Second-hand Cigarette Smoke Impairs Bacterial Phagocytosis in Macrophages by Modulating CFTR Dependent Lipid-rafts

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2015 Mar 21

PMID 25794013

Citations 33

Authors

Inzer Ni

Changhoon Ji

Neeraj Vij

Affiliations

Soon will be listed here.

Abstract

Introduction: 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.

Methods: 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.

Results: 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, p<0.02) increase in bacterial survival in the SHS-exposed lungs as compared to the room-air controls. Next, we examined the effect of impaired CFTR ion-channel-activity on PA01-GFP infection of RAW264.7 cells using CFTR172-inhibitor and found no significant change in phagocytosis. We also similarly evaluated the effect of a CFTR corrector-potentiator compound, VRT-532, and observed no significant rescue of CSE impaired PA01-GFP phagocytosis although it significantly (p<0.05) decreases CSE induced bacterial survival. Moreover, induction of CFTR expression in macrophages significantly (p<0.03) improves CSE impaired PA01-GFP phagocytosis as compared to the control. Next, we verified the link between m-/r-CFTR expression and phagocytosis using methyl-β-cyclodextran (CD), as it is known to deplete CFTR from membrane lipid-rafts. We observed that CD treatment significantly (p<0.01) inhibits bacterial phagocytosis in RAW264.7 cells and adding CSE further impairs phagocytosis suggesting synergistic effect on CFTR dependent lipid-rafts.

Conclusion: Our data suggest that SHS impairs bacterial phagocytosis by modulating CFTR dependent lipid-rafts.

Citing Articles

Role of mitochondria in inflammatory lung diseases.

Narala V, Narala S, Aiya Subramani P, Panati K, Kolliputi N Front Pharmacol. 2024; 15:1433961.

PMID: 39228517 PMC: 11368744. DOI: 10.3389/fphar.2024.1433961.

Cigarette smoke sustains immunosuppressive microenvironment inducing M2 macrophage polarization and viability in lung cancer settings.

Bianchi F, Le Noci V, Bernardo G, Gagliano N, Colombo G, Sommariva M PLoS One. 2024; 19(5):e0303875.

PMID: 38776331 PMC: 11111031. DOI: 10.1371/journal.pone.0303875.

Rafting on the Evidence for Lipid Raft-like Domains as Hubs Triggering Environmental Toxicants' Cellular Effects.

Marques-da-Silva D, Lagoa R Molecules. 2023; 28(18).

PMID: 37764374 PMC: 10536579. DOI: 10.3390/molecules28186598.

Aiming to Improve Equity in Pulmonary Health: Cystic Fibrosis.

Oates G, Schechter M Clin Chest Med. 2023; 44(3):555-573.

PMID: 37517835 PMC: 10458995. DOI: 10.1016/j.ccm.2023.03.011.

CAGE sequencing reveals CFTR-dependent dysregulation of type I IFN signaling in activated cystic fibrosis macrophages.

Gillan J, Chokshi M, Hardisty G, Hendry S, Prasca-Chamorro D, Robinson N Sci Adv. 2023; 9(21):eadg5128.

PMID: 37235648 PMC: 10219589. DOI: 10.1126/sciadv.adg5128.

References

Li C, Schuetz J, Naren A . Tobacco carcinogen NNK transporter MRP2 regulates CFTR function in lung epithelia: implications for lung cancer. Cancer Lett. 2010; 292(2):246-53. PMC: 2868381. DOI: 10.1016/j.canlet.2009.12.009. View

LAURENZI G, GUARNERI J, ENDRIGA R, Carey J . CLEARANCE OF BACTERIA BY THE LOWER RESPIRATORY TRACT. Science. 1963; 142(3599):1572-3. DOI: 10.1126/science.142.3599.1572. View

Deziel E, Comeau Y, Villemur R . Initiation of biofilm formation by Pseudomonas aeruginosa 57RP correlates with emergence of hyperpiliated and highly adherent phenotypic variants deficient in swimming, swarming, and twitching motilities. J Bacteriol. 2001; 183(4):1195-204. PMC: 94992. DOI: 10.1128/JB.183.4.1195-1204.2001. View

Thomas W, Holt P, Keast D . Cigarette smoke and phagocyte function: effect of chronic exposure in vivo and acute exposure in vitro. Infect Immun. 1978; 20(2):468-75. PMC: 421879. DOI: 10.1128/iai.20.2.468-475.1978. View

