研究脓毒症天然免疫反应的方法。

Methods to Study the Innate Immune Response to Sepsis.

作者信息

Walker Wendy E

机构信息

Center of Emphasis in Infectious Diseases, Department of Biomedical Sciences, Texas Tech University Health Sciences Center El Paso, El Paso, TX, USA.

出版信息

Methods Mol Biol. 2018;1717:189-206. doi: 10.1007/978-1-4939-7526-6_15.

DOI:10.1007/978-1-4939-7526-6_15

PMID:29468593

Abstract

This chapter describes techniques to measure the innate immune response in the mouse cecal ligation and puncture model of sepsis. The reader will learn how to perform retro-orbital bleeds to harvest serum from mice and learn how to perform peritoneal lavage to harvest cells and inflammatory mediators from this compartment. The enzyme-linked immunosorbent assay (ELISA) technique is described as a method to measure the levels of cytokines and chemokines in these fluids. Additionally, this chapter describes techniques to stain the cellular fraction of the peritoneal lavage with fluorescently labeled antibodies, and perform fluorescence activated cell sorting (FACS) to quantify macrophages and neutrophils in this compartment.

摘要

本章介绍了在小鼠盲肠结扎和穿刺脓毒症模型中测量先天性免疫反应的技术。读者将学习如何进行眶后采血以从小鼠获取血清，以及如何进行腹腔灌洗以从该腔室获取细胞和炎症介质。酶联免疫吸附测定（ELISA）技术被描述为一种测量这些液体中细胞因子和趋化因子水平的方法。此外，本章还介绍了用荧光标记抗体对腹腔灌洗细胞部分进行染色，以及进行荧光激活细胞分选（FACS）以定量该腔室中巨噬细胞和中性粒细胞的技术。