Normal And Pathologic Mechanisms Of Inflammation, Innate And Acquired Immunity

自身免疫和干燥综合征（SS）（55%） 我们研究的一个重要目的是确定炎症和免疫反应出错的潜在机制，导致自身免疫发病机制，如干燥综合征。SS的自身免疫标志包括外分泌腺的局灶性淋巴细胞浸润（病灶评分）和自身抗体的产生。SS的病因在很大程度上仍然未知，并且许多对其他自身免疫性疾病有效的药物，特别是TNF拮抗剂的失败提出了一个难题。TNF抑制剂缺乏疗效的一个潜在线索是SS患者外周循环中IL-17升高，这也不受抗TNF治疗的调节。在唾液腺活检中，我们进行了基因分析，以将特定基因表达与炎症组织中的T细胞亚群和/或其产物相关联。在细胞水平，我们发现IL-17和TGF β的表达随着活检病灶评分的升高而逐渐增加，将IL-17与SS组织病理学联系起来。局部涉及的先天免疫防御群体包括巨噬细胞，在我们对SS组织的微阵列分析中，我们专注于疾病相关的过表达基因。 其中最高表达的基因是哺乳动物几丁质酶家族的成员，以前与外分泌蛋白病无关。几丁质酶-3-like-1（CHI 3L 1/YKL-40）和几丁质酶1（CHIT 1）是哺乳动物几丁质酶样糖蛋白家族的高度保守成员，在转录组水平上是明显的，并且在发炎的唾液腺中被检测为糖蛋白。由于这些和其他巨噬细胞衍生分子的表达升高对应于SS中更严重的疾病，因此这些观察结果暗示了潜在的免疫病理巨噬细胞参与，此外，通过口服和/或全身检测的几丁质酶和/或其他巨噬细胞产物可提供诊断工具。 SLPI调节炎症和癌症中的蛋白水解级联反应（30%） 在没有SLPI的情况下，增强的炎症、延迟愈合和对感染的易感性表明SLPI和炎症反应的调节之间存在联系。基于一系列的实验，我们证明了SLPI-膜联蛋白II相互作用中断负责纤溶酶生成的蛋白水解级联。此外，我们已经表明，在SLPI缺失小鼠中，巨噬细胞处理不受调节的纤溶酶水平，这有助于过度的组织降解，增加炎症和组织损伤后的愈合失败。通过抑制纤溶酶（其具有与补体活化、纤维蛋白溶解、细胞因子调节、TGF-β活化和其它炎性后遗症相关的无数下游后果），SLPI可以对蛋白水解和炎性活化网络施加控制。由于纤维蛋白溶解过程不仅是炎症、凝血和组织修复的基础，而且越来越多的证据表明其在多种慢性炎症性疾病、心血管疾病和恶性肿瘤的发病机制中的失调，因此进一步定义和调节这些途径具有重要意义。 与这些发现相关，我们与UMD的研究人员合作，探讨了SLPI在转移性头颈部鳞状细胞癌（HNSCC）中的作用，HNSCC仍然是全球10种最常见的癌症之一，预后非常差。SLPI在非浸润性组织中的表达高于恶性组织，且表达水平与HNSCC的分期和浸润方式呈显著负相关。此外，我们的新数据表明肿瘤细胞因子对这些侵袭性途径的调节，这表明了一种调节口腔癌肿瘤侵袭潜力的策略。 TNF α作为异常皮肤伤口愈合的治疗靶点（15%） 受损的伤口愈合状态导致大量的发病率和治疗成本，导致美国每年数十亿美元的支出。慢性伤口和受损的急性伤口都以过度炎症、增强的蛋白水解和减少的基质沉积为特征。这些混杂因素在老年人中加重，部分与局部和全身肿瘤坏死因子α（TNF）水平升高有关。此外，我们已经使用了严重受损的伤口愈合和过度炎症的SLPI无效小鼠模型，与年龄相关的延迟人类愈合相当，以证明局部应用抗TNF中和抗体减弱白细胞募集和NF B活化，改变M1和M2巨噬细胞之间的平衡，并加速伤口愈合。在TNF拮抗作用后，基质合成增强，与炎症参数和NF B结合活性的抑制相关。我们的数据表明，抑制TNF是逆转与缺乏SLPI相关的严重受损的愈合反应的关键事件，并且可能适用于预防和/或治疗人类受损的伤口愈合状态。

Autoimmunity and Sjogren's syndrome (SS) (55%) An important objective of our studies is to define the underlying mechanisms by which inflammatory and immune responses go awry, resulting in autoimmune pathogenesis, as exemplified by Sjogren's syndrome. The autoimmune hallmarks of SS include focal lymphoid cell infiltration of exocrine glands (focus score) and production of autoantibodies. The etiology of SS remains largely unknown and a conundrum is presented by the failure of many drugs that are effective in other autoimmune disorders, particularly TNF antagonists. A potential clue to the lack of efficacy of TNF inhibitors is the elevated IL-17 in SS patients peripheral circulation, which is also unmodulated by anti-TNF therapy. In salivary gland biopsies, we performed gene profiling to correlate specific gene expression with T cell subsets and/or their products within the inflamed tissues. At the cellular level, we showed that expression of IL-17 and also TGFbeta progressively increases with higher biopsy focus scores, connecting IL-17 to SS tissue pathology. Locally involved innate immune defense populations include macrophages, and in our microarray analysis of SS tissues, we focused on disease related overexpressed genes. Among the most highly expressed genes were members of the mammalian chitinase family, not previously associated with exocrinopathies. Both chitinase-3-like-1 (CHI3L1/YKL-40) and chitinase 1 (CHIT1), highly conserved members of the mammalian chitinase-like glycoprotein family were evident at the transcriptome levels, as well as detected as glycoproteins within inflamed salivary glands. Since the elevated expression of these and other macrophage-derived molecules corresponded with more