Second-hand cigarette smoke impairs bacterial phagocytosis in macrophages by modulating CFTR dependent lipid-rafts

PloS One
Inzer NiNeeraj Vij

Abstract

First/Second-hand cigarette-smoke (FHS/SHS) exposure weakens immune defenses inducing chronic obstructive pulmonary disease (COPD) but the underlying mechanisms are not fully understood. Hence, we evaluated if SHS induced changes in membrane/lipid-raft (m-/r)-CFTR (cystic fibrosis transmembrane conductance regulator) expression/activity is a potential mechanism for impaired bacterial phagocytosis in COPD. RAW264.7 murine macrophages were exposed to freshly prepared CS-extract (CSE) containing culture media and/or Pseudomonas-aeruginosa-PA01-GFP for phagocytosis (fluorescence-microscopy), bacterial survival (colony-forming-units-CFU), and immunoblotting assays. The CFTR-expression/activity and lipid-rafts were modulated by transient-transfection or inhibitors/inducers. Next, mice were exposed to acute/sub-chronic-SHS or room-air (5-days/3-weeks) and infected with PA01-GFP, followed by quantification of bacterial survival by CFU-assay. We investigated the effect of CSE treatment on RAW264.7 cells infected by PA01-GFP and observed that CSE treatment significantly (p<0.01) inhibits PA01-GFP phagocytosis as compared to the controls. We also verified this in murine model, exposed to acute/sub-chronic-SHS and found significant (p<0.05...Continue Reading

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Citations

Dec 9, 2015·BMC Pulmonary Medicine·Borja G CosíoAlvar Agustí
Jul 13, 2016·Journal of Immunology Research·William G BainNeil R Aggarwal
Aug 27, 2016·Expert Opinion on Drug Delivery·Neeraj Vij
Feb 6, 2017·Molecular and Cellular Pediatrics·Neel PatelNeeraj Vij
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May 6, 2017·Respiratory Research·Manish Bodas, Neeraj Vij
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Apr 9, 2020·Expert Review of Respiratory Medicine·Rhonda SzczesniakJoseph M Collaco
Oct 30, 2020·International Journal of Chronic Obstructive Pulmonary Disease·Steven M RoweSarah S Grant
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May 18, 2021·Mechanisms of Ageing and Development·Juliana Ruiz FernandesGil Benard
Jul 22, 2021·American Journal of Physiology. Lung Cellular and Molecular Physiology·Brandon A PaarwaterNelita du Plessis

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Methods Mentioned

BETA
Assay
Protein Assay
fluorescence microscopy
transfection

Software Mentioned

Image
Image J Photoshop
CellSens

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