Transient transfection with WT-CFTR rescues SHS impaired bacterial phagocytosis and limits survival.

Abstract

<p>(A) RAW264.7 cells were seeded on a 24-well plate and transiently transfected (using Lipofectamine) with pcDNA3.1 control vector or pcDNA3.1-WTCFTR for 48 hours. Cells were infected with <i>PA01</i>-GFP (MOI 10) and/or treated with cigarette smoke extract (CSE; 10%; SHS model) for 150 min before microscopy. Representative bright field (top) and fluorescent images (bottom) are shown (magnification 40X, n = 4, white bar = 20μm). (B) Quantification of bacterial phagocytosis (from 4A) shows that transfection with WT-CFTR significantly (*p<0.03) rescues SHS impaired phagocytosis. (C) RAW264.7 cells were treated (as described in 4A) and media (100 μL) was collected and spread on 2% LB agar plates. The plates were incubated overnight at 37°C followed by quantification of bacterial survival by colony forming unit (CFU) counts. Transient transfection with WT-CFTR significantly (*p<0.02) limits bacterial survival in the CSE/SHS treated group, but bacterial survival remains higher than the untreated controls suggesting higher CFTR expression may be needed to limit bacterial burden.</p

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