Macrophage polarization and glutathione levels in the TB-helminth co-infection

描述（由申请方提供）：估计三分之一的人群潜伏感染M。结核病和每年约900万新病例的活动性结核病（TB）的报告。然而，只有大约30%的健康人密切接触结核病会受到感染，其中只有5%-10%会继续发展为活动性疾病-这表明在大多数情况下，有能力的免疫系统能够对抗感染。在低收入国家的贫困地区，结核病发病率特别高，在这些地区，蠕虫感染也很常见，有人提出，蠕虫感染引起的免疫学变化导致对结核病的易感性增加。人们普遍认为，细胞介导的免疫在控制初始TB感染中起关键作用，巨噬细胞是具有某种模糊的双重作用的关键参与者。巨噬细胞既可以作为一线效应细胞，也可以作为营养宿主细胞供真菌生长。对这种矛盾的部分解释可以在巨噬细胞的激活中找到。巨噬细胞可以是经典活化的（CAM）或替代活化的（AAM）。CAM是促炎性的，并保护宿主免受各种病原体的侵害，而AAM是抗炎性的，并且对于抑制炎症和愈合炎症引起的组织损伤很重要。如果在不适当的时间诱导一种巨噬细胞类型，可能会出现问题。据认为，蠕虫感染引起交替激活的巨噬细胞。我们认为，蠕虫诱导的巨噬细胞具有低水平的细胞内谷胱甘肽，这是一个因素，为增加结核病的易感性蠕虫感染的人。我们的目的是研究这种巨噬细胞极化和谷胱甘肽耗竭的存在以及对结核病巨噬细胞杀伤的影响。巨噬细胞将从有和没有蠕虫合并感染的TB患者、蠕虫感染者和健康对照者中分离。将使用luminex评估巨噬细胞活化特征，用于细胞因子;使用FACS评估AAM和CAM标志物，并使用比色测定法评估iNOS和agrinase活性。将使用荧光法测量谷氨酰胺水平。将使用荧光素酶标记的结核分枝杆菌菌株的感染试验评价谷胱甘肽耗竭对分枝杆菌杀灭的影响。

DESCRIPTION (provided by applicant): One third of the human population is estimated to be latently infected with M. tuberculosis and each year about 9 million new cases of active tuberculosis (TB) are reported. However, only about 30% of healthy people closely exposed to TB will get infected and of those only 5%-10% will go on to develop active disease - an indicative that a competent immune system is able to fight of the infection in the majority of cases. TB incidence is especially high in areas of poverty in low income countries, where infections with helminths also are common, and it has been proposed that immunological changes induced by a helminths infection leads to increased susceptibility to TB. It is generally accepted that cell mediated immunity play a pivotal role in controlling the initial TB infection, wth macrophages being key players with a somewhat ambiguous dual role. The macrophage can act both as a first-line effector cell, but also serve as a nutritious host cell for the mycobacterum to thrive in. Part of the explanation to this paradox may be found in the activation of the macrophage. A macrophage can be classically activated (CAM) or alternatively activated (AAM). The CAMs are pro-inflammatory and protects the host against various pathogens, whereas the AAMs are anti-inflammatory and are important for dampening inflammation and in the healing of tissue damage from inflammation. A problem may arise if one macrophage type is induced at an inappropriate time. It is thought that helminth infection give rise to alternatively activated macrophages. We suggest that helminths induced macrophages have low levels of intracellular glutathione and that this is a factor for the increased susceptibility to TB in helminths infected persons. We aim to investigate the presence of such a macrophage polarization and glutathione depletion and the effect on macrophage killing of TB. Macrophages will be isolated from patients with TB with and without helminths co-infection, helminths infected and healthy controls. Macrophage activation profile will be assessed using luminex for cytokines; FACS for AAM and CAM markers, and colorimetric assays for iNOS and agrinase activity. Glutathione levels will be measured using a fluorophoric method. The effect of glutathione depletion on mycobacterial killing will be evaluated using an infection assay with a luciferase tagged Mycobacterial tuberculosis strain.