DEFENSE MECHANISMS OF LUNG CELLS AGAINST MYCOBACTERIUM TUBERCULOSIS

这份续期申请的重点是剖析辩方 正常肺组织抗典型分枝杆菌的机制 结核病（MTB），并了解这些机制如何 可能在HIV感染中受损。我们将从我们的 外周血来源的巨噬细胞（PBM）和 鸟分枝杆菌的新实验，使用初级 支气管肺泡灌洗（BAL）细胞和MTB。这些实验将 比较正常BAL细胞与感染了 体外HIV-1和来自HIV感染者的BAL细胞 个体我们打算把正常的辩护 这些细胞对MTB的机制，然后学习，如果具体 缺陷出现在体外和体内的HIV环境中。我们 初步数据和其他人最近的工作表明， 细胞因子IL-2、IL-7、IL-10、IL-12、IL-13、TNF-α、IFN-γ和 纤溶酶原激活物抑制剂2型（派-2）参与 巨噬细胞和淋巴细胞功能的调节 分枝杆菌感染的背景。我们会研究 MTB攻击后BAL细胞中这些细胞因子的诱导 以及在测定中BAL细胞对激活细胞因子的反应 山地车杀人我们还将研究派-2的作用（我们 先前已显示可挽救感染分枝杆菌的PBM 在肺泡巨噬细胞（AM）实验中， 感染HIV-1或AM的HIV+患者。这些研究 旨在澄清我们假设的有效性， HIV感染者肺防御功能受损 针对MTB的缺陷是由于以下一个或多个缺陷： 1. T细胞功能障碍导致细胞因子受损 产生，例如不能产生IL-2、IL-7和IFN-γ，或 IL-10或IL-13的过度产生，2. AM的失败 在分枝杆菌感染中有效活化，例如由于 功能失调的细胞因子效应或HIV的直接效应，和3. AM无法在分枝杆菌感染中存活，可能是由于 派-2的保护功能障碍。我们的目标是 确定免疫治疗的候选者，以补充可用的抗- 结核病化疗可能对艾滋病的疗效有限 患者为此，我们还将研究 抗分枝杆菌反应可通过添加 脂阿拉伯甘露聚糖从无毒MTB，我们将探索 使用分枝杆菌噬菌体靶向治疗 分子治疗和MTB的直接裂解。

This renewal application focuses on the dissection of the defense mechanisms of the normal lung against typical Mycobacterium tuberculosis (MTB) and on the understanding how these mechanisms may be impaired in HIV infection. We will carry insights from our previous work on peripheral blood derived macrophages (PBM) and Mycobacterium avium to new experiments using primary bronchoalveolar lavage (BAL) cells and MTB. These experiments will compare functions of normal BAL cells to BAL cells infected with HIV-1 in vitro and with BAL cells derived from HIV-infected individuals. We intend to characterize the normal defense mechanisms of these cells against MTB, then to learn if specific defects arise in the setting of HIV in vitro and in vivo. Our preliminary data, and recent work by others, suggests that the cytokines IL-2, IL-7, IL-1O, IL-12, IL-13, TNF-alpha, IFN-gamma and the plasminogen activator inhibitor type 2 (PAI-2) participate in the modulation of the functions of macrophages and lymphocytes in the context of a mycobacterial infection. We will examine the induction of these cytokines in BAL cells following MTB challenge and the response of BAL cells to activating cytokines in assays of MTB killing. We will also examine the actions of PAI-2 (which we have previously shown to rescue PBM infected with Mycobacterium avium from death) in experiments with alveolar macrophages (AM) infected with HIV-1 or AM from HIV+ patients. These studies are designed to clarify the validity of our hypothesis that the impaired pulmonary defense function of HIV-infected individuals against MTB is due to one or more of the following defects: 1. T cell dysfunction resulting in impairment of cytokine production, e.g. failure to produce IL-2, IL-7 and IFN-gamma or hyperproduction of IL-10 or IL-13, 2. failure of the AM to be effectively activated in mycobacterial infection, e.g. due to dysfunctional cytokine effects or direct effects of HIV, and 3. incapacity of AM to survive mycobacterial infection possibly due to dysfunction of the protective function of PAI-2. Our goal is to identify candidates for immunotherapy to supplement available anti- tuberculous chemotherapy which may be of limited benefit in AIDS patients. To this end we will also examine whether the antimycobacterial response can be augmented by addition of lipoarabinomannans from nonvirulent MTB and we will explore therapeutic strategies employing mycobacteriophages for targeted molecular therapy and for direct lysis of MTB.