Sharma Lokesh, Wu Wenjun, Dholakiya Sanjay L, Gorasiya Samir, Wu Jiao, Sitapara Ravikumar, Patel Vivek, Wang Mao, Zur Michelle, Reddy Shloka, Siegelaub Nathan, Bamba Katrina, Barile Frank A, Mantell Lin L
Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, 128 St. Albert Hall, 8000 Utopia Parkway, 11439, Queens, NY, USA.
Methods Mol Biol. 2014;1172:137-45. doi: 10.1007/978-1-4939-0928-5_12.
Phagocytosis is the process by which phagocytes, including macrophages, neutrophils and monocytes, engulf and kill invading pathogens, remove foreign particles, and clear cell debris. Phagocytes and their ability to phagocytose are an important part of the innate immune system and are critical for homeostasis of the host. Impairment in phagocytosis has been associated with numerous diseases and disorders. Different cytokines have been shown to affect the phagocytic process. Cytokines including TNFα, IL-1β, GM-CSF, and TGF-β1 were found to promote phagocytosis, whereas high mobility group box-1 (HMGB1) inhibited the phagocytic function of macrophages. Here, we describe two commonly used methods to assess the phagocytic function of cultured macrophages, which can easily be applied to other phagocytes. Each method is based on the extent of engulfment of FITC-labeled latex minibeads by macrophages under different conditions. Phagocytic activity can be assessed either by counting individual cells using a fluorescence microscope or measuring fluorescence intensity using a flow cytometer.
吞噬作用是指包括巨噬细胞、中性粒细胞和单核细胞在内的吞噬细胞吞噬并杀死入侵病原体、清除外来颗粒和清除细胞碎片的过程。吞噬细胞及其吞噬能力是固有免疫系统的重要组成部分，对宿主的内环境稳态至关重要。吞噬作用受损与多种疾病和病症相关。已表明不同的细胞因子会影响吞噬过程。发现包括肿瘤坏死因子α、白细胞介素-1β、粒细胞-巨噬细胞集落刺激因子和转化生长因子-β1在内的细胞因子可促进吞噬作用，而高迁移率族蛋白B1（HMGB1）则抑制巨噬细胞的吞噬功能。在此，我们描述两种常用的评估培养巨噬细胞吞噬功能的方法，这些方法可轻松应用于其他吞噬细胞。每种方法都基于在不同条件下巨噬细胞对异硫氰酸荧光素标记的乳胶微珠的吞噬程度。吞噬活性可通过使用荧光显微镜计数单个细胞或使用流式细胞仪测量荧光强度来评估。
