Targeting professional APCs using Fasciola hepatica FABP to suppresses inflammation

使用肝片形吸虫 FABP 靶向专业 APC 抑制炎症

基本信息

批准号：
10670212
负责人：
ANA M ESPINO
金额：
$ 37.5万
依托单位：
UNIVERSITY OF PUERTO RICO MED SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10670212
关键词：
Animals Anti-Inflammatory Agents Antigen-Presenting Cells Antiinflammatory Effect Antioxidants Attenuated Binding Biological Assay CD14 gene CD4 Positive T Lymphocytes CD80 gene CD86 gene Categories Cell Culture Techniques Cell surface Clinical Coculture Techniques Colitis Colon Data Dendritic Cells Dendritic cell activation Development Disease Docking Domestic Animals Dose Drug Targeting Endotoxemia Experimental Autoimmune Encephalomyelitis Exposure to FOXP3 gene Fasciola hepatica Fascioliasis Flow Cytometry Foundations Genes Genetic Transcription Goals Greater sac of peritoneum Health Helminth Antigens Helminths Hepatica Human IL2RA gene IL6 gene Immune Immune response Immune system Immunosuppression In Vitro Inbred NOD Mice Infection Inflammation Inflammatory Inflammatory Bowel Diseases Inflammatory Response Injections Insulin-Dependent Diabetes Mellitus Interleukin-1 beta Interleukin-10 Knockout Mice Ligands Ligation Macrophage Measures Medical Modeling Multiple Sclerosis Mus Myelogenous Natural Immunity Nature Parasites Parasitic infection Pathologic Peritoneal Macrophages Pharmaceutical Preparations Play Population Production Property Protein Family Proteins Proteomics Recombinants Reporting Research Role Schistosomiasis Sepsis Septic Shock Serum Signal Pathway Signal Transduction Specificity Structure Symptoms TLR2 gene TLR4 gene TNFRSF5 gene Testing Therapeutic Therapeutic Effect Time Treatment Efficacy Ulcerative Colitis Variant Work animal tissue antagonist base chemokine chronic infection cytokine cytokine release syndrome dextran sulfate sodium induced colitis drug development efficacy testing experimental study fatty acid-binding proteins gut inflammation helminth infection immunoregulation in vivo innovation lipid metabolism mortality mouse model novel novel strategies prevent receptor response septic synthetic polymer Bioplex vaccine candidate

项目摘要

ABSTRACT The growing use of `helminth-therapy' in the last decade to prevent or ameliorate inflammatory diseases is a promising and novel approach. Fasciola hepatica, one of the most globally prevalent parasitic helminths of domestic animals, is particularly adept at controlling the immune response of its host. At early stages of infection the parasite induces a dominant Th2/T-regulatory-type immune response coincidental with suppression of Th1 responses. Recent studies have demonstrated that F. hepatica infection attenuates the clinical signs of murine experimental autoimmune encephalomyelitis [1] and prevents the development of Type-1 diabetes in a non-obese diabetic mouse model [2]. However, the immunoregulation associated to F. hepatica lacks specificity and results in a compromised immune system unable to respond effectively to bystander infections [3, 4]. A better alternative for drug development would be to identify parasite molecules with immune-modulatory capacity and to characterize their precise mechanism of action. Fatty acid binding proteins (FABPs) are proteins that play an important role in the parasite's lipid metabolism and have been recently categorized as anti-oxidant molecules [5]. We have demonstrated that a recombinant 14.5kDa protein belonging to the fatty acid binding protein (Fh15) is able to significantly suppress the cytokine storm when is applied therapeutically 1h after exposure to lethal doses of LPS, which suggest that Fh15 could act as a TLR4-antagonist. Moreover, we also found that Fh15-treatment increased accumulation of large and small peritoneal MΦs (LPMs and SPMs) into the peritoneal cavity (PerC) compared to the of septic mice [6]. Moreover, we also demonstrated by proximity ligation assay (PLA) that the native F. hepatica FABP variant (Fh12) binds to the human CD14-coreceptor [7], which has been associated to the activation-signaling cascade of various TLRs [8-11]. Because TLR4 is directly involved in the inflammatory responses during sepsis and ulcerative colitis (UC), this role for Fh15 could have a significant impact on human health [12, 13]. Our central hypothesis is that the suppression of pro-inflammatory cytokines induced by injection (i.p.) with Fh15 is enough to prevent the high mortality in mice exposed to a lethal dose of LPS and to prevent or ameliorate the intestinal inflammation in a DSS-induced mouse model of UC. Both therapeutic effects will be directly associated with the production of M2-type MΦs in the PerC of animals and the induction of tolerogenic properties of DCs, which will be critically dependent on CD14. The proposed research is conceptually innovative because it is the first time that a protein of the F. hepatica FABP family, specifically Fh15, is investigated as an anti-inflammatory molecule and there are no reported works on MΦs and DCs in relation to Fh15 exposure in vivo or in vitro. Understanding how Fh15 modulate and interact with MΦs and DCs at early stages of innate immunity could contribute to the development of anti-inflammatory drugs against sepsis and other inflammatory diseases. The specific aims are: 1) Determine the capacity of Fh15 to suppress the pathologic effects of septic shock and ulcerous colitis (UC) in the mouse model and, 2) Determine whether Fh15 induces the tolerogenic properties of DCs and identify mechanism- underlying immunosuppression caused by Fh15. The proposed studies significantly will advance anti- inflammatory drug development based on helminth-antigen derived therapy by providing a well-defined F. hepatica molecule as drug target mechanism of action.

