Investigating the role of a Tannerella forsythia HtrA protease in host protein degradation and inflammatory response

S. Bloch, Fiona F. Hager-Mair, Johanna Bacher, Markus B. Tomek, Bettina Janesch, O. Andrukhov, Christina Schäffer
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Abstract

Degradation of host proteins by bacterial proteases leads to the subversion of the host response and disruption of oral epithelial integrity, which is considered an essential factor in the progression of periodontitis. High-temperature requirement A (HtrA) protease, which is critical for bacterial survival and environmental adaptation, is found in several oral bacteria, including the periodontal pathogen Tannerella forsythia. This study investigated the proteolytic activity of HtrA from T. forsythia and its ability to modulate the host response.HtrA of T. forsythia was identified bioinformatically and produced as a recombinant protein. T. forsythia mutants with depleted and restored HtrA production were constructed. The effect of T. forsythia wild-type, mutants and recombinant HtrA on the degradation of casein and E-cadherin was tested in vitro. Additionally, the responses of human gingival fibroblasts and U937 macrophages to the different HtrA-stimuli were investigated and compared to those triggered by the HtrA-deficient mutant.T. forsythia wild-type producing HtrA as well as the recombinant enzyme exhibited proteolytic activity towards casein and E-cadherin. No cytotoxic effect of either the wild-type, T. forsythia mutants or rHtrA on the viability of host cells was found. In hGFB and U937 macrophages, both T. forsythia species induced an inflammatory response of similar magnitude, as indicated by gene and protein expression of interleukin (IL)-1β, IL-6, IL-8, tumour necrosis factor α and monocyte chemoattractant protein (MCP)-1. Recombinant HtrA had no significant effect on the inflammatory response in hGFBs, whereas in U937 macrophages, it induced a transient inflammatory response at the early stage of infection.HtrA of T. forsythia exhibit proteolytic activity towards the host adhesion molecule E-cadherin and has the potential to influence the host response. Its role in the progression of periodontitis needs further clarification.
研究连翘炭疽杆菌 HtrA 蛋白酶在宿主蛋白质降解和炎症反应中的作用
细菌蛋白酶对宿主蛋白质的降解导致宿主反应的颠覆和口腔上皮完整性的破坏,这被认为是牙周炎进展的一个重要因素。高温要求 A(HtrA)蛋白酶对细菌的生存和环境适应至关重要,它存在于多种口腔细菌中,包括牙周致病菌连翘担子菌。本研究调查了连翘担子菌的 HtrA 蛋白水解活性及其调节宿主反应的能力。通过生物信息学方法鉴定了连翘担子菌的 HtrA,并将其制成重组蛋白。通过生物信息学方法鉴定了连翘HtrA,并将其制成重组蛋白。体外测试了连翘野生型、突变体和重组 HtrA 对酪蛋白和 E-cadherin 降解的影响。此外,还研究了人牙龈成纤维细胞和 U937 巨噬细胞对不同 HtrA 刺激的反应,并与 HtrA 缺陷突变体引发的反应进行了比较。连翘野生型产生的 HtrA 和重组酶对酪蛋白和 E-cadherin 具有蛋白水解活性。野生型、连翘突变体或 rHtrA 对宿主细胞的活力都没有细胞毒性作用。在 hGFB 和 U937 巨噬细胞中,两种连翘菌诱导的炎症反应程度相似,白细胞介素(IL)-1β、IL-6、IL-8、肿瘤坏死因子 α 和单核细胞趋化蛋白(MCP)-1 的基因和蛋白表达均表明了这一点。重组 HtrA 对 hGFBs 中的炎症反应无明显影响,而在 U937 巨噬细胞中,它能在感染早期诱发短暂的炎症反应。它在牙周炎进展过程中的作用有待进一步明确。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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