Bovine epithelial and endothelial response to Mannheimia

描述（由申请人提供）：这个拟议的项目包含了一个阶段性的职业发展计划的候选人在传染病研究的长期利益。根据这一阶段性计划，前2年将包括方法开发，初步实验和专业发展，以进一步提高候选人在兽医和研究方面的背景。接下来的3年将用于开发方法，以深入研究肺内皮细胞和上皮细胞之间的相互作用，以应对牛呼吸道病原体溶血性曼氏菌。与人类呼吸系统疾病急性呼吸窘迫综合征（ARDS）一样，肺炎性巴氏杆菌病与血管成分严重渗漏到空气中有关。在感染的初始阶段，血管渗漏与肺中的中性粒细胞活化相关，而唇多糖可能是感染后期血管渗漏的介体。中心假设是M.溶血和嗜中性粒细胞活化都参与促进血浆渗漏。具体目的包括1）确定LPS和中性粒细胞活化对由肺泡上皮细胞和肺微血管内皮细胞组成的共培养系统中白蛋白渗漏和凋亡的影响，2）确定肺上皮细胞活化对肺微血管内皮细胞细胞因子产生的影响，3）观察LPS和细胞因子刺激后肺泡上皮细胞Toll样受体4（TLR4）的表达。了解中性粒细胞，LPS，上皮细胞和内皮细胞之间的相互作用，在肺炎巴氏杆菌病将导致更好地了解肺血管渗漏的基本机制。完成该项目后，候选人将获得新的实验室技术，培养批判性思维能力，并作为一名科学家专业成长，从而使候选人成为一名独立的调查员。

DESCRIPTION (provided by applicant): This proposed project incorporates a phased career development plan for a candidate with a long-standing interest in infectious disease research. Under this phased plan, the first 2 years will incorporate methods development, preliminary experimentation, and professional development to further the candidate's background in veterinary medicine and research. The following 3 years will be spent developing methods to intensively investigate interactions between lung endothelial and epithelial cells in response to the bovine respiratory agent Mannheimia haemolytica. Like the human respiratory condition acute respiratory distress syndrome (ARDS), pneumonic pasteurellosis is associated with profound leakage of vascular components into the airspaces. Vascular leakage is associated with neutrophil activation in the lung during the initial stage of infection, whereas lip polysaccharide is likely a mediator of vascular leakage later in infection. The central hypothesis is that LPS from M. haemolytica and neutrophil activation are both involved in promoting plasma leakage. The specific aims include 1) determine the effect of LPS and neutrophil activation on albumin leakage and apoptosis in a co-culture system consisting of alveolar epithelial cells and pulmonary micro vascular endothelial cells, 2) determine the effect of lung epithelial cell activation on pulmonary micro vascular endothelial cell cytokine production, and 3) characterize the expression of Toll-like receptor 4 (TLR4) on alveolar epithelial cells after LPS and cytokine stimulation. Understanding the interactions between neutrophils, LPS, epithelial cells, and endothelial cells during pneumonic pasteurellosis will result in a better understanding of the basic mechanisms of vascular leakage in the lung. Upon completion of this project, the candidate will have acquired new laboratory techniques, developed critical thinking skills, and grown professionally as a scientist, leading to the candidate's progression into an independent investigator.