, taking into account the results obtained, an important role of mast cells play in controlling CBD motility and biliary gland function was suggested.

Mast cell density in domestic swine common hepatic duct

Detailed mast cell classification is reported in humans and rats, however such classification is not available in porcine common hepatic duct. It is interesting to find out whether mast cells in common hepatic duct are able to produce ghrelin, which is involved in a series of biological functions including regulation of food intake, body weight, gastrointestinal motility, hormone secretion, glucose release, cardiovascular functions, enzyme release, cell proliferation and reproduction in pigs. Because the determination of the cellular mechanisms responsible for normal and pathological biliary tract motility is difficult in humans, swine appears to be one of the most suitable animal model for physiological and morphological studies related to ghrelin peptide. The lack of information about the distribution of mast cells in the wall of common hepatic duct (Ductus hepaticus communis) in domestic swine motivated us to undertake this study in order to gain a better understanding of ghrelin peptide. The aim of this study was to define the localisation and number of tryptase-, ghrelin- and toluidine blue positive mast cells in the layers of porcine common hepatic duct. In this respect, the immunohistochemical staining for detection of tryptase and ghrelin expression was performed. The toluidine blue staining allowed the visualisation of metachromatic mast cells. The comparative study showed that tryptase positive cells were more numerous than metachromatic cells in all layers of the studied organ but the density of ghrelin positive cells was the highest. Tryptase immunohistochemistry allowed distinguishing ghrelin expressing mast cells from all ghrelin positive cells. The highest number of ghrelin positive cells indicated that not only mast cells but other cells also express ghrelin. The results allowed concluding that studied mast cell types had different density in the three main layers of common hepatic duct. The highest density of mast cells in the muscular layer and the possibility of mast cells to express ghrelin define its role in regulation of common hepatic duct motility and glandular secretion.

Antioxidant effect evaluation of drugs with different chemical structures by the degree of mast cell degranulation under photodynamic damage

Introduction. This model of skin acute inflammation caused by photodynamic damage (PHD), where reactive oxygen species (ROS) play a key role, enables the analysis of the microcirculation (MCC) dysfunction and degranulation of mast cells (MCs) at the site of exposure. The current study explored the IgE-independent mechanisms of MCs activation caused by PHD and the possibility of its pharmacological correction. Aim of the study – to evaluate the possibilities of using the model of acute inflammation induced by ROS during PHD to study the MCs contribution to the regulation of vascular permeability and to study angioprotective and MCC-improving drugs at the preclinical stage. Materials and methods. Male Wistar rats were injected with a photosensitizer, then anesthetized and laser irradiated 3 hours later, followed by one of the following drugs: hydrocortisone (HC), ethylmethylhydroxypyridine succinate (ES), or quinacrine (QC). Skin MCC was investigated by laser Doppler flowmetry. Calculation and morphometry of MCs was carried out on film preparations of loose connective tissue of the skin. Results. Immediately after PHD, the blood flow in the control group was 1.9 [1.4; 2.3] p. u., which is 55 % less than the initial values. Partial restoration of blood flow up to 3.7 [3.3; 4.0] p.u. was observed after one hour of observation (88 % of baseline, p<0.001). Despite the administration of HC and ES, the blood flow after PHD decreased by 8,5 and 32,5 %, respectively. After an hour, it was only 78 % of the baseline. Intravenous administration of QC immediately after irradiation, lead to decrease of the blood flow only 28 %, and after an hour the blood flow was completely restored. The degree of MCs degranulation after the intravenous administration of HC and QC is almost equal and characterized by a decrease in the number of MCs with complete (anaphylactic) degranulation to 27.5 [21.6; 29.4] and 26.4 [22.5; 32.5] %, respectively, versus 46.9 [47.7; 52] % in the control group (p<0,05); however, after the administration of ES, the results are comparable with the intact control. Non-parametric correlation analysis did not reveale statistically significant difference between blood flow one hour after photodynamic exposure and morphometric types of MCs in groups with various drugs. Conclusion. Differences between the drug effects on the skin blood flow and the IgE-independent MCs activation is confirmed by the absence of a correlation between these parameters. QC, in comparison with ES and HC, is more effective in relation to dysfunction of the skin MCC. Under these conditions, the combined use of anti-inflammatory and antioxidant drugs seems promising.

