"The Tuberculous Granuloma"

这一重返申请的主要目标是促进帕特里克比尔德博士的重新参与， 科学比尔德博士最近经历了两年半的职业生涯转移 因病重返大气层申请的科学部分，其目的是确定 一组结核分枝杆菌（Mtb）突变体的细胞包膜成分， 增强巨噬细胞肿瘤坏死因子（TNF）产生的能力，来自结果 由正在进行的P01项目生成，我是项目3的PI。一个主要组成部分 P01计划项目3的目标是通过靶向Mtb TNF调节来开发有效的结核病疫苗， 基因以产生具有增强的免疫原性的突变体。该策略基于 TNF促进细胞凋亡和吞噬溶酶体融合（两种细胞过程）能力 已知通过增强免疫反应的交叉致敏来增强免疫反应的免疫原性， 细胞毒性CD 8 T细胞和抗原呈递。通过基因筛选，我们得到了 鉴定了一组Mtb基因，其破坏导致具有TNF上调的突变体， 感染巨噬细胞中的表型。根据TNF和TNF-α的上述性质， 验证我们的疫苗开发策略，这些突变体促进凋亡， 巨噬细胞吞噬溶酶体融合及C57 BL/6小鼠免疫试验 所引发Th 1应答上级野生型（WT）Mtb菌株H37 Rv所引发的应答。这些 TNF-上调突变体被破坏的基因已被注释为编码 能够调节Mtb细胞包膜成分的功能。这个项目的总体目标是 应用是检验TNF上调Mtb突变体的细胞包膜 与WT菌株不同，因此导致与宿主的相互作用改变 感染后，巨噬细胞的TNF产生增加。到 为了确保比尔德博士重返科学界的成功，我们制定了一个计划， 包括板凳经验，职业发展机会，多个场地的科学 互动，和教学课程;此外，一个完善的结核病研究人员， 与提案主题相关的经验--Jordi Torrelles博士（俄亥俄州 大学）已被列为博士比尔德的共同导师。我们相信这项提案将 成功地促进比尔德博士重新进入科学界，并可能产生的信息， 阐明了Mtb缺失增强TNF属性的机制 菌株，这反过来可能导致开发新的有效的抗结核疫苗。

The major goal of this reentry application is to facilitate Dr. Patrick Bilder’s reengagement with science. Dr. Bilder has experienced a recent two-and-a-half year diversion from his career path due to illness. The scientific component of the re-entry application, which aims at characterizing the cell envelope constituents of a set of Mycobacterium tuberculosis (Mtb) mutants with the ability to enhance macrophage tumor necrosis factor (TNF) production, derives from the results generated by an on-going P01 program, on which I am the PI of Project 3. A major component of Project 3 of the P01 Program is to develop effective TB vaccines by targeting Mtb TNFregulating genes to generate mutants with enhanced immunogenecity. This strategy is based on the ability of TNF to promote apoptosis and phagolysosomal fusion, two cellular processes known to augment immunogenicity of an immunological reaction by enhancing cross-priming of cytotoxic CD8 T cells and antigen presentation; respectively. Using a genetic screen, we have identified a set of Mtb genes whose disruption results in mutants with TNF-upregulating phenotype in infected macrophages. In line with the above-stated properties of TNF and validating our vaccine development strategy, these mutants promote apoptosis and phagolysosomal fusion in macrophages and imunization of C57BL/6 mice with these strains elicit a Th1 response superior to that engendered by the wildtype (WT) Mtb strain H37Rv. These TNF-upregulating mutants are disrupted for genes that have been annotated to encode functions capable of modulating the Mtb cell envelope components. The overall goal of this application is to test the hypothesis that the cell envelope of the TNF-upregulating Mtb mutants are different than that of the WT strain, thus resulting in altered interaction with host macrophages, leading to enhanced TNF production by the phagocytes upon infection. To ensure the success of Dr. Bilder’s re-entry into science, we have in place a plan that incorporates bench experience, career development opportunities, multiple venues for scientific intearctions, and didactic courses; in addition, a well established TB investigators with experience pertinent to the subject matter of the proposal -- Dr. Jordi Torrelles (Ohio State University) has been enlisted as Dr. Bilder’s Co-mentor. We believe this proposal will successfully facilitate Dr. Bilder’s re-entry into science and will likely yield information that will shed light on the mechanisms underlying the TNF-enhancing attributes of the Mtb deletion strains, which may, in turn, lead to the development of novel effective anti-TB vaccines.