STRATEGIES FOR TUBERCULOSIS VACCINE DEVELOPMENT AND SCRE

本申请旨在响应PA-97-101“疫苗有效性的基本机制”，使用高度相关和相关的结核病感染模型，对疫苗诱导的肺部免疫机制进行基础研究。 一个潜在的动机是目前在小鼠和豚鼠模型中筛选的大量新的结核病候选疫苗，其中科罗拉多州立大学的分枝杆菌研究实验室（MRL）正在发挥核心作用。 随着测试的候选疫苗的数量逐渐增加，越来越明显的是，关于针对这种严重的细菌性呼吸道病原体的疫苗策略的基本信息，特别是关于肺中免疫力的表达，几乎完全缺乏。 因此，本申请提出解决四个主要关注领域，在每种情况下都有可能获得重要的新信息，这些信息可能会改善和扩大我们对结核病疫苗接种的整体方法。 这些领域是[1]开发一种广泛适用的方法来鉴定潜在的蛋白质抗原，并将这种方法具体应用于鉴定结核分枝杆菌的关键靶抗原，这种方法将借鉴我们在细菌培养、抗原提取和蛋白质组学分析方面的大量经验;[2]基于我们最近在这一领域的成功，进一步探索疫苗接种的细胞因子增强潜力; [3]努力确定疫苗接种下调的基础，特别是在干扰BCG疫苗方面;和[4]肺中CD 8 T细胞应答的性质和抗原靶点以及CD 8定向疫苗接种策略的潜力。 这些研究将利用我们广泛的最先进的III级生物危害设施，我们在批量培养和抗原纯化方面的专业知识，我们新获得的复杂细胞分选技术，以及我们在使用和开发小鼠低剂量气溶胶结核病感染模型方面的经验。

The purpose of this application in response to PA-97-101, "Basic mechanisms of vaccine efficacy", is to conduct basis research into mechanisms of vaccine-induced immunity in the lungs, using a highly pertinent and relevant tuberculosis infection model. An underlying motive is the plethora of new TB vaccine candidates currently being screened in mouse and guinea pig models, in which the Mycobacteria Research Laboratories [MRL] at Colorado State University are playing a central role. As the number of vaccine candidates tested has gradually increased, it has become increasingly apparent that fundamental information regarding vaccine strategies against this serious bacterial respiratory pathogen, particularly as it pertains to expression of immunity in the lungs, is almost completely lacking. Accordingly, this application proposes to address four major areas of concern, which in each case has the potential to derive important new information which may improve and broaden our overall approach to tuberculosis vaccination. These areas are [1] development of a widely applicable approach to identification of potential protein antigens, and specific application of this approach to the identification of key target antigens of Mycobacterium tuberculosis, an approach that will draw on our considerable experience in bacterial cultivation, antigen extraction, and proteomic analysis; [2] a further exploration of the potential for cytokine enhancement of vaccination, based on our recent success in this area; [3] work to determine the basis of down-regulation in vaccination, specifically in terms of interference with the BCG vaccine; and [4] the nature and antigenic targets of the CD8 T cell response in the lungs and the potential for CD8-directed vaccination strategies. These studies will exploit our extensive state of the art Level III biohazard facilities, our expertise in bulk cultivation and antigen purification, our new access to sophisticated cell sorting technology, and our demonstrated experience in using and developing the mouse low dose aerosol tuberculosis infection model.