Bodas M, Min T, Vij N . Critical role of CFTR-dependent lipid rafts in cigarette smoke-induced lung epithelial injury. Am J Physiol Lung Cell Mol Physiol. 2011; 300(6):L811-20. PMC: 3119127. DOI: 10.1152/ajplung.00408.2010. View

Bomberger J, Ye S, MacEachran D, Koeppen K, Barnaby R, OToole G . A Pseudomonas aeruginosa toxin that hijacks the host ubiquitin proteolytic system. PLoS Pathog. 2011; 7(3):e1001325. PMC: 3063759. DOI: 10.1371/journal.ppat.1001325. View

Vij N, Downey G . The yin and yang of cystic fibrosis transmembrane conductance regulator function: implications for chronic lung disease. Am J Respir Crit Care Med. 2013; 187(2):120-2. PMC: 3570650. DOI: 10.1164/rccm.201211-2011ED. View

Hodge S, Hodge G, Ahern J, Jersmann H, Holmes M, Reynolds P . Smoking alters alveolar macrophage recognition and phagocytic ability: implications in chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol. 2007; 37(6):748-55. DOI: 10.1165/rcmb.2007-0025OC. View

Murphy T, Brauer A, Eschberger K, Lobbins P, Grove L, Cai X . Pseudomonas aeruginosa in chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2008; 177(8):853-60. DOI: 10.1164/rccm.200709-1413OC. View

10.

Bajmoczi M, Gadjeva M, Alper S, Pier G, Golan D . Cystic fibrosis transmembrane conductance regulator and caveolin-1 regulate epithelial cell internalization of Pseudomonas aeruginosa. Am J Physiol Cell Physiol. 2009; 297(2):C263-77. PMC: 2724095. DOI: 10.1152/ajpcell.00527.2008. View

11.

Wellhauser L, Kim Chiaw P, Pasyk S, Li C, Ramjeesingh M, Bear C . A small-molecule modulator interacts directly with deltaPhe508-CFTR to modify its ATPase activity and conformational stability. Mol Pharmacol. 2009; 75(6):1430-8. DOI: 10.1124/mol.109.055608. View

12.

Wang Y, Loo T, Bartlett M, Clarke D . Correctors promote maturation of cystic fibrosis transmembrane conductance regulator (CFTR)-processing mutants by binding to the protein. J Biol Chem. 2007; 282(46):33247-33251. DOI: 10.1074/jbc.C700175200. View

13.

Donnelly L, Barnes P . Defective phagocytosis in airways disease. Chest. 2012; 141(4):1055-1062. DOI: 10.1378/chest.11-2348. View

14.

Minematsu N, Blumental-Perry A, Shapiro S . Cigarette smoke inhibits engulfment of apoptotic cells by macrophages through inhibition of actin rearrangement. Am J Respir Cell Mol Biol. 2010; 44(4):474-82. PMC: 3095920. DOI: 10.1165/rcmb.2009-0463OC. View

15.

Moreau-Marquis S, Stanton B, OToole G . Pseudomonas aeruginosa biofilm formation in the cystic fibrosis airway. Pulm Pharmacol Ther. 2008; 21(4):595-9. PMC: 2542406. DOI: 10.1016/j.pupt.2007.12.001. View

16.

Pier G . Role of the cystic fibrosis transmembrane conductance regulator in innate immunity to Pseudomonas aeruginosa infections. Proc Natl Acad Sci U S A. 2000; 97(16):8822-8. PMC: 34018. DOI: 10.1073/pnas.97.16.8822. View

17.

Sethi S, Mallia P, Johnston S . New paradigms in the pathogenesis of chronic obstructive pulmonary disease II. Proc Am Thorac Soc. 2009; 6(6):532-4. DOI: 10.1513/pats.200905-025DS. View

18.

Hodge S, Hodge G, Scicchitano R, Reynolds P, Holmes M . Alveolar macrophages from subjects with chronic obstructive pulmonary disease are deficient in their ability to phagocytose apoptotic airway epithelial cells. Immunol Cell Biol. 2003; 81(4):289-96. DOI: 10.1046/j.1440-1711.2003.t01-1-01170.x. View

19.

Barnes P . Alveolar macrophages as orchestrators of COPD. COPD. 2006; 1(1):59-70. DOI: 10.1081/COPD-120028701. View

20.

Goldberg J, Pier G . The role of the CFTR in susceptibility to Pseudomonas aeruginosa infections in cystic fibrosis. Trends Microbiol. 2000; 8(11):514-20. DOI: 10.1016/s0966-842x(00)01872-2. View