Frustrated phagocytosis on micro-patterned immune complexes to characterize lysosome movements in live macrophages.

Frontiers in Immunology
Arnaud LabrousseI Maridonneau-Parini

Abstract

Lysosome mobilization is a key cellular process in phagocytes for bactericidal activities and trans-matrix migration. The molecular mechanisms that regulate lysosome mobilization are still poorly known. Lysosomes are hard to track as they move toward phagosomes throughout the cell volume. In order to anticipate cell regions where lysosomes are recruited to, human and RAW264.7 macrophages were seeded on surfaces that were micro-patterned with immune complexes (ICs) as 4 μm-side squares. Distances between IC patterns were adapted to optimize cell spreading in order to constrain lysosome movements mostly in two dimensions. FcΓ receptors triggered local frustrated phagocytosis, frustrated phagosomes appeared as rings of F-actin dots around the IC patterns as early as 5 min after cells made contact with the substratum. Frustrated phagosomes recruited actin-associated proteins (vinculin, paxillin, and gelsolin). The fusion of lysosomes with frustrated phagosomes was shown by the release of beta-hexosaminidase and the recruitment of Lamp1 to frustrated phagosomes. Lysosomes of RAW264.7 macrophages were labeled with cathepsin-D-mCherry to visualize their movements toward frustrated phagosomes. Lysosomes saltatory movements were markedl...Continue Reading

Citations

Dec 30, 2015·Cellular Immunology·Yianzhu LiuMalgorzata Kloc
Apr 6, 2016·Journal of Lipid Research·Abigail S HakaFrederick R Maxfield
Aug 26, 2014·Inflammatory Bowel Diseases·Beckley K DavisIrving C Allen
Jun 20, 2018·Macromolecular Bioscience·Guoying Zhou, Thomas Groth
Feb 6, 2019·International Journal of Molecular Sciences·Erminia MarianiLia Pulsatelli
Oct 31, 2019·Traffic·Frederick R MaxfieldRajesh K Singh
Mar 7, 2020·International Reviews of Immunology·Vijay Kumar
Jun 5, 2020·Science Advances·C Wyatt ShieldsSamir Mitragotri
Apr 20, 2021·Biochimica Et Biophysica Acta. Molecular Cell Research·Anthony J MorganAntony Galione

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Methods Mentioned

BETA
fluorescence microscopy
atomic force microscopy
AFM

Software Mentioned

ImageJ
Image J
GraphPad Prism

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