Evidence underscoring immunological and clinical pathological changes associated with Sarcoptes scabiei infection: synthesis and meta-analysis

BackgroundSarcoptes scabiei is one of the most impactful mammalian parasites. There has been much research on immunological and clinical pathological changes associated with S. scabiei parasitism across a range of host species. This rich body of literature is complex, and we seek to bring that complexity together in this study. We first (1) synthesise narrative reviews of immunopathological relationships to S. scabiei infection to construct overarching hypotheses; then (2) undertake a systematic meta-analysis of primary literature on immunological and clinical pathological changes; and lastly (3) contrast our findings from the meta-analysis to our synthesis from narrative reviews.MethodsWe synthesised 55 narrative reviews into two overarching hypotheses representing type I and type IV immune responses to S. scabiei infection. We then systematically extracted all literature reporting immunological variables, acute phase proteins, oxidant/antioxidant status, and erythrocytic, hepatological and nephrological changes, calculating 565 effect sizes between controls and sarcoptic mange affected groupings, refining (simplifying) hypotheses from narrative reviews.ResultsImmunological and clinical pathological parameters were most often studied in dogs (n = 12) and humans (n = 14). Combining immunological and clinical pathological information across mammalian species (n = 19) helped yield general insights into observed disease responses. This is evidenced by interspecific consensus in 27 immunological and clinical pathology variables (6/26 type I hypersensitivity, 3/20 type IV hypersensitivity, 6/10 oxidant/antioxidant status, 3/6 acute phase protein, 4/7 erythrocytic, and 5/10 hepatological/nephrological).ConclusionsElevated IgE, eosinophils and mast cells in type I hypersensitivity response corresponded to what was described in narrative reviews. Results from type IV hypersensitivity response suggested typical antibody response, however cell-mediated response was less evident. Some consensus of acute phase protein response and shifted oxidant/antioxidant balance and slight evidence of anemia. We highlight the need for mange/scabies studies to more routinely compare immunological and clinical pathological changes against controls, and include collection of a more standardised suite of variables among studies.

Coactivity of Mast Cells and Stem Cells on Angiogenesis and Antioxidants' Potentials at Inflammation, Proliferation, and Tissue Remodeling Phases of Wound.

Background Reactive oxygen species cause serious damage to the physiological function of tissues. Determination of total antioxidant capacity of skin tissue is one of the determinants of damaged tissue function. Mast cells (MCs) are one of the groups of cells that are invited to the site of injury. The healing process begins with the rapid release of various types of MCs' intermediate factors at the site of injury. Bone marrow mesenchymal stem cell (BMMSC) production and secretion have been shown to regenerate the skin. The aim of this research was to evaluate the wound-healing and antioxidant effects of BMMSCs per MCs. Methods Fifty-four albino Wistar male rats were divided into three groups: (1) nonsurgery, (2) surgery, and (3) surgery + BMMSCs. Groups 2 and 3 were operated with a 3 × 8 cm flap and in group 3, cell injections (7 × 10 9 cell injection at the time of surgery) were performed. After days 4, 7, and 15, percentage of the surviving tissue, histological characteristics, superoxide dismutase (SOD) activity, and amount of malondialdehyde (MDA) were measured in the groups. For results, Graph Pad Prism 8 software was used, and data were analyzed and compared by analysis of variance and Tukey test. Results BMMSCs' application decreased the amount of MDA, increased SOD activity and survival rate of the flaps, and improved the histological characteristics. Conclusion This study revealed the protective effects BMMSCs alongside MCs against oxidative stress on the survival of the flaps. However, for clinical use, more research is needed to determine its benefits.

Mast Cells Drive Systemic Inflammation and Compromised Bone Repair After Trauma.

