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Analysis of LC3-associated phagocytosis and antigen presentation


Ligeon, Laure-Anne; Romao, Susana; Münz, Christian (2017). Analysis of LC3-associated phagocytosis and antigen presentation. In: Botelho, Roberto. Phagocytosis and Phagosomes. New York: Springer, 145-168.

Abstract

The noncanonical macroautophagy pathway, LC3-associated phagocytosis (LAP) has recently emerged as an important catabolic process involved during exogenous antigen processing. It has been described that in human macrophages and dendritic cells the direct recruitment of LC3 to the phagosomal membrane is associated with its maturation impairment, allowing the stabilization of the cargo to prolong antigen presentation on major histocompatibility complex (MHC) class II molecules. In this chapter, we describe methods to monitor, manipulate, and understand the role of LAP during MHC class II presentation. We show how to enhance LAP formation resulting in antigen presentation by using zymosan or beads coated with Candida albicans extract. Then, we describe how to determine the localization of Rab7 or Lamp2 on LC3-phagosomes by confocal microscopy, a useful technique to follow phagosome maturation. Finally, we propose an assay to understand how MHC class II antigen presentation can be modulated by the LAP pathway.

Abstract

The noncanonical macroautophagy pathway, LC3-associated phagocytosis (LAP) has recently emerged as an important catabolic process involved during exogenous antigen processing. It has been described that in human macrophages and dendritic cells the direct recruitment of LC3 to the phagosomal membrane is associated with its maturation impairment, allowing the stabilization of the cargo to prolong antigen presentation on major histocompatibility complex (MHC) class II molecules. In this chapter, we describe methods to monitor, manipulate, and understand the role of LAP during MHC class II presentation. We show how to enhance LAP formation resulting in antigen presentation by using zymosan or beads coated with Candida albicans extract. Then, we describe how to determine the localization of Rab7 or Lamp2 on LC3-phagosomes by confocal microscopy, a useful technique to follow phagosome maturation. Finally, we propose an assay to understand how MHC class II antigen presentation can be modulated by the LAP pathway.

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Additional indexing

Item Type:Book Section, refereed, further contribution
Communities & Collections:04 Faculty of Medicine > Institute of Experimental Immunology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2017
Deposited On:16 Jan 2017 09:46
Last Modified:14 Mar 2018 15:34
Publisher:Springer
Series Name:Methods in Molecular Biology
Number:1519
ISSN:1064-3745
ISBN:978-1-4939-6579-3
OA Status:Green
Publisher DOI:https://doi.org/10.1007/978-1-4939-6581-6_10
PubMed ID:27815878

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