Abstract
Numerous immune receptors have the ability to mediate phagocytosis of large particles by triggering dynamic local rearrangement of the cytoskeleton and cell membrane. Different receptors can be differentially recruited to sites of particle binding, which in turn can have important functional consequences with respect to engulfment and downstream signaling. Using Fcγ receptor-mediated phagocytosis of IgG-coated particles as a model, we describe a method for analyzing nascent phagocytic cups and quantifying relative receptor levels at sites of phagocytosis. This technique is based on a ratiometric analysis of subcellular localization of exogenously expressed receptors carrying different fluorescent protein tags. This approach could be applied more generally to the analysis of surface membrane protein localization in the context of any dynamic cellular process.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Greenberg, S. and Grinstein, S. (2002) Phagocytosis and innate immunity. Curr Opin Immunol 14, 136–145.
Swanson, J. A. (2008) Shaping cups into phagosomes and macropinosomes. Nat Rev Mol Cell Biol. 9, 639–649.
Syam, S., Mero, P., Pham, T., McIntosh, C. A., Bruhns, P., and Booth, J. W. (2010) Differential recruitment of activating and inhibitory Fc gamma RII during phagocytosis. J Immunol 184, 2966–2973.
Nimmerjahn, F. and Ravetch, J. V. (2008) Fcgamma receptors as regulators of immune responses. Nat Rev Immunol 8, 34–47.
Henry, R. M., Hoppe, A. D., Joshi, N., and Swanson, J. A. (2004) The uniformity of phagosome maturation in macrophages. J Cell Biol 164, 185–194.
Indik, Z., Kelly, C., Chien, P., Levinson, A. I., and Schreiber, A. D. (1991) Human Fc gamma RII, in the absence of other Fc gamma receptors, mediates a phagocytic signal. J Clin Invest 88, 1766–1771.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Mero, P., Booth, J.W.D. (2011). Ratiometric Analysis of Subcellular Recruitment of Fc Receptors During Phagocytosis. In: Rast, J., Booth, J. (eds) Immune Receptors. Methods in Molecular Biology, vol 748. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-139-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-139-0_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-138-3
Online ISBN: 978-1-61779-139-0
eBook Packages: Springer Protocols