Regulate/Lymphocyte Traffic/Chemokines/Adhesion/Crohn's

描述（由申请人提供）： TNF阻断治疗克罗恩病（CD）的成功导致了对其他生物疗法的系统研究。不幸的是，这些新疗法（如IL-10、IL-11阻断）的有效性受到限制。因此，必须在CD中评价靶向其他慢性肠道炎症途径的替代生物疗法。我们建议，由于冗余的淋巴细胞粘附级联反应，同时封锁两个或两个以上的肠道归巢粘附分子和/或趋化因子（即L-选择素，β 7整合素，MAdCAM-1和CCR 9）将需要。因此，我们的中心假设是，通过靶向粘附分子/趋化因子的特定组合来干扰淋巴细胞运输将导致CD的TNFdeltaARE小鼠模型中慢性回肠炎的改善。 为了解决这一假设，我们提出以下具体目标： 1.确定慢性小肠炎症条件下淋巴细胞粘附级联反应的冗余。使用免疫组化，PCR，流式细胞术和活体显微镜，我们将解剖淋巴细胞运输的途径，在慢性回肠炎，并确定支持淋巴细胞粘附和迁移的分子。 2.评估肠道归巢粘附分子/趋化因子在慢性回肠炎发生中的作用。我们将TNFdeltaARE小鼠和TNFdeltaARE/β 7-/-小鼠与缺乏L-选择素和CCR 9的小鼠回交。 3.评价粘附分子/趋化因子联合阻断治疗慢性回肠炎的有效性。为了实现这一转化目标，我们将使用特异性功能阻断单克隆抗体的组合，这些单克隆抗体可能会改善目前已在临床试验中的抗α 4或α 4 β 7整联蛋白阻断策略（即Natalizumab，MLN 02）的有效性。 本提案旨在为主要研究者提供各种实验室技术方面的丰富经验，如具体目标所列。2导师的优势，专门从事粘膜免疫学（FC）和粘附分子（KFL），以及实验所需的所有设备，试剂和小鼠品系的可用性是我们机构所独有的。这些经验将通过实验室和课堂教学的严格计划得到补充。鉴于人类CD和我们小鼠模型中TNF诱导的慢性回肠炎之间的相似性，拟议的研究也可以为治疗这种毁灭性疾病的新生物靶点提供重要线索。

DESCRIPTION (provided by applicant): The success of TNF blockade for the treatment of Crohn's disease (CD) has lead to the systematic study of other biological therapies. Unfortunately, the usefulness of these newer therapies (e.g. IL-10, IL-11 blockade) has been limited. Therefore, alternative biological therapies that target other pathways of chronic intestinal inflammation must be evaluated in CD. We propose that due to redundancies in the lymphocyte adhesion cascade, simultaneous blockade of two or more gut-homing adhesion molecules and/or chemokines (i.e. L-selectin, beta7integrin, MAdCAM-1 and CCR9) will be required. Therefore, our central hypothesis is that interference with lymphocyte trafficking by targeting specific combinations of adhesion molecules/chemokines will result in amelioration of chronic ileitis in the TNFdeltaARE murine model of CD. To address this hypothesis we propose the following specific aims: 1. To identify redundancies of the lymphocyte adhesion cascade under conditions of chronic small intestinal inflammation. Using immunohistochemistry, PCR, flow cytometry and intravital microscopy we will dissect the pathways of lymphocyte trafficking in chronic ileitis and identify the molecules that support lymphocyte adhesion and migration. 2. To assess the role of gut-homing adhesion molecule/chemokines on the development of chronic ileitis. We will backcross TNFdeltaARE mice and TNFdeltaARE/beta7-/- mice to mice deficient for L-selectin and CCR9. 3. To evaluate the effectiveness of combinatorial blockade of adhesion molecules/chemokines for the treatment of established chronic ileitis. For this translational aim, we will use combinations of specific function-blocking monoclonal antibodies that may improve the effectiveness of current anti-alpha4- or alpha4beta7- integrin blockade strategies (i.e. Natalizumab, MLN02), already in clinical trials. This proposal is designed to provide the principal investigator with extensive experience in a variety of laboratory techniques, as listed on the specific aims. The strengths of 2 mentors that specialize in mucosal immunology (FC) and adhesion molecules (KFL), as well as the availability of all equipment, reagents and mouse strains required for the experiments is unique to our institution. These experiences will be complemented by a rigorous program of laboratory and classroom instruction. Given the similarities between human CD and the TNF-induced chronic ileitis in our mouse model, the proposed studies could also provide important leads for novel biological targets to treat this devastating disease.