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Cholesterol is obligatory for polarization and chemotaxis but not for endocytosis and associated signaling from chemoattractant receptors in human neutrophils

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

Plasma membrane cholesterol is critical for neutrophil chemotaxis, although how cholesterol affects chemotactic signaling pathway has not been clearly delineated. Here we demonstrate that cholesterol was absolutely required for polarized redistribution of key chemotactic mediators in human neutrophils in response to all chemoattractants tested (fMet-Leu-Phe, and the chemokines CXCL1, CXCL8 and CXCL12). In particular, PI3K and phosphatidylinositol-3,4,5 triphosphate (PIP3) failed to accumulate at the front and phosphatase and tensin homolog (PTEN) at the back of chemoattractant-stimulated neutrophils after cholesterol depletion. Cholesterol depletion did not affect early chemoattractant signaling events such as G-protein activation, intracellular calcium flux or G-protein-independent endocytosis-linked signaling, including the activation of mitogen-activated protein kinase (MAPK), Hck and Fgr transduced by β-arrestin. During cell polarization, F-actin assemblies redistributed the cholesterol-rich microdomains and cytoskeleton-anchored proteins, including CD16 and CD44 from the leading edge. These data suggest that spatial polarization of chemotactic mediators is orchestrated by protein:protein interactions that organize cholesterol-rich domains of the plasma membrane.

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Abbreviations

APC:

Allophycocyanin

DRMs:

Detergent-resistant membranes

EGFR:

Epidermal growth factor receptor

fMLF:

N-Formyl-methionine-leucine-phenylalanine

FPR:

N-Formyl peptide receptor

GPCR:

G-protein-coupled receptor

GPI:

Glycosyl phosphatidylinositol

GRK:

G-protein-coupled receptor kinase

GROα:

Growth related oncogene α

HMG-CoA:

Hydroxy-methyl glutaryl CoA

IL-8:

Interleukin-8

IP3 :

Inositol-1,4,5 trisphosphate

Lat-B:

Latrunculin B

LTB4 :

Leukotriene B4

MAPK:

Mitogen-activated protein kinase

MβCD:

Methyl-β-cyclodextrin

PIP2 :

Phosphatidyl inositol 4,5 bisphosphate

PIP3 :

Phosphatidylinositol-3,4,5 triphosphate

PTEN:

Phosphatase and tensin homolog

PTX:

Pertussis toxin

SDF:

Stromal cell-derived factor

TM:

Transmembrane

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Acknowledgements

We thank Philip Murphy and Joshua Farber of LMI, NIAID for advice and comments and Owen Schwartz of the Biological Imaging Section, RTB, NIAID for technical advice. The Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health supported this research.

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  1. Laboratory of Molecular Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 10, Room 6A05, Bethesda, MD, 20892-1576, USA

    Jeremy J. Rose, John F. Foley, Ling Yi, Gina Herren & Sundararajan Venkatesan

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  1. Jeremy J. Rose
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  2. John F. Foley
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  3. Ling Yi
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Correspondence to Sundararajan Venkatesan.

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Rose, J.J., Foley, J.F., Yi, L. et al. Cholesterol is obligatory for polarization and chemotaxis but not for endocytosis and associated signaling from chemoattractant receptors in human neutrophils. J Biomed Sci 15, 441–461 (2008). https://doi.org/10.1007/s11373-008-9239-x

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  • DOI: https://doi.org/10.1007/s11373-008-9239-x

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