Mite allergen is a danger signal for the skin via activation of inflammasome in keratinocytes
ABSTRACT Atopic dermatitis (AD) is a chronic inflammatory skin disorder caused by multiple factors. Among them, house dust mite (HDM) allergens are important in the development of AD. In airway allergy, HDM allergens activate innate immunity. However, information regarding the activation of innate immunity by HDM allergens in the skin is limited.
The inflammasome is a key regulator of pathogen recognition and inflammation. We investigated whether HDM allergens activate the inflammasome in epidermal keratinocytes.
Keratinocytes were stimulated with Dermatophagoides pteronyssinus, and the activation of caspase-1 and secretion of IL-1β and IL-18 were examined. Formation of the inflammasome was studied by analyzing the subcellular distributions of inflammasome proteins. The importance of specific inflammasome proteins was studied by knocking down their expression through transfection of keratinocytes with lentiviral particles carrying short hairpin RNAs (shRNAs).
D pteronyssinus activated caspase-1 and induced caspase-1-dependent release of IL-1β and IL-18 from keratinocytes. Moreover, D pteronyssinus stimulated assembly of the inflammasome by recruiting apoptosis-associated specklike protein containing a caspase-recruitment domain (ASC), caspase-1, and nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin-domain containing 3 (NLRP3) to the perinuclear region. Finally, infection with lentiviral particles carrying ASC, caspase-1, or NLRP3 shRNAs suppressed the release of IL-1β and IL-18 from the keratinocytes. Activation of the NLRP3 inflammasome by D pteronyssinus was dependent on cysteine protease activity.
House dust mite allergens are danger signals for the skin. In addition, HDM-induced activation of the NLRP3 inflammasome may play a pivotal role in the pathogenesis of AD.
- SourceAvailable from: Masamoto Murakami
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- "Primary human keratinocytes were isolated from surgically discarded neonatal skin samples, and cultured in MCDB153 medium, supplemented with insulin (5 μg ml−1), hydrocortisone (5×10−7 M), ethanolamine (0.1 mM), phosphoethanolamine (0.1 mM), bovine hypothalamic extract (50 μg ml−1) and Ca2+ (0.03 mM), as described previously . Cells that had been passaged four times were used in the experiments, and subconfluent keratinocyte cultures were subjected to stimulation. "
ABSTRACT: Eccrine sweat is secreted onto the skin's surface and is not harmful to normal skin, but can exacerbate eczematous lesions in atopic dermatitis. Although eccrine sweat contains a number of minerals, proteins, and proteolytic enzymes, how it causes skin inflammation is not clear. We hypothesized that it stimulates keratinocytes directly, as a danger signal. Eccrine sweat was collected from the arms of healthy volunteers after exercise, and levels of proinflammatory cytokines in the sweat were quantified by ELISA. We detected the presence of IL-1α, IL-1β, and high levels of IL-31 in sweat samples. To investigate whether sweat activates keratinocytes, normal human keratinocytes were stimulated with concentrated sweat. Western blot analysis demonstrated the activation of NF-κB, ERK, and JNK signaling in sweat-stimulated keratinocytes. Real-time PCR using total RNA and ELISA analysis of supernatants showed the upregulation of IL-8 and IL-1β by sweat. Furthermore, pretreatment with IL-1R antagonist blocked sweat-stimulated cytokine production and signal activation, indicating that bioactive IL-1 is a major factor in the activation of keratinocytes by sweat. Moreover, IL-31 seems to be another sweat stimulator that activates keratinocytes to produce inflammatory cytokine, CCL2. Sweat is secreted onto the skin's surface and does not come into contact with keratinocytes in normal skin. However, in skin with a defective cutaneous barrier, such as atopic dermatitis-affected skin, sweat cytokines can directly act on epidermal keratinocytes, resulting in their activation. In conclusion, eccrine sweat contains proinflammatory cytokines, IL-1 and IL-31, and activates epidermal keratinocytes as a danger signal.PLoS ONE 07/2013; 8(7):e67666. DOI:10.1371/journal.pone.0067666 · 3.23 Impact Factor
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- "Wild-type and NLRP3-deficient mice developed comparable allergic airway inflammation induced by HDM extracts suggesting that NLRP3 does not contribute to the HDM allergic response [79, 80]. However, the activation of human keratinocytes with HDM extracts triggered caspase-1 activation and promoted the caspase-dependent release of IL-1β and IL-18 suggesting the importance of the inflammasome engagement in the pathogenesis of AD . The assembly of NLRP3 inflammasome through the HDM cysteine protease Der p 1 was clearly evidenced in those cells. "
ABSTRACT: Sensitizations to house dust mites (HDM) trigger strong exacerbated allergen-induced inflammation of the skin and airways mucosa from atopic subjects resulting in atopic dermatitis as well as allergic rhinitis and asthma. Initially, the Th2-biased HDM allergic response was considered to be mediated only by allergen B- and T-cell epitopes to promote allergen-specific IgE production as well as IL-4, IL-5, and IL-13 to recruit inflammatory cells. But this general molecular model of HDM allergenicity must be revisited as a growing literature suggests that stimulations of innate immune activation pathways by HDM allergens offer new answers to the following question: what makes an HDM allergen an allergen? Indeed, HDM is a carrier not only for allergenic proteins but also microbial adjuvant compounds, both of which are able to stimulate innate signaling pathways leading to allergy. This paper will describe the multiple ways used by HDM allergens together with microbial compounds to control the initiation of the allergic response through engagement of innate immunity.02/2013; 2013(4):735031. DOI:10.1155/2013/735031
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- "In particular, the disruption of the skin barrier by other stimulants such as volatile organic compounds, can enhance the adverse effects of HDM on provocation testing (22), suggesting that barrier dysfunction of AD skin lesion might lead to a direct pathophysiological role of HDM in AD patients. HDM allergens can be danger signals in the skin through nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin-domain containing 3 (NLRP 3), which plays a central role in both innate immunity and inflammation (23). The results obtained in our study along with previous reports indicate that the proteolytic activities of HDM allergens might play a role as a nonspecific irritant in AD patients. "
ABSTRACT: We attempted to investigate the correlation between the severity of atopic dermatitis (AD) in children and the indoor level of house dust mite (HDM) allergens. Ninety-five patients (31.1 ± 19.5 months of age) with AD were enrolled in this study, and serum specific IgE against Dermatophagoides pteronyssinus and D. farinae was measured. The severity of AD was assessed using the visual analogue scale on the same day of house dust collection. Living rooms and mattresses where the child usually slept were vacuumed for 2 minutes and concentrations of Der f 1 were measured by enzyme-linked immunosorbent assay. The skin symptoms were more severe in patients with Der f 1 concentrations in living room > 2 µg/g dust than ≤ 2 µg/g dust (P = 0.018). This difference was noted in AD patients without sensitization to HDM (P = 0.004), but not in patients with sensitization. There was no difference in symptom severity according to Der f 1 concentrations in mattresses (P = 0.062). The severity of skin symptoms is associated with indoor concentrations of HDM in children with AD, and it is likely to act as nonspecific irritants as well as allergens in AD skin lesions.Journal of Korean medical science 01/2013; 28(1):74-9. DOI:10.3346/jkms.2013.28.1.74 · 1.27 Impact Factor