Jimenez Victor, Monroy Fernando P
Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ, USA.
Bio Protoc. 2019 Jan 20;9(2):e3143. doi: 10.21769/BioProtoc.3143.
Alcohol consumption has diverse and well-documented effects on the human immune system and its ability to defend against infective agents. While pulmonary related infections can occur in healthy humans, binge alcohol use is recognized as a major health risk factor (Nelson , 1991). Although binge alcohol consumption has been considered as a risk factor for the development of pulmonary infections, no experimental studies have investigated the outcomes of a single binge alcohol exposure during infection. A key assay to assess the effects of a single binge alcohol exposure on the interactions between bacteria and alveolar macrophage is a binge alcohol intracellular invasion and killing assay. MH-S alveolar macrophages (AMs) are exposed to a single binge alcohol dose prior to infection for 3 h. The macrophage monolayer is then infected to allow for engulfment, followed by removal of extracellular bacteria to assess the intracellular killing capacity of infected macrophages over time. We have utilized this assay to demonstrate that low alcohol exposure significantly suppressed the uptake and killing of less virulent ) by AMs. More recently we found that activated AMs with interferon (IFN)-γ incubated in alcohol (0.08%) for 3 h prior to infection showed significantly lower bacterial uptake at 2 and 8 h post infection, which lead to survival and a ~2.5-fold replication increase compared to controls (Jimenez , 2017). These results provide insights into binge alcohol consumption, a culturally prevalent risk factor, as a predisposing factor for pulmonary bacterial infections. This assay can be adapted to other bacterial species and host cell types to assess tissue specific effects of alcohol during infection.
饮酒对人体免疫系统及其抵御感染因子的能力具有多种且有充分文献记载的影响。虽然健康人也可能发生肺部相关感染，但大量饮酒被认为是一个主要的健康风险因素（纳尔逊，1991年）。尽管大量饮酒被视为肺部感染发生的一个风险因素，但尚无实验研究调查感染期间单次大量饮酒的后果。评估单次大量饮酒对细菌与肺泡巨噬细胞相互作用影响的一个关键检测方法是大量饮酒细胞内侵袭和杀伤检测。在感染前3小时，将MH-S肺泡巨噬细胞（AMs）暴露于单次大量饮酒剂量。然后感染巨噬细胞单层以使其吞噬，随后去除细胞外细菌，以评估感染巨噬细胞随时间的细胞内杀伤能力。我们利用该检测方法证明，低剂量酒精暴露显著抑制了AMs对毒性较低细菌的摄取和杀伤。最近我们发现，在感染前用干扰素（IFN）-γ激活的AMs在酒精（0.08%）中孵育3小时，在感染后2小时和8小时细菌摄取显著降低，这导致细菌存活且与对照组相比复制增加约2.5倍（希门尼斯，2017年）。这些结果为大量饮酒这一文化上普遍存在的风险因素作为肺部细菌感染的易感因素提供了见解。该检测方法可适用于其他细菌种类和宿主细胞类型，以评估感染期间酒精对组织的特异性影响。
