Terminal sialylation is altered in airway cells with impaired CFTR-mediated chloride transport

Department of Pediatrics, Case Western Reserve University at Rainbow Babies and Children's Hospital, Cleveland, Ohio 44106, USA.
AJP Lung Cellular and Molecular Physiology (Impact Factor: 4.08). 04/2001; 280(3):L482-92.
Source: PubMed


Reduced terminal sialylation at the surface of airway epithelial cells from patients with cystic fibrosis may predispose them to bacterial infection. To determine whether a lack of chloride transport or misprocessing of mutant cystic fibrosis transmembrane conductance regulator (CFTR) is critical for the alterations in glycosylation, we studied a normal human tracheal epithelial cell line (9/HTEo(-)) transfected with the regulatory (R) domain of CFTR, which blocks CFTR-mediated chloride transport; DeltaF508 CFTR, which is misprocessed, wild-type CFTR; or empty vector. Reduced cAMP-stimulated chloride transport is seen in the R domain and DeltaF508 transfectants. These two cell lines had consistent, significantly reduced binding of elderberry bark lectin, which recognizes terminal sialic acid in the alpha-2,6 configuration. Binding of other lectins, including Maakia amurensis lectin, which recognizes sialic acid in the alpha-2,3 configuration, was comparable in all cell lines. Because the cell surface change occurred in R domain-transfected cells, which continue to express wild-type CFTR, it cannot be related entirely to misprocessed or overexpressed CFTR. It is associated most closely with reduced CFTR activity.

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    • "Our prior studies, in two separate cell model systems, have shown that there is increase in available asialoGM1 (aGM1), which binds to P. aeruginosa pilin and flagellin and serves as a major ligand for this organism, on the CF member of the cell pair [17-19]. In these same cell pairs, there is an increased response of IL-8, IL-6, and granulocyte macrophage colony stimulating factor (GM-CSF) to a laboratory strain of P. aeruginosa, PAO1, in the CF member of the pair [6]. "
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    ABSTRACT: In many model systems, cystic fibrosis (CF) phenotype airway epithelial cells in culture respond to P. aeruginosa with greater interleukin (IL)-8 and IL-6 secretion than matched controls. In order to test whether this excess inflammatory response results from the reported increased adherence of P. aeruginosa to the CF cells, we compared the inflammatory response of matched pairs of CF and non CF airway epithelial cell lines to the binding of GFP-PAO1, a strain of pseudomonas labeled with green fluorescent protein. There was no clear relation between GFP-PAO1 binding and cytokine production in response to PAO1. Treatment with exogenous aGM1 resulted in greater GFP-PAO1 binding to the normal phenotype compared to CF phenotype cells, but cytokine production remained greater from the CF cell lines. When cells were treated with neuraminidase, PAO1 adherence was equalized between CF and nonCF phenotype cell lines, but IL-8 production in response to inflammatory stimuli was still greater in CF phenotype cells. The polarized cell lines 16HBEo-Sense (normal phenotype) and Antisense (CF phenotype) cells were used to test the effect of disrupting tight junctions, which allows access of PAO1 to basolateral binding sites in both cell lines. IL-8 production increased from CF, but not normal, cells. These data indicate that increased bacterial binding to CF phenotype cells cannot by itself account for excess cytokine production in CF airway epithelial cells, encourage investigation of alternative hypotheses, and signal caution for therapeutic strategies proposed for CF that include disruption of tight junctions in the face of pseudomonas infection.
    Respiratory research 02/2005; 6(1):69. DOI:10.1186/1465-9921-6-69 · 3.09 Impact Factor
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    • "Eine mögliche Erklärung der veränderten terminalen Glycosylierung bei der Cystischen Fibrose ist eine Veränderung des Golgi pH-Wertes bzw. des Trans-Golgi-Netzwerkes (Kube et al., 2001; Scanlin und Glick, 1999; Tatterson et al., 2001). Der Golgi pH- Wert liegt in der Regel unter dem des Zytoplasmas zwischen pH 6,2 und pH 6,4 (al-Awqati, 1995; Barasch und al-Awqati, 1993). "
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    ABSTRACT: Hamburg, Univ., Diss., 2004. Computerdatei im Fernzugriff.
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    ABSTRACT: Altered terminal glycosylation, with increased fucosylation and decreased sialylation is a hallmark of the cystic fibrosis (CF) glycosylation phenotype. Oligosaccharides purified from the surface membrane glycoconjugates of CF airway epithelial cells have the Lewis x, selectin ligand in terminal positions. This review is focused on the investigations of the glycoconjugates of the CF airway epithelial cell surface. Two of the major bacterial pathogens in CF, Pseudomonas aeruginosa and Haemophilus influenzae, have binding proteins which recognize fucose in alpha-1,3 linkage and asialoglycoconjugates. Therefore, consideration has been given to the possibility that the altered terminal glycosylation of airway epithelial glycoproteins in CF contributes to both the chronic infection and the robust, but ineffective, inflammatory response in the CF lung. Since the glycosylation phenotype of CF airway epithelial cells have been modulated by the expression of wtCFTR, the hypotheses which have been proposed to relate altered function of CFTR to the regulation of the glycosyltransferases are discussed. Understanding the effects of mutant CFTR on glycosylation may provide further insight into the regulation of glycoconjugate processing as well as new approaches to the therapy of CF.
    Glycoconjugate Journal 10/2001; 18(9):649-59. DOI:10.1023/A:1020815205022 · 2.52 Impact Factor
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