Abstract
Macrophage migration inhibitory factor (MIF) is an upstream regulator of innate immunity and dysregulated MIF is a key mediator of acute and chronic inflammatory processes, autoimmune and cardiovascular diseases, as well as cancer. MIF is a pleiotropic cytokine with chemokine-like functions that has been designated as an atypical chemokine (ACK). It orchestrates leukocyte recruitment and migration into inflamed tissues through non-cognate interactions with the classical chemokine receptors CXCR2 and CXCR4, pathways that are further facilitated by MIF’s cognate receptor CD74. Here, we describe two complementary methods that can be used to characterize immune cell migration and motility responses controlled by MIF and its receptors. These are the Transwell filter migration assay, also known as modified Boyden chamber assay, a two-dimensional (2D) device, and a matrix-based three-dimensional (3D) chemotaxis assay. The Transwell system is primarily suitable to study chemotactic cell transmigration responses toward a chemoattractant such as MIF through a porous filter membrane. The 3D chemotaxis setup enables for the cellular tracking of migration, invasion, and motility of single cells using live cell imaging.
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Acknowledgements
This work was supported by the Metiphys scholarship of the Ludwig-Maximilians-University (LMU) Munich to A.H. and by Deutsche Forschungsgemeinschaft (DFG) grants SFB1123-A03, BE 1977/10-1, and BE 1977/11-1 to J.B. It was co-supported by the Wilhelm-Sander-Stiftung grant 2017.009.1 and the Else-Kröner-Fresenius-Stiftung (EKFS) grant 2014/A216 to J.B. as well as by DFG under Germany’s Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy—ID 390857198) to J.B. We thank ibidi GmbH (Martinsried, Germany) for providing us with the original figures illustrating the 3D μ-Slide chemotaxis assay.
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Hoffmann, A., Zwißler, L.C., El Bounkari, O., Bernhagen, J. (2020). Studying the Pro-Migratory Effects of MIF. In: Harris, J., Morand, E. (eds) Macrophage Migration Inhibitory Factor. Methods in Molecular Biology, vol 2080. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9936-1_1
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DOI: https://doi.org/10.1007/978-1-4939-9936-1_1
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