Innate function of house dust mite allergens: robust enzymatic degradation of extracellular matrix at elevated pH
Innate function of house dust mite allergens: robust enzymatic degradation of extracellular matrix at elevated pH
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DOI：
10.1186/s40413-017-0154-3
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发表时间：
2017-07-04
影响因子：
5.1
通讯作者：
Jensen-Jarolim, Erika
中科院分区：
文献类型：
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作者：
Oida, Kumiko;Einhorn, Lukas;Jensen-Jarolim, Erika
背景：接触屋尘螨（Dermatophagoides pteronyssinus，D. p.）增加了人类和他们最好的朋友狗患过敏性疾病的风险。在这里，我们探讨了这种过敏性螨通过其酶是否可能影响皮肤细胞外基质（ECM），这在结构上和functional.Methods的关键决定上皮屏障的完整性：从灰尘纯化或培养的D. P.机构获得的两种提取物在本研究中使用。为了评估D. p.对ECM蛋白组分的潜在影响，通过酪蛋白和明胶凝胶酶谱法测定D. p.提取物的蛋白水解活性，并通过比色法测定其N-乙酰基-β-氨基己糖苷酶活性。此外，IgE依赖性和先天性脱粒潜力的D. P。在犬MPT-1肥大细胞和神经突生长测定使用大鼠嗜铬细胞瘤PC-12 cells.Results：在凝胶zymography，这两种提取物消化底物酪蛋白和明胶的剂量依赖性的方式，特别是在碱性pH值，并在很宽的温度范围内（30摄氏度-42摄氏度）有效。特别是，一个25 kDa的带对应于Der p 1，主要的D. p.过敏原的人，被发现在酪蛋白和明胶凝胶酶活性，无论金属离子和碱性条件的存在。除了蛋白酶活性外，在两种提取物中均检测到N-乙酰基-β-氨基己糖苷酶活性，这表明D. p.通过使蛋白质和糖胺聚糖降解来影响皮肤ECM。虽然这两个D. P.提取物诱导IgE依赖性肥大细胞脱颗粒，少得多的先天性肥大细胞和神经元cells observed.Conclusions的影响：我们的数据强调，D. P.是一个强大的来源，几个不同的酶与蛋白酶和N-乙酰-β-氨基己糖苷酶的活动。在碱性环境中，它们可以降解ECM的组分。因此，D. p.可能导致上皮屏障破坏，尤其是当皮肤表面pH升高时。
Background: Exposure to the house dust mite Dermatophagoides pteronyssinus (D.p.) increases the risk for developing allergic diseases in humans and their best friends, the dogs. Here, we explored whether this allergenic mite via its enzymes may impact the cutaneous extracellular matrix (ECM), which critically determines epithelial barrier integrity both structurally and functionally.Methods: Two extracts obtained from either dust-purified or cultured D.p. bodies were used in the present study. To assess the potential impact of D.p. on protein components of the ECM, proteolytic activity of the D.p. extracts were determined by casein and gelatin gel zymography, and their N-acetyl-beta-hexosaminidase activity determined colorimetrically. In addition, IgE-dependent and innate degranulation potential of D.p. was examined in canine MPT-1 mast cells and neurite outgrowth assay using rat pheochromocytoma PC-12 cells.Results: In gel zymography, both extracts digested the substrates casein and gelatin in a dose-dependent manner, especially at alkaline pH, and effective in a wide range of temperatures (30 degrees C-42 degrees C). In particular, a 25-kDa band corresponding to Der p 1, the major D.p. allergen for humans, was found enzymatically active in both casein and gelatin gels regardless of the presence of metal ions and of alkaline conditions. Besides protease activity, N-acetyl-beta-hexosaminidase activity was detected in both extracts, suggesting that D.p. affects the cutaneous ECM through deteriorating both proteins and glycosaminoglycans. While both D.p. extracts induced IgE-dependent mast cell degranulation, much less innate effects on mast-and neuronal cells were observed.Conclusions: Our data highlight that D.p. is a robust source of several distinct enzymes with protease-and N-acetyl-beta-hexosaminidase activities. In alkaline milieu they can degrade components of the ECM. Therefore, D.p. may contribute to epithelial barrier disruption especially when the skin surface pH is elevated.