CLINICAL TRIAL: HOST RESPONSE TO TB AND AIDS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 最近美国结核病(TB)的流行是由几个因素助长的，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，继续从结核病高发国家移民到美国，以及艾滋病毒流行。后一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，那里至少33%的结核病病例发生在艾滋病毒感染者中。在年轻患者中，结核病和艾滋病毒之间的混合感染甚至更为普遍：纽约市25-44岁年龄组中整整60%的结核病患者同时存在艾滋病毒感染。艾滋病毒感染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，我们和其他人阐明了宿主对结核病的反应的关键成分，现在似乎越来越清楚地表明，Th1型T淋巴细胞反应与结核病患者的良好结局有关。此外，越来越多的证据表明，HIV感染患者Th1细胞数量和功能受损，导致干扰素-γ(IFN-g)缺乏，干扰素-γ是结核病宿主免疫中的关键效应细胞因子。我们假设雾化干扰素-g将促进由干扰素反应因子介导的Th1反应，从而减少HIV-1的复制并获得有利的临床结果。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)来采样肺结核患者肺段的炎性环境，并将结果与同一患者和正常对照组的正常肺段进行比较。特定目标1将比较30名HIV-1/TB混合感染患者BAL前后的样本，其中一半随机接受雾化干扰素-g治疗，终点是Th1应答和HIV-1病毒载量的测量。具体目标2将研究干扰素-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。具体目标3将分析Richard Pine博士(公共卫生研究所)传递干扰素-g信号的分子机制，包括STAT分子、它们的磷酸化、干扰素-g调节因子1和第二类反式激活因子。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干扰素-g调节，特别是在HIV-1/TB混合感染的患者中，从而产生更有利的临床结果。该实验室用于以下方面：BAL、DNA分离、DNA测序(自动)、ELISA、寡核苷酸合成、聚合酶链式反应、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.