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Mast cells (MCs) are the principal effector cells of IgE-mediated allergy. IL-33 is released by resident skin cells as alarmin upon tissue damage or allergen contact. Owing to their pronounced receptor expression, MCs are important targets of IL-33 action, but consequences for skin MCs are ill-defined, especially upon chronic exposure to IL-33. Mimicking the inflammatory milieu of skin disorders, we found that persistent exposure to IL-33 (over a 5-week period) strengthened skin MC numbers through accelerated cell-cycle progression and restriction of apoptosis. Conversely, IL-33 attenuated degranulation and FcεRI expression, potentially as a feedback to chronic "alarmin" exposure. Interestingly, the negative impact on histamine release was counterbalanced by amplified histamine production. Considering the clinical significance of histamine and scarce information on its regulation, we explored the molecular underpinnings. IL-33 induced swift phosphorylation of p38 and JNK (but not of ERK1/2 or AKT), and stimulated histidine decarboxylase expression. Combining pharmacological inhibition and kinase elimination by Accell-facilitated RNA interference in skin MCs revealed a p38-dependent, but JNK-independent mechanism. Collectively, IL-33 exerts multifaceted effects on cutaneous MCs at a post-maturation stage. The IL-33-skin MC axis may contribute to and balance inflammation in chronic skin disorders., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Mast cells (MCs), unique cellular elements of the body, are commonly associated with IgE-mediated reactions and manifestations of Th2-type immunity. A key characteristic of the lineage is its heterogeneity, with subsets displaying significant variation depending on maturation stage, species, tissue, microenvironment and other. Heterogeneity also affects MC responses to extracellular cues. Indeed, IL-4, the signature cytokine of Th2-immunity, can affect MCs in opposing ways ranging from the induction of apoptosis to positive regulation of lineage characteristics. It is unknown, however, whether IL-4 alters the phenotype of terminally differentiated human cutaneous MCs. Using our well-established technique for homogeneous purification of human skin MCs, we now report that prolonged contact with IL-4 not only increases MC expansion, but also phenotypically and functionally re-shapes the cells. FcεRI cell surface expression, FcεRIα-specific mRNA and FcεRI-mediated histamine release are all augmented by IL-4, while histamine release elicited by the non-immunological stimulus, substance P, remains unaffected. IL-4's potential to mold MCs is broad and similarly detectable across donors. Intriguingly, IL-4 impacts granule-associated mediators, especially histamine whose synthesis is boosted in the presence of IL-4. To our knowledge, an increase in histamine production by IL-4 has not been described yet for any type of MCs, but may well contribute to its pro-allergic effect given the significance of this biogenic amine to allergic symptoms. Collectively, IL-4 alters human skin MCs after long-term exposure mimicking chronic disorders by strengthening MC numbers and intensifying processes associated with allergic inflammation.

Proteinase-activated receptor-2 (PAR2) belongs to a new G protein-coupled receptor subfamily activated by serine proteinases. PAR2 has been demonstrated to play a role during inflammation and immune response in different tissues including the skin. We examined whether PAR2 is functionally expressed by cutaneous human primary skin mast cells (HPMC) and the human mast cell line 1 (HMC-1). Reverse transcription-polymerase chain reaction and FACS analysis show expression of PAR2 both at the RNA and protein level. HPMCs and HMC-1 also express PAR1, PAR3, and PAR4. Ca-mobilization studies demonstrate functional PAR2 expressed by human skin mast cells, as shown by natural and synthetic PAR2 agonists. PAR2 agonists induced histamine release from HPMC indicating a role of PAR2 in regulating inflammatory and immune responses by skin mast cells. Double-immunofluorescence staining reveals colocalization of PAR2 with tryptase in the majority of human skin mast cells. In conclusion, trypsin and tryptase as well as specific agonists for PAR2 were able to induce Ca2+ mobilization in HPMCs, and agonists of PAR2 induce the release of histamine from these cells. Thus, PAR2 may be an important regulator of skin mast cell function during cutaneous inflammation and hypersensitivity.

Mast cells ( MCs) are unique constituents of the human body. While inter-individual differences may influence the ways by which MCs operate in their skin habitat, they have not been surveyed in a comprehensive manner so far. We therefore set out to quantify skin MC variability in a large cohort of subjects. Pathophysiologically relevant key features were quantified and correlated: transcripts of c-kit, Fc ε RI α, Fc ε RI β, Fc ε RI γ, histidine decarboxylase, tryptase, and chymase; surface expression of c-Kit, Fc ε RI α; activity of tryptase, and chymase; histamine content and release triggered by Fc ε RI and Ca2+ ionophore. While there was substantial variability among subjects, it strongly depended on the feature under study (coefficient of variation 33-386%). Surface expression of Fc ε RI was positively associated with Fc ε RI α mRNA content, histamine content with HDC mRNA, and chymase activity with chymase mRNA. Also, MC signature genes were co-regulated in distinct patterns. Intriguingly, histamine levels were positively linked to tryptase and chymase activity, whereas tryptase and chymase activity appeared to be uncorrelated. Fc ε RI triggered histamine release was highly variable and was unrelated to Fc ε RI expression but unexpectedly tightly correlated with histamine release elicited by Ca2+ ionophore. This most comprehensive and systematic work of its kind provides not only detailed insights into inter-individual variability in MCs, but also uncovers unexpected patterns of co-regulation among signature attributes of the lineage. Differences in MCs among humans may well underlie clinical responses in settings of allergic reactions and complex skin disorders alike. [ABSTRACT FROM AUTHOR]