severe disease in SS, these observations implicate a potential immunopathologic macrophage involvement and furthermore, that chitinases and/or other macrophage products detected orally and/or systemically may provide a diagnostic tool. SLPI regulation of proteolytic cascade in inflammation and cancer (30%) Enhanced inflammation, delayed healing and susceptibility to infection in the absence of SLPI have suggested a link between SLPI and regulation of inflammatory responses. Based on a series of experiments, we demonstrated that a SLPI-annexin II interaction interrupts the proteolytic cascade responsible for plasmin generation. Moreover, we have shown that in SLPI null mice, macrophages process unregulated levels of plasmin, which contributes to excess tissue degradation, increased inflammation and failed healing following tissue injury. By inhibiting plasmin, which has a myriad of downstream consequences relative to complement activation, fibrinolysis, cytokine regulation, TGF-beta activation and other inflammatory sequelae, SLPI can exert control over proteolytic and inflammatory activation networks. Because fibrinolytic processes are fundamental not only to inflammation, coagulation and tissue repair, but growing evidence highlights their dysregulation in the pathogenesis of multiple chronic inflammatory disorders, cardiovascular disease and malignancy, further defining and regulating these pathways is of major significance. Relevant to these findings, we have explored the role of SLPI in metastatic head and neck squamous cell carcinoma (HNSCC), which remains one of the 10 most frequently occurring cancers worldwide with a very poor prognosis, in collaboration with investigators at UMD. SLPI expression was higher in noninvolved than in malignant tissue, and levels significantly inversely correlated with HNSCC stage and pattern of invasion. Moreover, our new data demonstrate tumor cytokine regulation of these invasive pathways suggesting a strategy to modulate tumor invasive potential in oral cancer. TNFalpha as a therapeutic target for aberrant cutaneous wound healing (15%) Impaired wound healing states lead to substantial morbidity and cost with treatment resulting in an expenditure of billions of dollars per annum in the USA. Both chronic wounds and impaired acute wounds are characterized by excessive inflammation, enhanced proteolysis, and reduced matrix deposition. These confounding factors are exacerbated in the elderly, in part, related to increased local and systemic tumor necrosis factor alpha(TNFα) levels. Moreover, we have used SLPI null mouse model of severely impaired wound healing and excessive inflammation, comparable to age-related delayed human healing, to demonstrate that topical application of anti-TNFα neutralizing antibodies blunts leukocyte recruitment and NFκB activation, alters the balance between M1 and M2 macrophages, and accelerates wound healing. Following antagonism of TNFα, matrix synthesis is enhanced, associated with suppression of both inflammatory parameters and NFκB binding activity. Our data suggest that inhibiting TNFα is a critical event in reversing the severely impaired healing response associated with the absence of SLPI, and may be applicable to prophylaxis and/or treatment of impaired wound healing states in humans.