Cysteine-type cathepsins promote the effector phase of acute cutaneous delayed-type hypersensitivity reactions-猫眼课题宝

DOI：

10.7150/thno.31037

发表时间：

2019-05

期刊：

Theranostics

影响因子：

12.4

通讯作者：

中科院分区：

医学1区

文献类型：

作者：

J. Schwenck;A. Maurer;B. Fehrenbacher;R. Mehling;P. Knopf;N. Mucha;D. Haupt;K. Fuchs;C. Griessinger;Daniel Bukala;J. Holstein;M. Schaller;Irene Gonzalez Menendez;K. Ghoreschi;L. Quintanilla‐Martinez;M. Gütschow;S. Laufer;T. Reinheckel;M. Röcken;H. Kalbacher;B. Pichler;M. Kneilling

半胱氨酸型组织蛋白如组织蛋白酶B参与炎症过程的各个步骤，如抗原处理和血管生成。在这里，我们揭示了半胱氨酸型组织蛋白在T细胞驱动的皮肤迟发性超敏反应(DTHR)效应阶段的作用，以及这一作用在治疗性组织蛋白酶B特异性抑制中的意义。方法：用2，4，6-三硝基氯苯(TNCB)致敏2，4，6-三硝基氯苯(TNCB)致敏野生型组织蛋白酶B缺陷(CTSB-/-)和组织蛋白酶Z-/-缺陷(CTZ-/-)小鼠，右耳给予TNCB攻击，建立急慢性皮肤DTHR模型。通过评估耳肿胀反应和组织病理学来评估皮肤DTHR的严重程度。我们对野生型小鼠发炎的耳朵和淋巴结组织以及银屑病患者的活检组织进行了荧光显微镜观察，重点观察了T细胞、B细胞、巨噬细胞、树突状细胞和NK细胞组织蛋白酶B的表达。通过使用蛋白酶激活的底物样探针的光学成像来非侵入性地测定组织蛋白酶活性。通过蛋白水解酶活性部位共价标记和Western blotting验证组织蛋白酶在体外的表达和活性。结果：非侵入性体内光学成像显示炎症的耳朵和急慢性皮肤DTHR的引流淋巴结中有较强的半胱氨酸型组织蛋白酶活性。中性粒细胞、树突状细胞、巨噬细胞、B、T细胞和自然杀伤(NK)细胞表达组织蛋白酶B。在银屑病患者的斑块中也发现了类似的表达模式。生化方法证实组织蛋白酶B在皮肤DTHR小鼠组织中具有活性。局部应用组织蛋白酶B抑制剂可显著减少急性DTHR患者的耳肿胀，但不能减轻慢性DTHR患者的耳肿胀。与野生型小鼠相比，CTSB-/-小鼠在急性DTHR期间表现出增强的耳肿胀反应，尽管缺乏组织蛋白酶B的表达。组织蛋白酶Z是一种与组织蛋白酶B密切相关的蛋白酶，在CTSB-/-小鼠的炎症耳朵中表现出代偿性表达，而在CtsZ-/-小鼠中组织蛋白酶B的表达则相反地升高。结论：组织蛋白酶B积极参与急性皮肤DTHR的效应期。因此，局部应用组织蛋白酶B抑制剂可能有效地限制DTHR，如接触性皮炎或牛皮癣。然而，组织蛋白酶B和Z基因敲除小鼠的实验表明，这两种半胱氨酸型蛋白酶的作用是互补的。

Cysteine-type cathepsins such as cathepsin B are involved in various steps of inflammatory processes such as antigen processing and angiogenesis. Here, we uncovered the role of cysteine-type cathepsins in the effector phase of T cell-driven cutaneous delayed-type hypersensitivity reactions (DTHR) and the implication of this role on therapeutic cathepsin B-specific inhibition. Methods: Wild-type, cathepsin B-deficient (Ctsb-/-) and cathepsin Z-deficient (Ctsz-/-) mice were sensitized with 2,4,6-trinitrochlorobenzene (TNCB) on the abdomen and challenged with TNCB on the right ear to induce acute and chronic cutaneous DTHR. The severity of cutaneous DTHR was assessed by evaluating ear swelling responses and histopathology. We performed fluorescence microscopy on tissue from inflamed ears and lymph nodes of wild-type mice, as well as on biopsies from psoriasis patients, focusing on cathepsin B expression by T cells, B cells, macrophages, dendritic cells and NK cells. Cathepsin activity was determined noninvasively by optical imaging employing protease-activated substrate-like probes. Cathepsin expression and activity were validated ex vivo by covalent active site labeling of proteases and Western blotting. Results: Noninvasive in vivo optical imaging revealed strong cysteine-type cathepsin activity in inflamed ears and draining lymph nodes in acute and chronic cutaneous DTHR. In inflamed ears and draining lymph nodes, cathepsin B was expressed by neutrophils, dendritic cells, macrophages, B, T and natural killer (NK) cells. Similar expression patterns were found in psoriatic plaques of patients. The biochemical methods confirmed active cathepsin B in tissues of mice with cutaneous DTHR. Topically applied cathepsin B inhibitors significantly reduced ear swelling in acute but not chronic DTHR. Compared with wild-type mice, Ctsb-/- mice exhibited an enhanced ear swelling response during acute DTHR despite a lack of cathepsin B expression. Cathepsin Z, a protease closely related to cathepsin B, revealed compensatory expression in inflamed ears of Ctsb-/- mice, while cathepsin B expression was reciprocally elevated in Ctsz-/- mice. Conclusion: Cathepsin B is actively involved in the effector phase of acute cutaneous DTHR. Thus, topically applied cathepsin B inhibitors might effectively limit DTHR such as contact dermatitis or psoriasis. However, the cathepsin B and Z knockout mouse experiments suggested a complementary role for these two cysteine-type proteases.

Cysteine-type cathepsins promote the effector phase of acute cutaneous delayed-type hypersensitivity reactions

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