Pseudomonas aeruginosa internalization by human epithelial respiratory cells depends on cell differentiation, polarity, and junctional complex integrity

被引:76
作者
Plotkowski, MC
de Bentzmann, S
Pereira, SHM
Zahm, JM
Bajolet-Laudinat, O
Roger, P
Puchelle, E
机构
[1] State Univ Rio de Janeiro, Dept Microbiol & Immunol, Rio De Janeiro, Brazil
[2] INSERM, U314, F-75654 Paris 13, France
[3] INSERM, IFR 53, F-75654 Paris, France
[4] Hop Robert Debre, Lab Bacteriol Virol Hyg, Reims, France
关键词
D O I
10.1165/ajrcmb.20.5.3408
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Internalization of Pseudomonas aeruginosa by epithelial respiratory cell lines has been suggested to be dependent on the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Because we have observed intracellular (IC) P. aeruginosa only in cells that do not express apical CFTR, we addressed the question of whether bacterial internalization by epithelial cells depends on the degree of cell differentiation and polarity. Internalization of piliated P, aeruginosa PAO-1 and PAK by human epithelial respiratory cells in primary culture and by the 16 human bronchial epithelial 140- cell line cultured either on thick collagen gels or on thin collagen films was evaluated by the gentamicin exclusion assay. Cells cultured on thick gels were differentiated, polarized, and tight. They exhibited CFTR at their apical membranes, expressed beta 1 integrins at their basal membranes, excluded lanthanum nitrate, and uniformly expressed ZO-1 protein. In contrast, in cells cultured on thin films, CFTR was present mainly in the cytoplasm, whereas beta 1 integrins were detected at epical membranes. Most cells cultured on thin films did not exclude lanthanum nitrate and rarely expressed ZO-1 protein. Cells grown on thick and thin collagen substrates differed markedly in bacterial internalization: no IC bacteria could be detected in cells cultured on gels, whereas high IC bacterial concentrations were isolated from cells cultured on thin films. Treatment of cells cultured on thin films with ethylenediaminetetraacetic acid, to disrupt intercellular junctions further, significantly enhanced P. aeruginosa internalization. Our results suggest that P. aeruginosa internalization by epithelial respiratory cells does not depend on CFTR protein expression at the epithelial cell surface but rather on cell polarity and junctional complex integrity.
引用
收藏
页码:880 / 890
页数:11
相关论文
共 38 条
[1]   INFLUENCE OF COLLAGEN GEL ON THE ORIENTATION OF EPITHELIAL-CELL POLARITY - FOLLICLE FORMATION FROM ISOLATED THYROID-CELLS AND FROM PREFORMED MONOLAYERS [J].
CHAMBARD, M ;
GABRION, J ;
MAUCHAMP, J .
JOURNAL OF CELL BIOLOGY, 1981, 91 (01) :157-166
[2]   DEFECTIVE INTRACELLULAR-TRANSPORT AND PROCESSING OF CFTR IS THE MOLECULAR-BASIS OF MOST CYSTIC-FIBROSIS [J].
CHENG, SH ;
GREGORY, RJ ;
MARSHALL, J ;
PAUL, S ;
SOUZA, DW ;
WHITE, GA ;
ORIORDAN, CR ;
SMITH, AE .
CELL, 1990, 63 (04) :827-834
[3]  
CHEVILLARD M, 1993, EPITHELIAL CELL BIOL, V2, P17
[4]  
COZZENS AL, 1992, IN VITRO CELL DEV, V28, P735
[5]   Receptors in the Pseudomonas aeruginosa adherence to injured and repairing airway epithelium [J].
deBentzmann, S ;
Plotkowski, C ;
Puchelle, E .
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 1996, 154 (04) :S155-S162
[6]   PROTECTION OF HUMAN RESPIRATORY EPITHELIUM FROM PSEUDOMONAS-AERUGINOSA ADHERENCE BY PHOSPHATIDYLGLYCEROL LIPOSOMES [J].
DEBENTZMANN, SG ;
BAJOLETLAUDINAT, O ;
DUPUIT, F ;
PIERROT, D ;
FUCHEY, C ;
PLOTKOWSKI, MC ;
PUCHELLE, E .
INFECTION AND IMMUNITY, 1994, 62 (02) :704-708
[7]   RAPID COLORIMETRIC ASSAY FOR CELL-GROWTH AND SURVIVAL - MODIFICATIONS TO THE TETRAZOLIUM DYE PROCEDURE GIVING IMPROVED SENSITIVITY AND RELIABILITY [J].
DENIZOT, F ;
LANG, R .
JOURNAL OF IMMUNOLOGICAL METHODS, 1986, 89 (02) :271-277
[8]  
DIMANGO E, 1996, PED PULMON S, V13, P287
[9]   CFTR AND DIFFERENTIATION MARKERS EXPRESSION IN NON-CF AND DELTA-F-508 HOMOZYGOUS CF NASAL EPITHELIUM [J].
DUPUIT, F ;
KALIN, N ;
BREZILLON, S ;
HINNRASKY, J ;
TUMMLER, B ;
PUCHELLE, E .
JOURNAL OF CLINICAL INVESTIGATION, 1995, 96 (03) :1601-1611
[10]  
FALK P, 1994, MICROBES TOOLS CELL, P165