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Analysis of LC3-Associated Phagocytosis and Antigen Presentation

Laure-Anne Ligeon1, Susana Romao1, Christian Münz2

  • 1Viral Immunobiology, Institute of Experimental Immunology, University of Zürich, Winterthurerstrasse 190, Zürich, 8057, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)
|November 6, 2016
PubMed

Insights

LC3-associated phagocytosis (LAP) enhances antigen presentation by stabilizing cargo in phagosomes, crucial for MHC class II presentation. This study details methods to monitor and manipulate LAP for improved antigen processing insights.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • LC3-associated phagocytosis (LAP) is a noncanonical macroautophagy pathway critical for exogenous antigen processing.
  • Direct LC3 recruitment to phagosomes impairs maturation, prolonging antigen presentation on MHC class II molecules in human cells.

Purpose of the Study:

  • To describe methods for monitoring, manipulating, and understanding LAP's role in MHC class II antigen presentation.
  • To provide techniques for enhancing LAP formation and assessing its impact on antigen presentation.

Main Methods:

  • Enhancement of LAP formation using zymosan or Candida albicans extract-coated beads.
  • Confocal microscopy to determine Rab7 or Lamp2 localization on LC3-phagosomes for phagosome maturation tracking.
  • Development of an assay to investigate LAP pathway modulation of MHC class II antigen presentation.

Main Results:

  • Demonstrated methods to enhance LAP formation, leading to increased antigen presentation.
  • Established techniques to visualize phagosome maturation markers (Rab7, Lamp2) in relation to LC3-phagosomes.
  • Proposed a novel assay to study the modulation of MHC class II antigen presentation by LAP.

Conclusions:

  • LAP plays a significant role in modulating MHC class II antigen presentation.
  • The described methods provide valuable tools for studying LAP function in antigen processing and presentation.
  • Understanding LAP is key to developing strategies for enhancing immune responses via antigen presentation.

Keywords:
AutophagyConfocal microscopyLC3-associated phagosomeMHC class IIMHC class II antigen presentationPhagosome assayPhagosome maturation

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