Methods to Study the Innate Immune Response to Sepsis
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.
Key wordsSepsis CLP Innate immune response Inflammatory response Retro-orbital bleed Peritoneal lavage Enzyme-linked immunosorbent assay (ELISA) Fluorescence-activated cell sorting (FACS) Cytokines Macrophages Neutrophils