Channappanavar Rudragouda, Perlman Stanley
Department of Acute and Tertiary Care, University of Tennessee Health Science Center, Memphis, TN, USA.
Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, Memphis, TN, USA.
Methods Mol Biol. 2020;2099:195-204. doi: 10.1007/978-1-0716-0211-9_15.
Innate immune cells play a vital role in mounting an effective host response to a variety of pathogen challenges. Myeloid cells such as neutrophils and monocyte-macrophages are major innate leukocytes that orchestrate protective immunity to viral lung infections. However, a dysregulated cytokine response can promote excessive infiltration and robust pro-inflammatory activity of neutrophils and monocyte-macrophages, leading to fatal disease. Following virus infection, the beneficial or deleterious role of infiltrating neutrophils and monocyte-macrophages is determined largely by their ability to secrete inflammatory cytokines and chemokines. A majority of studies use the total number of infiltrating cells and their activation status as measures to demonstrate their role during an infection. Consequently, the ability of neutrophils and Inflammatory Monocyte Macrophages (IMMs) to secrete inflammatory cytokines and chemokines, and its correlation with the disease severity, is not well defined. In this chapter, we report useful markers to identify lung infiltrating innate immune cells and define their activation status. We also describe a simple method to measure intracellular cytokine production to evaluate the inflammatory activity of neutrophils and IMMs in a mouse model of human coronavirus infection.
天然免疫细胞在机体对多种病原体挑战产生有效的宿主反应中发挥着至关重要的作用。中性粒细胞和单核细胞 - 巨噬细胞等髓系细胞是主要的天然白细胞,它们协调针对病毒性肺部感染的保护性免疫。然而,细胞因子反应失调会促进中性粒细胞和单核细胞 - 巨噬细胞的过度浸润和强烈的促炎活性,导致致命疾病。病毒感染后,浸润的中性粒细胞和单核细胞 - 巨噬细胞的有益或有害作用很大程度上取决于它们分泌炎性细胞因子和趋化因子的能力。大多数研究使用浸润细胞的总数及其激活状态作为衡量指标来证明它们在感染过程中的作用。因此,中性粒细胞和炎性单核巨噬细胞(IMMs)分泌炎性细胞因子和趋化因子的能力及其与疾病严重程度的相关性尚未明确界定。在本章中,我们报告了用于识别肺部浸润的天然免疫细胞并确定其激活状态的有用标志物。我们还描述了一种简单的方法来测量细胞内细胞因子的产生,以评估人冠状病毒感染小鼠模型中中性粒细胞和IMMs的炎性活性。