Presence of the house dust mite allergen in the gastrointestinal tract of dogs with chronic enteropathy: A potential inducer of interleukin-1β

Takahashi, Kaho; Yanuma, Nanako; Hirokawa, Marin; Miyajima, Masaki; Ogawa, Misato; Osada, Hironari; Sugita, Koji; Kondo, Hirotaka; Ohmori, Kitaro

doi:10.1016/j.vetimm.2020.110150

Cited by 6 publications

(11 citation statements)

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“…Der f was preincubated with 10 µg/mL E64 or 1 mM AEBSF for 15 min at 37°C according to previous studies. 7,8 CPEK cells also were incubated with medium alone or 3 nM trypsin (Sigma-Aldrich) as a serine protease for 6, 12 or 24 h at 37°C. Effects of lipopolysaccharide (LPS) present in Der f on IL-33 mRNA expression also were examined in CPEK cells (see Supporting information Appendix S1).…”

Section: Methodsmentioning

confidence: 99%

“…To determine the effects of Der f‐derived proteases on IL‐33 mRNA expression, CPEK cells were incubated for 12 h at 37°C with medium alone, 100 µg/mL Der f, Der f pre‐treated with E64 (Sigma‐Aldrich; St Louis, MO, USA) as a cysteine protease inhibitor, or Der f pre‐treated with 4‐(2‐aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF, Santa Cruz Biotechnology; Dallas, TX, USA) as a serine protease inhibitor. Der f was pre‐incubated with 10 µg/mL E64 or 1 mM AEBSF for 15 min at 37°C according to previous studies 7,8 . CPEK cells also were incubated with medium alone or 3 nM trypsin (Sigma‐Aldrich) as a serine protease for 6, 12 or 24 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…Total RNA extraction, cDNA synthesis and real‐time PCR analysis were performed as described previously 7 . As reference genes, ribosomal protein L13a ( RPL13A ) and succinate dehydrogenase complex flavoprotein subunit A ( SDHA ) were used (Table 1).…”

Section: Methodsmentioning

confidence: 99%

“…Total RNA extraction, cDNA synthesis and real-time PCR analysis were performed as described previously. 7 As reference genes, ribosomal protein L13a (RPL13A) and succinate dehydrogenase complex flavoprotein subunit A (SDHA) were used (Table 1). 2,10 The geometric mean of the cycle threshold (Ct) values of the two reference genes was subtracted from the Ct values of IL-33 (DCT).…”

Section: Real-time Pcrmentioning

confidence: 99%

“…The relative mRNA expression of IL-33 was calculated by the 2 ÀDDCT method. 7 Each value was presented as an n-fold change in mRNA expression relative to the control group.…”

Section: Real-time Pcrmentioning

confidence: 99%

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House dust mite‐derived serine protease upregulates gene expression of interleukin‐33 in canine keratinocytes via protease‐activated receptor‐2

Nishimura

Miyajima

Takahashi

et al. 2021

Veterinary Dermatology

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Background -The involvement of interleukin (IL)-33 produced by keratinocytes has been suggested in the pathogenesis of canine atopic dermatitis (cAD). House dust mite (HDM)-derived proteases induce the production of various cytokines and chemokines in keratinocytes via protease-activated receptor-2 (PAR-2); however, their effects on IL-33 mRNA expression in canine keratinocytes have not been determined.Hypothesis/Objective -To clarify whether HDM-derived proteases induce IL-33 mRNA expression in canine keratinocytes via PAR-2.Methods and materials -Expression of IL-33 mRNA was quantified by real-time PCR in a cell line of canine progenitor epidermal keratinocytes (CPEK) stimulated with Dermatophagoides farinae (Der f) whole body extract, Der f pre-treated with cysteine protease and serine protease inhibitors, and trypsin. Trypsin and Der f-mediated IL-33 mRNA expression also was measured in CPEK cells treated with a PAR-2 antagonist.Results -Der f enhanced IL-33 mRNA expression in CPEK cells in incubation time-and dose-dependent manners. Der f pre-treated with a serine protease inhibitor, and not a cysteine protease inhibitor, abrogated an increase in IL-33 mRNA expression in CPEK cells. Trypsin also enhanced IL-33 mRNA expression in CPEK cells. Trypsin-mediated IL-33 mRNA expression was completely abolished by a PAR-2 antagonist, while Der fmediated IL-33 mRNA expression was partially and significantly diminished by it.Conclusions and clinical relevance -Der f-derived serine protease upregulated IL-33 mRNA expression in CPEK cells at least in part via PAR-2. These findings suggest that HDM may be involved in the development of C AD by increasing IL-33 mRNA expression in keratinocytes.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Real-time Pcrmentioning

confidence: 99%

“…The relative mRNA expression of IL-33 was calculated by the 2 ÀDDCT method. 7 Each value was presented as an n-fold change in mRNA expression relative to the control group.…”

Section: Real-time Pcrmentioning

confidence: 99%

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