There is evidence that mast cells contribute to inflammation induced by hemorrhagic shock, severe tissue injury or sepsis. Mast cells are highly responsive to alarm signals generated after trauma, and release many inflammatory mediators including interleukin-6, a key mediator of posttraumatic inflammation. An overwhelming posttraumatic inflammation causes compromised bone healing; however, the underlying cellular and molecular mechanisms are poorly understood. Recently, we found that mast cells trigger local and systemic inflammation after isolated fracture leading to uneventful bone repair. Here, we investigated whether mast cells critically contribute to trauma-induced compromised bone healing. Male Mcpt5-Cre+ R-DTA mice, which lack connective tissue type mast cells, and their mast cell-competent Cre− littermates underwent a femur fracture with/without thoracic trauma. Posttraumatic systemic and local inflammation and bone repair were assessed 3 h and 21 d post injury. Both, the systemic and pulmonary inflammation was significantly increased in mast cell-competent mice upon combined trauma compared to isolated fracture. In mast cell-deficient mice, the increase of inflammatory mediators in the circulation induced by the severe trauma was abolished. In the bronchoalveolar lavage fluid, the trauma-induced increase of inflammatory cytokines was not reduced, but the neutrophil invasion into the lungs was significantly diminished in the absence of mast cells. Locally in the fracture hematoma, mast cell-competent mice displayed reduced inflammatory mediator concentrations after combined trauma compared to isolated fracture, which was abolished in mast cell-deficient mice. Notably, while combined trauma resulted in compromised bone repair in mast cell-competent mice, indicated by significantly reduced bone and increased cartilage fracture callus contents, this was abolished in Mcpt5-Cre+ R-DTA mice. Therefore, mast cells contribute to trauma-induced compromised bone repair and could be a potential target for new treatment options to improve fracture healing in multiply injured patients.

In Obese Patients With Type 2 Diabetes, Mast Cells in Omental Adipose Tissue Decrease the Surface Expression of CD45, CD117, CD203c, and FcϵRI.

The paradigm of mast cells in type 2 diabetes is changing. Although they were first considered deleterious inflammatory cells, now they seem to be important players driving adipose tissue homeostasis. Here we have employed a flow cytometry-based approach for measuring the surface expression of 4 proteins (CD45, CD117, CD203c, and FcϵRI) on mast cells of omental (o-WAT) and subcutaneous white adipose tissue (s-WAT) in a cohort of 96 patients with morbid obesity. The cohort was split into three groups: non-T2D, pre-T2D, and T2D. Noteworthy, patients with T2D have a mild condition (HbA1c <7%). In o-WAT, mast cells of patients with T2D have a decrease in the surface expression of CD45 (p=0.0013), CD117 (p=0.0066), CD203c (p=0.0025), and FcϵRI (p=0.043). Besides, in s-WAT, the decrease was seen only in CD117 (p=0.046). These results indicate that T2D affects more to mast cells in o-WAT than in s-WAT. The decrease in these four proteins has serious effects on mast cell function. CD117 is critical for mast cell survival, while CD45 and FcϵRI are important for mast cell activation. Additionally, CD203c is only present on the cell surface after granule release. Taking together these observations, we suggest that mast cells in o-WAT of patients with T2D have a decreased survival, activation capacity, and secretory function.

Fetal Tissue-Derived Mast Cells (MC) as Experimental Surrogate for In Vivo Connective Tissue MC.

