Proteases in agricultural dust induce lung inflammation through PAR-1 and PAR-2 activation

Romberger, Debra J.; Heires, Art J.; Nordgren, Tara M.; Souder, Chelsea; West, William W.; Liu, Xiangde; Poole, Jill; Toews, Myron L.; Wyatt, Todd A.

doi:10.1152/ajplung.00025.2015

Cited by 34 publications

(40 citation statements)

References 45 publications

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“…Notwithstanding, overall, the disulphide-maintained conformation of all of the enzymes due to the 6 conserved cysteines would be expected to expose both common and distinct antigenic sites. Given the demonstrated reactivity of Per a 10-sensitized human subjects in terms of their intradermal response, IgE immunoreactivity and Per a 10-stimulated histamine release by peripheral blood leukocytes [8, 9], it is possible that there may be cross-sensitization to enzymes from all cockroach species and also possibly to mammalian and fish trypsins to which agricultural and fish plant workers are exposed [22, 23]. Whether or not the insect serine proteinases represent significant allergens themselves, the ones we have studied so far all appear to be able to signal via PAR2.…”

Section: Discussionmentioning

confidence: 99%

“…Given the demonstrated reactivity of Per a 10-sensitized human subjects in terms of their intradermal response, IgE immunoreactivity and Per a 10-stimulated histamine release by peripheral blood leucocytes, 8,9 it is possible that there may be cross-sensitization to enzymes from all cockroach species and also possibly to mammalian and fish trypsins to which agricultural and fish plant workers are exposed. 23,24 Whether or not the insect serine proteinases represent significant allergens themselves, the ones we have studied so far all appear to be able to signal via PAR2. Thus, all of the enzymes E1, E2 and E3 in isolation would be able to synergize with other allergens in the cockroach extract to trigger the allergic process.…”

Section: Erk 1/2 Via Par2mentioning

confidence: 97%

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Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Polley

Mihara

Ramachandran

et al. 2017

Clin Experimental Allergy

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SUMMARY BACKGROUND Allergy to the German cockroach (Blattella germanica) is a significant asthma risk factor for inner-city communities. Cockroach, like other allergens, contains trypsin-like enzyme activity that contributes to allergenicity and airway inflammation by activating proteinase-activated receptors (PARs). To date, the enzymes responsible for the proteolytic activity of German cockroach allergen have not been characterized. OBJECTIVES We aimed to identify, isolate and characterize the trypsin-like proteinases in German cockroach allergen extracts used for clinical skin tests. For each enzyme, we sought to determine (1) its substrate and inhibitor enzyme kinetics (Km and IC50); (2) its amino acid sequence and (3) its ability to activate calcium signaling and/or ERK1/2 phosphorylation via PAR2. METHODS Using a trypsin-specific activity-based probe, we detected three distinct enzymes that were isolated using ion-exchange chromatography. Each enzyme was sequenced by mass spectometery (deconvoluted with an expressed sequence tag library), evaluated kinetically for its substrate/inhibitor profile and assessed for its ability to activate PAR2 signaling. FINDINGS Each of the three serine proteinase-activity-based probe-labelled enzymes isolated were biochemically distinct, with different enzyme kinetic profiles and primary amino acid sequences. The three enzymes showed a 57 to 71% sequence identity with a proteinase previously cloned from the American cockroach (Per a 10). Each enzyme was found to activate both Ca++ and MAPK signaling via PAR2. CONCLUSIONS AND RELEVANCE We have identified three different serine proteinases from the German cockroach that may, via PAR2 activation, play different roles for allergen-sensitization in vivo and may represent attractive therapeutic targets for asthma.

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Section: Discussionmentioning

confidence: 99%

Section: Erk 1/2 Via Par2mentioning

confidence: 97%

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Polley

Mihara

Ramachandran

et al. 2017

Clin Experimental Allergy

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“…It was discovered that proteolytic cleavage of CCL9 greatly enhances its chemotactic potential [41], suggesting that CCL9 may act as an early response chemokine, responding to local proteolysis that enhances the chemotactic signal. This is of particular interest to our work in that our group has recently demonstrated that HDE is capable of inducing proteolytic cleavage/activation of PAR receptors [42] by HDE. It is therefore possible that CCL9 may be similarly activated in the alveolar space in response to barn dust.…”

Section: Discussionmentioning

confidence: 99%

Effect of low-level CO2 on innate inflammatory protein response to organic dust from swine confinement barns

