Water-pipe smoke condensate increases the internalization of Mycobacterium Bovis of type II alveolar epithelial cells (A549)
Authors: Ian M Adcock
Jahangir Ghorbani
Esmaeil Mortaz
Masoud Movassaghi
Shamila D Alipoor
Johan Garssen
Gert Folkerts
Mohammad Varahram
Journal: BMC Pulmonary Medicine (2017)
Discipline: CLINICAL MEDICINE
Keywords: Endocytosis Activity; Type II Alveolar Epithelial Cells (A549); Water pipe.
Affiliations: Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran., Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Institute of Medical Biotechnology, Molecular Medicine Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran., Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran., Department of Pathology and Laboratory Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA. ., Mycobacteriology Research Center (MRC) National Research Institute of Tuberculosis and lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran., Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Utrecht, The Netherlands., Nutricia Research Centre for Specialized Nutrition, Utrecht, The Netherlands., Cell and Molecular Biology Group, Airways Disease Section, National Heart and Lung Institute, Imperial College London, Dovehouse Street, London, UK., Priority Research Centre for Healthy Lungs, Hunter Medical Research Institute, The University of Newcastle, Newcastle, New South Wales, Australia.

Abstract:  Background: Tuberculosis (TB) is a major global health problem, and there is an association between tobacco smoke and TB. Water pipe smoking has become an increasing problem not only in Middle Eastern countries but also globally because users consider it as safer than cigarettes. The presence of high levels of toxic substances in water-pipe smoke may be a predisposing factor that enhances the incidence of pulmonary disorders. For example, uncontrolled macropinocytosis in alveolar epithelial cells following exposure to water-pipe smoke may predispose subjects to pulmonary infection. Here, we studied the effects of water-pipe condense (WPC) on the internalization of Mycobacterium Bovis BCG by macropinocytosis in the alveolar epithelial cell line A549. Methods: A549 cells were exposed to WPC (4 mg/ml) for 24, 48, 72 and 96 h. Cell viability was studied using the methyl thiazolyldipenyl-tetrazolium bromide (MTT) reduction assay and proliferation by bromodeoxyUridine (BrdU) incorporation. Cells were exposed to FITC-Dextran (1 mg/ml) (as a control) and FITC-BCG (MOI = 10) for 20 min at 37 °C before cells were collected and the uptake of BCG-FITC determined by flow cytometry. Similar experiments were performed at 4 °C as a control. The Rho-associated protein kinase (ROCK) inhibitor Y-27632 (1 μM) was used to assess the mechanism by which WPC enhanced BCG uptake. Results: WPC (4 mg/ml) increased the uptake of BCG-FITC after 72 (1.3 ± 0.1 fold, p &lt; 0.05) and 96 (1.4 ± 0.05 fold, p &lt; 0.05) hours. No effect on BCG-FITC uptake was observed at 24 or 48 h. WPC also significantly increased the uptake of FITC-Dextran (2.9 ± 0.3 fold, p &lt; 0.05) after 24 h. WPC significantly decreased cell viability after 24 (84 ± 2%, p &lt; 0.05), 48 (78±, 3%, p &lt; 0.05), 72 (64 ± 2%, p &lt; 0.05) and 96 h (45 ± 2%, p &lt; 0.05). Y-27632 completely attenuated the increased uptake of BCG by WPC. Cell proliferation showed a decreasing trend in a time-dependent manner with WPC exposure. Conclusion: WPC exposure increased epithelial cell endocytosis activity and death as well as enhancing their capacity for macropinocytosis. Our in vitro data indicates possible harmful effects of WPC on the ability of lung epithelial cells to phagocytose mycobacterium. 

Retracted: No

PubPeer comments:
 The flow cytometry results presented in figures 4a and 4c appear unexpectedly similar. Could the authors please examine the original data and comment? *** ![file] *** Full figure ![file] , 
 Thank you for noticing this. I have contacted the lead author and will get back to you regarding this. BW, Ian , 
 Dr Alipoor has now sent the correct plots for Figure 4. At some point during the many revisions of the paper, one of the plots must have been duplicated. We apologise for this error. The correct plots are below and I have contacted the Journal regarding this. BW, Ian ![file] , 
 2020 correction. https: bmcpulmmed.biomedcentral.com articles 10.1186 s12890-020-01244-1 Correction to: BMC Pulm Med 17, 68 (2017) https: doi.org 10.1186 s12890-017-0413-7 Following publication of the original article [1], the authors that there is an error in the FACs plots detailed in Fig. 4. The error is that the plot of panel u2018Cu2019 has been duplicated as the plot of panel u2018Au2019. Please see the corrected figure in this correction article. The authors apologize for any inconvenience caused. ,