Bone-marrow-derived mast cells are matured from bone marrow cells in medium containing 20% fetal calf serum (FCS), interleukin (IL)-3 and stem-cell factor (SCF) and are used as in vitro models to study mast cells (MC) and their role in health and disease. In vivo, however, BM-derived hematopoietic stem cells account for only a fraction of MC; the majority of MC in vivo are and remain tissue resident. In this study we established a side-by-side culture with BMMC, fetal skin MC (FSMC) or fetal liver MC (FLMC) for comparative studies to identify the best surrogates for mature connective tissue MC (CTMC). All three MC types showed comparable morphology by histology and MC phenotype by flow cytometry. Heterogeneity was detected in the transcriptome with the most differentially expressed genes in FSMC compared to BMMC being Hdc and Tpsb2. Expression of ST2 was highly expressed in BMMC and FSMC and reduced in FLMC, diminishing their secretion of type 2 cytokines. Higher granule content, stronger response to FcεRI activation and significantly higher release of histamine from FSMC compared to FLMC and BMMC indicated differences in MC development in vitro dependent on the tissue of origin. Thus, tissues of origin imprint MC precursor cells to acquire distinct phenotypes and signatures despite identical culture conditions. Fetal-derived MC resemble mature CTMC, with FSMC being the most developed.

Mastocytosis patients' cognitive dysfunctions correlate with the presence of spindle-shaped mast cells in bone marrow.

To the Editor, Mastocytosis is a hematological neoplasm with a broad spectrum of disease caused by mast cell (MC) infiltration (i.e., impairment of function of bone marrow, liver) and degranulation (i.e., anaphylaxis).1 Recent data indicate psychological factors and neurological factors.2 Patients with mastocytosis have a significantly lower quality of life and experience increased anxiety, depression, and other symptoms such as headache or fatigue.3, 4 These might be accompanied by cognitive impairment and brain fog declared by 86% of US patients, though its relation with mastocytosis and its progression hasn't been established to date.5 Some of these symptoms might result from abnormal MCs in central nervous system.6 In murine model, Esposito et al. showed that stress through corticotropin stimulates brain MCs to disrupt permeability of brain–blood barrier. This effect was absent in MC-depleted rodents and after cromolyn administration. The aim of the study was to investigate cognitive dysfunctions in patients with mastocytosis and analyze its connection with disease characteristics in patients. The group of 79 Polish patients suffering from mastocytosis was enrolled (59 women and 20 men; age 20–75) after signing written informed consent. Fifty-four of them suffered from indolent systemic mastocytosis, 2 had diffuse cutaneous mastocytosis, 20 had maculopapulous cutaneous mastocytosis, 2 had smoldering systemic mastocytosis, and there was a single patient with bone marrow mastocytosis. Serum tryptase ranged from 3 to 190 ng/ml with mean value 47.6 ng/ml. The education level of the studied population was high. Forty patients reported high education, 26 middle school, 10 vocational education, and 3 elementary. All of the patients were evaluated with Mini–Mental State Examination (MMSE) that assesses cognitive functions in 0–30 point scale.7 The cut-off point that suggests dementia is below 24 for educated populations, including Poland. All patients had bone marrow biopsy performed. Mastocytosis treatment was evaluated with WHO Performance Status; the patients were also asked about particular typical MC activation symptoms.8 Statistical analysis was conducted with Statistica software using Spearman correlation test. Cut-off point for the p-value was 0.05. The mean MMSE score was 27.8 (median 29). Six out of 79 patients (7.6%) scored below 24 points (see Figure 1). Other 16 patients (20%) scored 24–26 points, which suggests minimal cognitive impairment (MCI).7 According to performance status, 23 patients had no symptoms, 55 scored 1, and one patient scored 2. There was a positive correlation with education level (R = 0.45) and a negative one with age (R = −0.34), as expected. MMSE correlated negatively with the presence of 25% atypical spindle-shaped MCs in bone marrow (minor systemic mastocytosis criterion) with (R = −0.5) and bone pain (R = −0.45). No correlation was found between MMSE and serum tryptase level, presence of KIT bone marrow mutation, CD2/CD25 expression, skin involvement or performance status. A weak correlation was observed for the presence of CD30 (R = −0.1) and symptoms: pruritus (R = −0.13), edema (R = −0.23). There were no differences in results between patients with cutaneous and indolent systemic mastocytosis subgroups. Number of cases with results of Mini-Mental State Examination. Red color shows potential dementia; yellow color shows minimal cognitive impairment; and blue color shows normal results The results of our study are in contrast to previously done surveys by Jennings et al.5 where 86% of the sample declared cognitive dysfunctions. However, our study suggests that such high cognitive impairment prevalence could have been over-diagnosed. Using well-known, validated MMSE method, we observed that only 7.6% of the sample experienced significant cognitive impairment while total 27% presented slight cognitive abnormalities. Such high self-reported symptoms may be due to multiple emotional and personal problems, or affective disorders, that patients experience because of MC activation. Those might be misinterpreted by patients and labeled as cognitive dysfunctions. The percentage of atypical MCs and bone pain that correlated to MMSE is associated with systemic mastocytosis and usually increases as the disease progresses to an aggressive form. Interestingly, there was no relation of MMSE to serum tryptase concentration, skin involvement, and general symptoms assessed by performance status. It is possible that cognitive dysfunctions might be linked to mastocytosis aggressiveness rather than to MC activation. In such case, according to Theoharides et al., serum IL-6 might be a promising predictor of cognitive dysfunctions as it is related to MC-related osteoporosis and bone pain.9 However, further studies are required in this area. education, mast cells, quality of life bildung, lebensqualita, mastozytose, mastzelle Publication costs were covered by Medical University of Gdansk statutory funds ST 02-141/07/231. The authors declare no conflict of interests. Medical University of Gdansk, Grant/Award Number: ST 02-141/07/231