The article offers information on the characteristics of mast cells in skin tissues and discusses the variability in them in different cohorts through an analysis. Topics discussed include the morphology of the cells, the granules in mast cells, and the differences in mast cells based on the location of the skin on the body.

Mast cells generate and release histamine during anaphylactic reactions, and there is pharmacological evidence that histamine regulates this process via specific receptors. Therefore, we examined human leukemic (HMC-1) and normal skin mast cells for the expression of all four currently known histamine receptors. Both cell types expressed H2 and H4 receptors at mRNA and protein levels, whereas H3 receptor specific mRNA and receptor protein was undetectable. Similarly, immunohistochemistry of cutaneous tissue showed an absence of H3 receptor in these cells. Despite transcription of mRNA, H1 receptor protein was only moderately expressed in HMC-1 cells and was virtually absent in skin mast cells. Furthermore, only H1, H2, and H4 receptors were detectable by Western blot analysis of HMC-1 cells. Radiolabeled histamine binding was strongly inhibited only by H2 (ranitidine)- and H3/H4 (FUB 108)-specific antagonists. Histamine-induced increase of cAMP was inhibited by the H2 receptor antagonist famotidine, whereas induction of IP3 was not observed, making signaling via the H1 receptor unlikely. These data show that human mast cells constitutively express primarily H2 and H4 receptors and that H2 receptors are functionally linked to cellular processes. They provide new insights into the mechanisms that govern auto- and paracrine histamine-induced mast cell functions. [ABSTRACT FROM AUTHOR]

As mast cells have been implicated in cutaneous repair processes, we have examined the ability of human mast cells to produce important epithelial and fibroblast growth factors or to stimulate the production of such factors in dermal fibroblasts. Isolated, highly purified human dermal mast cells and human leukemic mast cells were examined for mRNA and partly also for protein expression of these molecules as such or after preincubation with interleukin-4, stem cell factor, or with phorbol myristate acetate. In addition, mast cells were studied for their ability to induce fibroblast growth factor 2 and fibroblast growth factor 7 secretion from dermal fibroblasts. Both dermal and leukemic mast cells expressed fibroblast growth factor 2, fibroblast growth factor 7, and heparin-binding epidermal growth factor, but not hepatocyte growth factor at mRNA level, and dermal mast cells expressed fibroblast growth factor 10 in addition. At protein level, spontaneous fibroblast growth factor 2 secretion was noted that was markedly enhanced by phorbol myristate acetate, whereas no fibroblast growth factor 7 protein was detected under these conditions. Instead, human mast cell-1 supernatants induced enhanced fibroblast growth factor 7 secretion from dermal fibroblasts, with phorbol-myristate-acetate-stimulated supernatants being more effective. This effect could be reproduced with histamine and was H1-receptor mediated. Tryptase was ineffective but stimulated instead fibroblast growth factor 2 secretion from fibroblasts. These data demonstrate for the first time the ability of mast cells to express and/or secrete several growth factors of the fibroblast growth factor family as well as heparin-binding epidermal growth factor directly or indirectly via stimulation of fibroblasts, underlining the potentially pivotal role of these cells during human tissue repair and homeostasis. [ABSTRACT FROM AUTHOR]

Please cite this paper as: Mast cell lines HMC-1 and LAD2 in comparison with mature human skin mast cells – drastically reduced levels of tryptase and chymase in mast cell lines. Experimental Dermatology 2010; 19: 845–847. To circumvent the costly isolation procedure associated with tissue mast cells (MC), two human MC lines, i.e. HMC-1 and LAD2, are frequently employed, but their relation to mature MC is unknown. Here, we quantitatively assessed their expression of MC markers in direct comparison to skin MC (sMC). sMC expressed all lineage markers at highest and HMC-1 cells at lowest levels. LAD2 cells expressed comparable high-affinity IgE receptor α (FcεRIα) and FcεRIγ but less FcεRIβ than sMC and displayed slightly reduced, but robust FcεRI-mediated histamine release. Only minor differences were found for total histamine content and c-Kit expression. Huge, and to this level unexpected, differences were found for MC tryptase and chymase, with sMC >>> LAD2 > HMC-1. Taken together, HMC-1 cells represent very immature malignantly transformed MC, whereas LAD2 cells can be considered intermediately differentiated. Because of the minute levels of MC proteases, MC lines can serve as surrogates of tissue MC to a limited degree only. [ABSTRACT FROM AUTHOR]