BACTERIAL GROWTH IN MACROPHAGES FROM ALCOHOL FEEEDING

描述：过度饮酒使个人处于更高的水平 严重细菌感染的风险，包括细胞内感染的风险 病原体。这个实验室和其他实验室的观察表明 巨噬细胞功能的许多方面，包括细胞因子的表达和 抗原提呈，在使用酒精的个体中受到损害。 因此，这项提案旨在确定巨噬细胞是否以及如何 从酒精喂养的小鼠身上分离出来的细菌更容易产生细菌 感染。最有效地探测酒精诱导的巨噬细胞 为了改变，我们在我们的实验设计中加入了六个功能。 首先，我们将检查以下三种病原体的感染情况 细胞内途径，即单核细胞增多性李斯特氏菌、沙门氏菌 鼠伤寒杆菌和嗜肺军团菌。第二，我们将评估增长 在三个品系的小鼠的巨噬细胞内表现出不同的 对这些细菌的敏感程度。第三，实验只会 利用病原体在感染过程中遇到的巨噬细胞 正常感染；例如，李斯特菌和肺泡炎的脾细胞 军团菌的巨噬细胞。第四，更充分地阐明 在感染过程中，我们将在多个时间点监测人数 细胞内细菌的数量，宿主细胞的活性程度，以及 IL-1、IL-6、IL-12和肿瘤坏死因子的产生。第五，认识到 酒精对先天免疫和获得性免疫的影响，包括常驻和激活的 巨噬细胞将接受敏感性变化测试。最后，我们 将决定是否添加T细胞或促炎细胞因子， 包括干扰素-γ在内，可以“逆转”乙醇的作用。总体而言， 此R21应用程序中提出的实验应该提供新的洞察力 酒精性感染性疾病的分子和细胞基础 为其他独特的调查路线奠定基础。

DESCRIPTION: Excessive alcohol consumption places individuals at higher risk for serious bacterial infections, including those by intracellular pathogens. Observations by this laboratory as well as others indicate that many aspects of macrophage function, including cytokine expression and antigen presentation, are compromised in ethanol consuming individuals. Consequently, this proposal aims to determine if and how macrophages explanted from alcohol-fed mice are more permissive for productive bacterial infection. To most effectively probe the macrophage for alcohol-induced changes, we have incorporated six features into our experimental design. First, we will examine infection by three pathogens which follow distinct intracellular pathways, namely Listeria monocytogenes, Salmonella typhimurium, and Legionella pneumophila. Second, we will assess growth within macrophages from three strains of mice which display different degrees of susceptibility to these bacteria. Third, experiments will only utilize those macrophages which the pathogens encounter in the course of normal infections; e.g., splenic cells for Listeria, and alveolar macrophages for Legionella. Fourth, to more fully illuminate changes in the infection process, we will monitor, at a multiple time points, the numbers of intracellular bacteria, the degree of host cell viability, and the levels of IL-1, IL-6, IL-12, and TNF produced. Fifth, to appreciate the effect of alcohol on innate as well as adaptive immunity, both resident and activated macrophages will be tested for alterations in susceptibility. Finally, we will determine whether the addition of T cells or proinflammatory cytokines, including IFN-gamma, can "reverse" the effect of ethanol. Collectively, the experiments proposed in this R21 application should provide new insight into the molecular and cellular bases of alcohol-induced infectious diseases and lay the foundation for additional, unique lines of inquiry.