DISTRIBUTION AND QUANTITATIVE CHANGES OF MAST CELLS IN GUINEA PIGS LUNG IN OVALBUMIN-INDUCED ALLERGIC INFLAMMATION

Background. One of the most important cells in local immunity in lung are mast cells. They are involved in both innate and adaptive immune responses to inhaled allergens. The question of distribution of these both cell types in guinea pig lung in case of experimental allergic inflammation in most aspects remains open. Objective. The aim of this research is to study the distribution and quantitative changes of mast cells in lung of guinea pigs in ovalbumin-induced allergic inflammation. Methods. the lungs of 48 male guinea pigs have been studied using histological, morphometric and statistical methods in cases of experimental ovalbumin-induced allergic inflammation. The total number of mucosa related must cells and perivascular mast cells in guinea pig lungs were counted. Results. It has been established that mucosa related mast cells are normally more abundant in guinea pigs lung than perivascular ones. Maximum increase in a number of mucosa related mast cells was revealed in the early period of allergic inflammation, as evidenced by maximum increase coefficient of 1.4 in the 1st experimental group, compare to the control (P*/**&lt;0.05). However, maximum increase in number of perivascular mast cells in 5 times was found during the late period of allergic inflammation in the 4th experimental group (P*/**&lt;0.05). Conclusion. Experimental sensitization and challenge with ovalbumin leads to statistically significant increase in average number of both types of mast cells but predominantly the latter ones. It has been proved that cells dynamics is multidirectional.

Lactic Acid Bacteria Fermented Cordyceps militaris (GRC-SC11) Suppresses IgE Mediated Mast Cell Activation and Type I Hypersensitive Allergic Murine Model.

Cordyceps militaris (C. militaris) has various biomedical applications in traditional oriental medicine for different diseases including inflammatory and immune-dysregulated diseases. It is a reservoir of nutritional components such as cordycepin, polysaccharides, and antioxidants. To improve its bioactivity, we fermented C. militaris with a Pediococcus pentosaceus strain isolated from a salted small octopus (SC11). The current study aimed to evaluate whether P. pentosaceus (SC11) fermentation could enhance the anti-allergic potential of C. militaris cultured on germinated Rhynchosia nulubilis (GRC) against a type I hypersensitive reaction in in vitro and in vivo studies. Total antioxidant capacity and cordycepin content were significantly increased in GRC after SC11 fermentation. GRC-SC11 showed significantly enhanced anti-allergic responses by inhibiting immunoglobulin E (IgE)/antigen-induced degranulation in RBL-2H3 cells, compared to GRC. The results demonstrated the significant inhibition of phosphorylated spleen tyrosine kinase (Syk)/ p38/GRB2-associated binding protein 2 (Gab2)/c-jun in IgE/Ag-triggered RBL-2H3 cells. Furthermore, suppressed mRNA levels of interleukin-4 (IL-4) and tumor necrosis factor-α (TNF-α) in IgE/Ag-activated RBL-2H3 cells were observed. GRC-SC11 significantly ameliorated IgE-induced allergic reactions by suppressing the ear swelling, vascular permeability, and inflammatory cell infiltration in passive cutaneous anaphylaxis (PCA) BALB/c mice. In conclusion, GRC fermented with P.pentosaceus exerted enhanced anti-allergic effects, and increased the cordycepin content and antioxidants potential compared to GRC. It can be used as bio-functional food in the prevention and management of type I allergic diseases.