Schneberger

DeVasure

Bailey

et al. 2017

J Occup Med Toxicol

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BackgroundOrganic hog barn dust (HDE) exposure induces lung inflammation and long-term decreases in lung function in agricultural workers. While concentrations of common gasses in confined animal facilities are well characterized, few studies have been done addressing if exposure to elevated barn gasses impacts the lung immune response to organic dusts. Given the well documented effects of hypercapnia at much higher levels we hypothesized that CO2 at 8 h exposure limit levels (5000 ppm) could alter innate immune responses to HDE.MethodsUsing a mouse model, C57BL/6 mice were nasally instilled with defined barn dust extracts and then housed in an exposure box maintained at one of several CO2 levels for six hours. Bronchiolar lavage (BAL) was tested for several cytokines while lung tissue was saved for mRNA purification and immunohistochemistry.ResultsExposure to elevated CO2 significantly increased the expression of pro-inflammatory markers, IL-6 and KC, in BAL fluid as compared to dust exposure alone. Expression of other pro-inflammatory markers, such as ICAM-1 and matrix metalloproteinase-9 (MMP-9), were also tested and showed similar increased expression upon HDE + CO2 exposure. A chemokine array analysis of BAL fluid revealed that MIP-1γ (CCL9) shows a similar increased response to HDE + CO2. Further testing showed CCL9 was significantly elevated by barn dust and further enhanced by CO2 co-exposure in a dose-dependent manner that was noticeable at the protein and mRNA levels. In all cases, except for ICAM-1, increases in tested markers in the presence of elevated CO2 were only significant in the presence of HDE as well.ConclusionsWe show that even at mandated safe exposure limits, CO2 is capable of enhancing multiple markers of inflammation in response to HDE.

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“…Previously, we detected elastase- and trypsin-like activities in aqueous extracts of poultry dust, and found that heat inactivation at 95 °C for 10 min and serine protease inhibitors such as, α1-antitrypsin and soybean trypsin inhibitor suppressed induction of IL-8 mRNA and protein levels in THP-1 monocytic cells (Bandari, S. K., Master’s Thesis, University of Texas Health Science Center at Tyler, 2012). While our studies were in progress, the presence of proteases in the swine barn dust and their effects on the induction of cytokine expression in lung cells and mouse lung via activation of PAR-1 and PAR-2 were reported [ 38 ]. A number of noteworthy points distinguish our studies from the recent report; our studies have independently provided further evidence for the involvement of protease activities in organic dust from an entirely different source, namely poultry, in the induction of inflammatory gene expression in lung epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

Proteases and oxidant stress control organic dust induction of inflammatory gene expression in lung epithelial cells

et al. 2016

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BackgroundPersistant inflammatory responses to infectious agents and other components in organic dust underlie lung injury and development of respiratory diseases. Organic dust components responsible for eliciting inflammation and the mechanisms by which they cause lung inflammation are not fully understood. We studied the mechanisms by which protease activities in poultry dust extracts and intracellular oxidant stress induce inflammatory gene expression in A549 and Beas2B lung epithelial cells.MethodsThe effects of dust extracts on inflammatory gene expression were analyzed by quantitative polymerase chain reaction (qPCR), enzyme linked immunosorbent (ELISA) and western blot assays. Oxidant stress was probed by dihydroethidium (DHE) labeling, and immunostaining for 4-hydroxynonenal (4-HNE). Effects on interleukin-8 (IL-8) promoter regulation were determined by transient transfection assay.ResultsDust extracts contained trypsin and elastase activities, and activated protease activated receptor (PAR)-1 and -2. Serine protease inhibitors and PAR-1 or PAR-2 knockdown suppressed inflammatory gene induction. Dust extract induction of IL-8 gene expression was associated with increased DHE-fluorescence and 4-HNE staining, and antioxidants suppressed inflammatory gene induction. Protease inhibitors and antioxidants suppressed protein kinase C and NF-κB activation and induction of IL-8 promoter activity in cells exposed to dust extract.ConclusionsOur studies demonstrate that proteases and intracellular oxidants control organic dust induction of inflammatory gene expression in lung epithelial cells. Targeting proteases and oxidant stress may serve as novel approaches for the treatment of organic dust induced lung diseases. This is the first report on the involvement of oxidant stress in the induction of inflammatory gene expression by organic dust.Electronic supplementary materialThe online version of this article (doi:10.1186/s12931-016-0455-z) contains supplementary material, which is available to authorized users.

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Proteases in agricultural dust induce lung inflammation through PAR-1 and PAR-2 activation

Abstract: Romberger DJ, Heires AJ, Nordgren TM, Souder CP, West W, Liu X, Poole JA, Toews ML, Wyatt TA. Proteases in agricultural dust induce lung inflammation through PAR-1 and PAR-2 activation.

Cited by 34 publications

References 45 publications

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Effect of low-level CO2 on innate inflammatory protein response to organic dust from swine confinement barns

Proteases and oxidant stress control organic dust induction of inflammatory gene expression in lung epithelial cells

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