Anti-Inflammatory mechanisms of soybean-derived Bowman-Birk protease inhibitor

描述（由申请人提供）：Bowman-Birk Inhibitor（BBI）是一种来自大豆的口服活性丝氨酸蛋白酶抑制剂。BBI浓缩物（BBIC）是富含BBI的大豆提取物。在非神经性自身免疫性疾病的人体试验中，BBIC已显示出有效的抗炎作用，而没有相关的毒性。我们已经表明，在几种啮齿动物模型中，口服BBI/BBIC显著降低了中枢神经系统的自身免疫性炎症。重要的是，在疾病发作后，当临床缺陷已经发展时，施用BBI迅速改善临床体征并防止疾病进展。BBI疗法的这一特点是其适用于治疗已经显示出疾病临床体征的患者的核心。尽管BBIC已经过临床测试，但其抗炎作用的机制在神经和非神经自身免疫性疾病中的了解都很少。在这里，我们建议调查的影响，BBI在体内和体外免疫细胞使用的小鼠模型的多发性硬化症（MS）主要由CD 4 +T细胞介导的，和糖尿病模型主要由CD 8 +T细胞介导的。在具体目标1中，我们将检验IL-10产生的增加是BBI在EAE中抗炎作用的重要机制的假设。这一假设是基于我们的发现，即BBI诱导体内和体外免疫细胞产生IL-10。已经显示BBI抑制抗原加工，但其对抗原呈递细胞功能的影响尚不清楚。在目标2中，我们将检验BBI抑制抗原递呈的假设。到目前为止，还没有研究表明BBI对T细胞的直接影响。考虑到T细胞在适应性免疫中的关键作用，在目标3中，我们将测试BBI直接抑制T细胞的假设。我们期望这些研究将深入了解BBI抗炎作用的机制方面，并促进其作为MS以及其他炎症性疾病的补充治疗。 Bowman-Birk Inhibitor（BBI）是一种来自大豆的口服活性蛋白酶抑制剂。在自身免疫性疾病的人体试验中，BBI已被证明具有抗炎作用，而没有相关的毒性。尽管BBI已经进行了临床试验，但其抗炎作用的机制仍知之甚少。在这里，我们建议研究BBI对多发性硬化症和糖尿病实验模型中免疫细胞的影响。在我们以前的研究中，我们发现BBI增加白细胞介素10（IL-10）的产生。IL-10具有众所周知的抗炎特性，我们将研究这种分子是否在BBI的有益作用中发挥作用。已发表的数据和初步研究表明，BBI对两种关键类型的免疫细胞有影响;所谓的抗原呈递细胞和T细胞。该提案的第二部分致力于BBI对这些细胞的影响的机制研究。我们相信，这些研究将提供深入了解BBI的抗炎作用，并促进其作为炎症性疾病的辅助治疗。

DESCRIPTION (provided by applicant): Bowman-Birk Inhibitor (BBI) is an orally active serine protease inhibitor derived from soybeans. BBI concentrate (BBIC) is an extract of soybeans enriched in BBI. In human trials for non-neurological autoimmune disorders, BBIC has been shown to exert potent anti-inflammatory effects without associated toxicity. We have shown that in several rodent models oral BBI/BBIC dramatically reduced autoimmune inflammation of the central nervous system. It is of major significance that after disease onset, when clinical deficits have already developed, administration of BBI rapidly ameliorated clinical signs and prevented disease progression. This feature of BBI therapy is central to its suitability for treating patients who have already shown clinical signs of disease. Even though BBIC has been clinically tested, mechanisms of its anti-inflammatory action are poorly understood in both neurological and non-neurological autoimmune diseases. Here we propose to investigate effects of BBI on immune cells in vivo and in vitro using a mouse model of multiple sclerosis (MS) mediated predominantly by CD4+T cells, and a model of diabetes mediated predominantly by CD8+T cells. In specific aim 1 we will test the hypothesis that increase in IL-10 production is an important mechanism for anti-inflammatory effects of BBI in EAE. This hypothesis is based on our discovery that BBI induces IL-10 production by immune cells in vivo and in vitro. It has been shown that BBI inhibits antigen processing, but its effect on functions of antigen presenting cells is unknown. In aim 2 we will test the hypothesis that BBI suppresses antigen presentation. Thus far, there is no study that addresses direct effects of BBI on T cells. Taking in account the crucial role of T cells in adaptive immunity, in aim 3 we will test the hypothesis that BBI directly suppresses T cells. We expect that these studies will yield in depth insights into mechanistic aspects of anti-inflammatory actions of BBI, and facilitate its use as a supplementary treatment for MS, as well as other inflammatory conditions. Public Health Relevance: Bowman-Birk Inhibitor (BBI) is an orally active protease inhibitor from soybeans. In human trials for autoimmune disorders, BBI has been shown to have anti-inflammatory effects without associated toxicity. Even though BBI has already been clinically tested, mechanisms of its anti-inflammatory action are poorly understood. Here we propose to investigate effects of BBI on immune cells in experimental models of multiple sclerosis and diabetes. In our previous study we discovered that BBI increases production of interleukin 10 (IL-10). IL-10 has well known anti-inflammatory properties, and we will investigate if this molecule plays a role in beneficial effects of BBI. Published data and preliminary studies indicate that BBI has an effect on two crucial types of immune cells; so called antigen presenting cells and T cells. The second portion of this proposal is dedicated to mechanistic studies of effects that BBI has on these cells. We believe that these studies will provide in depth insights into anti-inflammatory actions of BBI, and facilitate its use as a supplementary treatment for inflammatory diseases.