IgE cross-linking induces activation of human and mouse mast cell progenitors

The concept of innate and adaptive effector cells that are repleted by maturing inert progenitor cell populations is changing. Mast cells develop from rare mast cell progenitors populating peripheral tissues at homeostatic conditions, or as a result of induced recruitment during inflammatory conditions. Because FcεRI-expressing mast cell progenitors are the dominating mast cell type during acute allergic lung inflammation invivo, we hypothesized that they are activated by IgE cross-linking. Mouse peritoneal and human peripheral blood cells were sensitized and stimulated with antigen, or stimulated with anti-IgE, and the mast cell progenitor population analyzed for signs of activation by flow cytometry. Isolated peritoneal mast cell progenitors were studied before and after anti-IgE stimulation at single-cell level by time-lapse fluorescence microscopy. Lung mast cell progenitors were analyzed for their ability to produce IL-13 by intracellular flow cytometry in a mouse model of ovalbumin-induced allergic airway inflammation. Sensitized mouse peritoneal mast cell progenitors demonstrate increased levels of phosphorylation of tyrosines on intracellular proteins (total tyrosine phosphorylation), and spleen tyrosine kinase (Syk) phosphorylation after antigen exposure. Anti-IgE induced cell surface-associated lysomal-associated membrane protein-1 (LAMP-1) in naive mast cell progenitors, and prompted loss of fluorescence signal and altered morphology of isolated cells loaded with lysotracker. In human mast cell progenitors, anti-IgE increased total tyrosine phosphorylation, cell surface-associated LAMP-1, and CD63. Lung mast cell progenitors from mice with ovalbumin-induced allergic airway inflammation produce IL-13. Mast cell progenitors become activated by IgE cross-linking and may contribute to the pathology associated with acute allergic airway inflammation.

Mast Cells in Human Cutaneous Neurofibromas: Density, Subtypes, and Association with Clinical Features in Neurofibromatosis 1

Background: Cutaneous neurofibromas (cNFs) are hallmarks of neurofibromatosis 1 (NF1) and cause the main disease burden in adults with NF1. Mast cells are a known component of cNFs. However, no comprehensive characterization of mast cells in cNFs is available, and their contributions to cNF growth and symptoms such as itch are not known. Methods: We collected 60 cNFs from ten individuals with NF1, studied their mast cell proteinase content, and compared the mast cell numbers to selected clinical features of the tumors and patients. The tumors were immunolabeled for the mast cell markers CD117, tryptase, and chymase, and the percentage of immunopositive cells was determined using computer-assisted methods. Results: The median proportions of positive cells were 5.5% (range 0.1–14.4) for CD117, 4.0% (1.2–7.0) for tryptase, and 5.0% (1.1–15.9) for chymase. The median densities of cells immunopositive for CD117, tryptase, and chymase were 280, 243, and 250 cells/mm2, respectively. Small tumors, growing tumors, and tumors from patients below the median age of 33 years displayed a high proportion of mast cells. Cells expressing both tryptase and chymase were the predominant mast cell type in cNFs, followed by cells expressing chymase only. Conclusion: The results highlight the abundance of mast cells in cNFs and that their number and subtypes clearly differ from those previously reported in unaffected skin.