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

描述(申请人提供)：鲍曼-伯克抑制剂(BBI)是一种从大豆中提取的具有口服活性的丝氨酸蛋白酶抑制剂。BBI浓缩物(BBIC)是一种富含BBI的大豆提取物。在非神经系统自身免疫性疾病的人体试验中，BBIC已被证明具有强大的抗炎作用，且没有相关毒性。我们已经证明，在几种啮齿动物模型中，口服BBI/BBIC显着减少了中枢神经系统的自身免疫性炎症。在疾病发生后，当临床缺陷已经形成时，应用BBI迅速改善临床症状并防止疾病进展，这一点具有重要意义。BBI疗法的这一特点是其适用于治疗已经表现出临床症状的患者的核心。尽管BBIC已经过临床测试，但其抗炎作用机制在神经系统和非神经系统自身免疫性疾病中都知之甚少。在此，我们建议使用以CD4+T细胞为主的多发性硬化症(MS)小鼠模型和以CD8+T细胞为主的糖尿病模型来研究BBI对体内和体外免疫细胞的影响。在特定的目标1中，我们将检验这一假设，即增加IL-10的产生是BBI在EAE中抗炎作用的重要机制。这一假说是基于我们的发现，即BBI在体内和体外诱导免疫细胞产生IL-10。已有研究表明，BBI可抑制抗原加工，但其对抗原提呈细胞功能的影响尚不清楚。在目标2中，我们将检验BBI抑制抗原提呈的假设。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细胞在获得性免疫中的关键作用，在目标3中，我们将检验BBI直接抑制T细胞的假设。我们期待这些研究将对BBI抗炎作用的机制方面产生深入的见解，并促进其作为MS的辅助治疗以及其他炎症条件的使用。 与公共卫生相关：鲍曼-伯克抑制剂(BBI)是一种来自大豆的口服活性蛋白酶抑制剂。在针对自身免疫性疾病的人体试验中，BBI已被证明具有抗炎作用，而没有相关毒性。尽管BBI已经进行了临床测试，但其抗炎作用的机制还知之甚少。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免疫细胞的影响。在我们之前的研究中，我们发现BBI增加了白介素10(IL-10)的产生。IL-10具有众所周知的抗炎特性，我们将研究该分子是否在BBI的有益作用中发挥作用。已公布的数据和初步研究表明，BBI对两种关键的免疫细胞--即所谓的抗原提呈细胞和T细胞--有影响。该提案的第二部分致力于对BBI对这些细胞的影响进行机械性研究。我们相信，这些研究将为BBI的抗炎作用提供深入的见解，并有助于将其用作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.