抽象的过去十年中越来越多地使用“蠕虫疗法”来预防或改善炎症性疾病肝片形吸虫是全球最流行的国内寄生蠕虫之一，前景广阔。动物，特别擅长在感染寄生虫的早期阶段控制宿主的免疫反应。诱导显性 Th2/T 调节型免疫反应，同时抑制 Th1 反应。最近的研究表明，肝镰刀菌感染可减弱小鼠实验性肝镰刀菌的临床症状。自身免疫性脑脊髓炎 [1] 并预防非肥胖糖尿病小鼠发展为 1 型糖尿病模型 [2] 然而，与 F. hepatica 相关的免疫调节缺乏特异性，导致功能受损。免疫系统无法有效应对旁观者感染 [3, 4]。开发将是识别具有免疫调节能力的寄生虫分子并表征其脂肪酸结合蛋白（FABP）是在脂肪酸结合蛋白中发挥重要作用的蛋白质。寄生虫的脂质代谢，最近被归类为抗氧化分子 [5]。证明属于脂肪酸结合蛋白 (Fh15) 的重组 14.5kDa 蛋白能够暴露于致死剂量的 LPS 1 小时后进行治疗，可显着抑制细胞因子风暴，这表明Fh15可以作为TLR4拮抗剂，此外，我们还发现Fh15治疗增加了。与腹膜腔（PerC）相比，大和小的腹膜 MΦ（LPM 和 SPM）的积累此外，我们还通过邻近连接试验（PLA）证明了天然肝片杆菌。 FABP 变体 (Fh12) 与人类 CD14 辅助受体结合 [7]，该受体与激活信号传导相关因为 TLR4 直接参与脓毒症和脓毒症期间的炎症反应。溃疡性结肠炎 (UC) 中，Fh15 的这种作用可能对人类健康产生重大影响 [12, 13]。假设是，通过注射 (i.p.) Fh15 诱导的促炎细胞因子的抑制足以防止暴露于致死剂量 LPS 的小鼠的高死亡率，并预防或改善肠道 DSS 诱导的 UC 小鼠模型中的炎症这两种治疗效果都与在动物的 PerC 中产生 M2 型 MΦ 并诱导 DC 的耐受性，这将是所提出的研究在概念上是创新的，因为这是第一次 F. hepatica FABP 家族的蛋白质，特别是 Fh15，被研究作为一种抗炎分子，并且有目前还没有关于 MΦ 和 DC 与 Fh15 体内或体外暴露相关的研究报道。在先天免疫的早期阶段调节 MΦ 和 DC 并与之相互作用可能有助于抗脓毒症和其他炎症性疾病的抗炎药物的具体目标是：1）确定。 Fh15 抑制小鼠感染性休克和溃疡性结肠炎 (UC) 病理效应的能力模型，2) 确定 Fh15 是否诱导 DC 的耐受性并确定机制 - Fh15 引起的潜在免疫抑制。拟议的研究将显着推进抗-免疫抑制。通过提供明确的肝蠕虫抗原衍生疗法来开发炎症药物分子作为药物靶点的作用机制。