HOST RESPONSE TO TB AND AIDS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。美国最近的结核病流行是由几个因素造成的，包括公共卫生项目资金不足、城市无家可归者收容所和监狱人满为患、结核病高发国家不断向美国移民以及艾滋病毒流行。后一个因素可能是最重要的，特别是在艾滋病毒在注射吸毒者中最常见的地区，如纽约市的情况，至少33%的结核病病例发生在艾滋病毒感染者中。在较年轻的患者中，结核病和艾滋病毒的合并感染更为普遍：纽约市25-44岁年龄组的结核病患者中有整整60%同时存在艾滋病毒感染。艾滋病毒感染患者对结核病感染和疾病的易感性增加与宿主免疫力受损直接相关。近年来，我们和其他人已经阐明了宿主对结核病反应的关键成分，现在似乎越来越清楚的是，th1型t淋巴细胞反应与结核病患者的良好预后相关。此外，越来越多的证据表明，感染艾滋病毒的患者Th1数量和功能受损，导致干扰素- γ (IFN-g)缺乏，干扰素- γ是结核病宿主免疫的关键效应细胞因子。我们假设雾化的IFN-g将促进干扰素反应因子介导的Th1反应，从而减少HIV-1复制和良好的临床结果。为了验证这一假设，我们建议使用强大的研究工具支气管肺泡灌洗（BAL）对结核患者肺段的炎症环境进行取样，并将结果与同一患者和正常对照的未受损伤肺段进行比较。特异性目标1将比较30例HIV-1/TB合并感染患者的bal前和bal后标本，其中一半随机接受雾化IFN-g，终点是Th1反应和HIV-1病毒载量的测量。Specific Aim 2将研究IFN-g促进宿主防御的机制，包括共刺激分子、MHC II类和诱导型一氧化氮合酶的研究。Specific Aim 3将由Richard Pine博士（公共卫生研究所）分析IFN-g信号传导的分子机制，包括STAT分子、它们的磷酸化、IFN-g调节因子1和II类反激活子。这些研究的结果将进一步表征体内局部宿主对结核分枝杆菌的反应，并确定宿主反应是否可以被IFN-g调节，特别是在HIV-1/TB合并感染的患者中，从而产生更有利的临床结果。该实验室用于以下工作：BAL、DNA分离、DNA测序（自动化）、ELISA、寡核苷酸合成、PCR、重组DNA技术、RNA分离、Northern分析、血液分离、Western分析、EMSA和层流罩的使用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The recent tuberculosis (TB) epidemic in the United States has been fueled by several factors, including underfunded public health programs, overcrowding in urban homeless shelters and prisons, continuing immigration to the United States from countries with a high incidence of TB, and the HIV epidemic. This latter factor may be the most significant, particularly in areas where HIV is most common among injection drug users, as is the case in New York City, where at least 33% of TB cases occur in HIV-infected persons. In younger patients, co-infection between TB and HIV is even more prevalent: fully 60% of TB patients in the 25-44 year-old age group in New York City have co-existing HIV infection. The increased susceptibility to TB infection and disease among HIV-infected patients is directly related to impaired host immunity. In recent years, we and others have elucidated key components of the host response to TB, and it now seems increasingly clear that a Th1-type T-lymphocyte response is associated with a good outcome in TB patients. In addition, there is increasing evidence that HIV-infected patients have impaired Th1 number and function, with a resultant deficiency of interferon-gamma (IFN-g), a key effector cytokine in host immunity in TB. We hypothesize that aerosolized IFN-g will promote a Th1 response mediated by interferon-responsive factors leading to reduced HIV-1 replication and a propitious clinical outcome. To test this hypothesis we propose to use the powerful research tool of bronchoalveolar lavage (BAL) to sample the inflammatory milieu of lung segments with TB and compare results to uninvolved segments of the same patient and normal controls. Specific Aim 1 will compare pre- to post-BAL specimens in 30 HIV-1/TB co-infected patients, with half randomized to receive aerosolized IFN-g and the endpoints being measurements of Th1 response and HIV-1 viral load. Specific Aim 2 will investigate mechanisms by which IFN-g contributes to host defense, including studies of co-stimulatory molecules, MHC class II and inducible nitric oxide synthase. Specific Aim 3 will analyze molecular mechanisms of IFN-g signalling by Richard Pine, Ph.D. (Public Health Research Institute), including STAT molecules, their phosphorylation, IFN-g regulatory factor 1, and class II transactivator. Findings from these studies will further characterize the local host response to M. tuberculosis in vivo and determine if the host response can be modulated, particularly in HIV-1/TB co-infected patients, by IFN-g, thus resulting in a more favorable clinical outcome. The lab was utilized for the following: BAL, DNA isolation, DNA sequencing (automated), ELISA, oligonucleotide synthsis, PCR, recombinant DNA techniques, RNA isolation, Northern analysis, blood separation, Western analysis, EMSA, and use of laminar flow hoods.