Opiate Modulates Lymphocyte Trafficking into the CNS in TB Meningitis

描述（由申请人提供）：在全球范围内，结核病仍然是一项重大的公共卫生危机，艾滋病毒的流行大大加剧了这一危机。在临床上，结核分枝杆菌是艾滋病患者最重要的机会致病菌，而中枢神经系统感染是结核病最具破坏性的并发症。与艾滋病毒感染的情况一样，药物滥用被认为是结核分枝杆菌发病的潜在辅助因素。虽然已经有许多关于阿片类药物对其他细胞内微生物发病机制的影响的研究，但对阿片类药物对淋巴细胞运输的影响知之甚少，对阿片类药物对中枢神经系统防御结核分枝杆菌的影响一无所知。在中枢神经系统内，结核分枝杆菌引起神经炎症反应，但参与这种反应的细胞和介质在很大程度上是不明确的。虽然致敏的T淋巴细胞在防御许多中枢神经系统感染中起着关键作用，但T淋巴细胞进入大脑以应对感染因子（包括结核分枝杆菌）的过程尚未被描述。最近，一种利用转基因小鼠表达荧光素酶的新型成像系统被开发出来，使研究人员能够定量地追踪多发性硬化症动物模型中T淋巴细胞进入中枢神经系统的“体内”运输模式。本应用中提出的研究的总体目标是采用这种创新的方法来评估阿片类药物依赖对T淋巴细胞运输到中枢神经系统的影响，以应对脑内结核杆菌的挑战。这种定量成像系统的使用将使我们能够验证阿片类药物依赖会损害卡介苗致敏T淋巴细胞进入中枢神经系统，从而增加中枢神经系统结核病的严重程度的中心假设。为了验证这一假设，我们设计了实验，以解决以下两个特定目标：特定目标1：采用一种新的定量成像技术来确定卡介苗致敏的T淋巴细胞是否在对分枝杆菌感染的反应中进入中枢神经系统。据推测，卡介苗接种可通过在接种结核杆菌后卡介苗致敏的T淋巴细胞快速浸润到CNS中来保护小鼠免受CNS结核的侵袭。特异性目的2：确定吗啡是否增强了分枝杆菌感染的神经发病机制，并损害了T淋巴细胞进入中枢神经系统的转运。利用上述CNS结核模式和T细胞成像技术，吗啡给药对神经发病机制的影响将被阐明。我们推测吗啡依赖会加速死亡和增加感染引起的中枢神经系统损伤，并且吗啡依赖小鼠过继转移BCG致敏T淋巴细胞的运输会明显受损。

DESCRIPTION (provided by applicant): Globally, tuberculosis (TB) remains a major public health crisis, which has been dramatically fueled by the HIV epidemic. Clinically, Mycobacterium tuberculosis is the most important opportunistic pathogen in AIDS patients, and infection of the central nervous system (CNS) is the most devastating complication of TB. As in the case of HIV infection, drug abuse has been regarded as a potential cofactor in the pathogenesis of M. tuberculosis. While there have been numerous studies on the effects of opiates on the pathogenesis of other intracellular microbes, little is known about the influence of opiates on lymphocyte trafficking and nothing is known about the impact of opiates on defense of the CNS against M. tuberculosis. Within the CNS, M. tuberculosis elicits a neuroinflammatory response, but the cells and mediators involved in this response are largely undefined. Although sensitized T lymphocytes play a critical role in defense against many CNS infections, the process of T lymphocyte entry into the brain in response to infectious agents, including M. tuberculosis, has not been delineated. Recently, a novel imaging system utilizing transgenic mice expressing luciferase was developed allowing researchers to track quantitatively "in vivo" the trafficking pattern of T lymphocytes into the CNS in an animal model of multiple sclerosis. The overall goal of the research proposed in this application is to adapt this innovative methodology to assess the effects of opiate dependence on T lymphocyte trafficking into the CNS in response to intracerebral challenge with tubercle bacilli. The use of this quantitative imaging system will allow us to test the central hypothesis that opiate dependence impairs the trafficking of Bacille-Calmette-Gu¿rin (BCG)-sensitized T lymphocytes into the CNS thereby increasing the severity of CNS TB. To test this hypothesis, experiments have been designed which address the two specific aims outlined below: Specific aim 1: Adapt a novel quantitative imaging technique to determine whether BCG- sensitized T lymphocytes traffic into the CNS in response to mycobacterial infection. It is hypothesized that BCG vaccination will protect mice against CNS TB through a rapid infiltration of BCG- sensitized T lymphocytes into the CNS following ic inoculation of tubercle bacilli. Specific aim 2: Determine whether morphine administration enhances the neuropathogenesis of mycobacterial infection and impairs T lymphocyte trafficking into the CNS. Using the CNS TB paradigm above and T cell imagining technology, the effects of morphine administration on the neuropathogenesis will be elucidated. It is hypothesized that morphine dependence will accelerate mortality and increase CNS damage due to infection and that the trafficking of adoptive transferred BCG- sensitized T lymphocytes will be markedly impaired in morphine-dependent mice.