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Analysis of LC3-Associated Phagocytosis and Antigen Presentation

  • Laure-Anne Ligeon
  • Susana Romao
  • Christian Münz
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1519)

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.

Key words

Autophagy Confocal microscopy LC3-associated phagosome MHC class II MHC class II antigen presentation Phagosome maturation Phagosome assay 

Notes

Acknowledgments

Research in our laboratory is supported by grants from Cancer Research Switzerland (KFS-3234-08-2013), Worldwide Cancer Research (14-1033), KFSPMS and KFSPHHLD of the University of Zurich, the Sobek Foundation, the Swiss Vaccine Research Institute and the Swiss National Science Foundation (310030_162560 and CRSII3_160708).

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Laure-Anne Ligeon
    • 1
  • Susana Romao
    • 1
  • Christian Münz
    • 1
  1. 1.Viral Immunobiology, Institute of Experimental ImmunologyUniversity of ZürichZürichSwitzerland

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