Summary
Macrophage activation can be divided into a classical and an alternative pathway. Interferon-gamma-induced, classically activated macrophages are indispensable for protective effector responses against intracellular pathogens. However, excessive inflammatory immune responses mediated by classical macrophage activation can also be detrimental to the host. In contrast, the IL-4 receptor-alpha-mediated alternative pathway of macrophage activation has been proposed as a mechanism to attenuate excessive inflammation. Indeed, the generation of macrophage/neutrophil-specific IL-4 receptor-alpha-deficient mice (LysMcreIL-4Rαα-/lox) enables us now to evaluate the importance of this type of macrophage activation in vivo. Thus, the analysis of LysMcreIL-4Rα−/lox mice and the phenotypic characterization of macrophage activation during inflammatory immune responses become of major importance for inflammation research, and useful markers have been identified that allow classically and alternatively activated macrophages to be distinguished. Inducible nitric oxide synthase and arginase-1 are not only prototypical markers of classical and alternative macrophage activation, but both enzymes are also strongly involved in regulating macrophage effector mechanisms and inflammatory immune responses. In this chapter, we describe the use of LysMcreIL-4Rα−/lox mice and present experimental procedures to determine classical versus alternative macrophage activation by analyzing nitric oxide synthase and arginase-1 in vitro and in vivo in this murine model.
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Acknowledgments
Related projects are supported by grants to FB (The Wellcome Trust and Royal Society, UK; German Research Foundation, Germany; National Research Foundation and Medical Research Council, ZA) and CH (University of Lübeck, Germany, Research Focus “Host Defense against Infectious Diseases”, Project B2 “Arginase-1 mediated modulation of granuloma necrosis and immune defense against mycobacterial infections”; Nation-wide Collaborative Grant: “Pulmonary Tuberculosis – host and pathogen determinants of resistance and disease progression.” Workpackages E (Animal models).
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Brombacher, F., Arendse, B., Peterson, R., Hölscher, A., Hölscher, C. (2009). Analyzing Classical and Alternative Macrophage Activation in Macrophage/Neutrophil-Specific IL-4 Receptor-Alpha-Deficient Mice. In: Reiner, N. (eds) Macrophages and Dendritic Cells. Methods in Molecular Biology™, vol 531. Humana Press. https://doi.org/10.1007/978-1-59745-396-7_15
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DOI: https://doi.org/10.1007/978-1-59745-396-7_15
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