Innate Immunity Driven Th1/Th2 bias in Leishmaniasis

描述(由申请者提供)：候选人Keith G.Nelson，DVM的直接目标是完成一项关于利什曼原虫感染免疫病理研究的指导性培训计划，最终将获得博士学位和继续博士后研究。纳尔逊博士的长期目标是成为一名首席研究员，研究利什曼病和其他媒介传播的寄生性病原体的免疫反应、发病机制和预防。研究培训计划将以科罗拉多州立大学免疫寄生虫学研究组理查德·G·蒂图斯博士的实验室为中心。研究环境有着强大的外部资金和生物医学科学家培训的历史。候选人的培训计划包括应用于利什曼病动物模型的现代分子和免疫学技术和研究方法的实验室和教学指导。 这项研究利用已建立的利什曼原虫感染小鼠模型来研究大利什曼原虫(LM)免疫应答的早期阶段以及随后的1型T细胞与2型T细胞(Th1/Th2)的分化，重点关注参与宿主识别和反应的巨噬细胞受体和寄生虫表面抗原。将致力于三个具体目标：1)识别参与识别、吞噬和应答LM的巨噬细胞补体和非补体受体，以及表征通过特定巨噬细胞受体进入宿主后Th1/Th2应答的变化；2)表征Toll样受体(Toll样受体)在转导宿主对LM感染的应答中的作用，识别相关受体并确定对Th1/Th2偏向的影响；3)确定主要寄生虫表面分子脂磷脂多糖(LPG)和gp63对宿主识别、吞噬和Th1/Th2应答的影响。所有这些都将通过在体内感染相关的LPG或gp63缺陷寄生虫敲除小鼠，以及通过体外巨噬细胞培养、受体阻断、感染和与幼稚Th细胞共培养来评估功能启动。细胞因子谱将通过酶联免疫吸附试验和逆转录聚合酶链式反应进行评估，以确定Th1/Th2相关偏倚，TLR mRNA将通过逆转录聚合酶链式反应进行评估。 这些研究的结果将为研究先天免疫反应的初级吞噬细胞与原虫寄生虫利什曼原虫之间的相互作用提供新的见解。我们还将深入了解先天免疫反应和病原体识别途径在适应性免疫中Th1/Th2反应建立中的作用。这些信息可能被证明对开发有效的治疗、控制和预防措施以及进一步了解免疫系统的先天和适应性手臂之间的相互作用具有重要意义。

DESCRIPTION (provided by applicant): The immediate goal of the candidate, Keith G. Nelson, DVM, is to complete a mentored training program in research of immunopathogenesis of Leishmania infection, which will culminate in both the PhD degree and continued postdoctoral study. Dr. Nelson's long-term goal is to become a principal investigator conducting research on the immune response to, pathogenesis and prevention of leishmaniasis and other vector-borne parasitic pathogens. The research-training program will be centered on the laboratory of Dr. Richard G. Titus, in the immunoparasitology Research Group at Colorado State University. The research environment has a strong history of extramural funding and biomedical scientist training. The candidate's training plan consists of laboratory and didactic instruction in contemporary molecular and immunological techniques and research methods applied to an animal model of leishmaniasis. The proposed research utilizes an established murine model of Leishmania infection to investigate the early stages of the immune response to Leishmania major(Lm) and subsequent Type 1 T cell vs. Type 2 T cell (Th1/Th2) differentiation, with a focus on the macrophage receptors and parasite surface antigens involved in host recognition and response. Three specific aims will be addressed: 1) to identify the macrophage complement and non-complement receptors involved in recognition, phagocytosis, and response to Lm, as well as characterizing changes in the Th1/Th2 response secondary to entry via specific macrophage receptors; 2) to characterize the role of toll-like receptors TLRs) in transducing host response to Lm infection, identifying relevant receptors and determining the effect on Th1/Th2 bias; 3) to determine the effect of major parasite surface molecules lipophosphoglycan (LPG) and gp63 on host recognition, phagocytosis and Th1/Th2 response to Lm. All of these will be addressed using in vivo infection of relevant knockout mice with LPG or gp63 deficient parasites, as well as with in vitro macrophage culture, receptor blocking, infection and co-cultivation with naive Th cells to assess functional priming. Cytokine profiles will be assessed via ELISA and RTPCR to determine Th1/Th2-associated bias and TLR mRNA will be assessed via RT-PCR. The results of these studies will provide new insights into the interactions between the primary phagocytic ceils of the innate immune response and the protozoan parasite Leishmania major. We will also gain insight into the effect of the innate immune response and pathogen recognition pathway on the establishment of Th1/Th2 responses in adaptive immunity. This information may prove to be important in the development of effective treatment, control and preventative measures in leishmaniasis, as well as furthering our knowledge of the interactions between the innate and adaptive arms of the immune system.