Abstract
Moraxella catarrhalis is a major cause of infectious exacerbations of chronic obstructive lung disease. Adhesion of this pathogen to epithelial cells is critical for its pathogenicity. Although much work has been done on identifying surface molecules of M. catarrhalis as adhesins, several adhesion assays were used in these studies which has never been validated or compared to each other. In the present study, we have examined the capacity of M. catarrhalis to adhere to different human epithelial cells. By using the two most commonly used adhesion assays based on the enumeration of colony-forming units or on the counting of adherent bacteria per epithelial cell by light microscopy, we identified significant limitations of both methods. These arose either from differences in strain-specific adhesion pattern on the epithelial cell surface or the dependence on the state of confluence of the epithelial cell layer. We developed a new fluorescence-based adhesion assay and compared our results to the two conventional methods. We demonstrated that the fluorescence-based adhesion assay offers a reliable and convenient method for the quantification of M. catarrhalis adhesion to confluent epithelial cell monolayers.
Similar content being viewed by others
References
Barnes PJ (2003) Chronic obstructive pulmonary disease: molecular and cellular mechanisms. Eur Respir J 22(4):672–688
Boyle FM, Georghiou PR, Tilse MH, McCormack JG (1991) Branhamella (Moraxella) catarrhalis: pathogenic significance in respiratory infections. Med J Aust 154:592
Catlin BW (1990) Branhamella-Catarrhalis—an organism gaining respect as a pathogen. Clin Microbiol Rev 3:293
Aebi C, Lafontaine ER, Cope LD, Latimer LD, Lumbley SL, McCracken SL Jr, Hansen EJ (1998) Phenotypic effect of isogenic uspa1 and uspa2 mutations on Moraxella catarrhalis 035E. Infect Immun 66:3113
Forsgren A, Brant M, Karamehmedovic M, Riesbeck K (2003) The immunoglobulin D-binding protein MID from Moraxella catarrhalis is also an adhesin. Infect Immun 71:3302
Hill DJ, Virji M (2003) A novel cell-binding mechanism of Moraxella catarrhalis ubiquitous surface protein UspA: specific targeting of the N-domain of carcinoembryonic antigen-related cell adhesion molecules by UspA1. Mol Microbiol 48:117
Holm MM, Vanlerberg SL, Sledjeski DD, Lafontaine ER (2003) The hag protein of Moraxella catarrhalis strain O35E is associated with adherence to human lung and middle ear cells. Infect Immun 71:4977
Holm MM, Vanlerberg SL, Foley IM, Sledjeski DD, Lafontaine ER (2004) The Moraxella catarrhalis porin-like outer membrane protein CD is an adhesin for human lung cells. Infect Immun 72:1906
Hu WG, Chen J, McMichael JC, Gu XX (2001) Functional characteristics of a protective monoclonal antibody against serotype A and C lipooligosaccharides from Moraxella catarrhalis. Infect Immun 69:1358
Lafontaine ER, Cope LD, Aebi C, Latimer JL, McCracken GH, Hansen EJ, (2000) The UspA1 protein and a second type of UspA2 protein mediate adherence of Moraxella catarrhalis to human epithelial cells in vitro. J Bacteriol 182:1364
Pearson MM, Lafontaine ER, Wagner NJ, Geme JW III, Hansen EJ (2002) A hag mutant of Moraxella catarrhalis strain O35E is deficient in hemagglutination, autoagglutination, and immunoglobulin D-binding activities. Infect Immun 70:4523
Timpe JM, Holm MM, Vanlerberg SL, Basrur V, Lafontaine ER (2003) Identification of a Moraxella catarrhalis outer membrane protein exhibiting both adhesin and lipolytic activities. Infect Immun 71:4341
Le Blay G, Fliss I, Lacroix C (2004) Comparative detection of bacterial adhesion to Caco-2 cells with ELISA, radioactivity and plate count methods. J Microbiol Meth 59:211
Lewin A, Freytag B, Meister B, Sharbati-Tehrani S, Schafer H, Appel B (2003) Use of a quantitative TaqMan-PCR for the fast quantification of mycobacteria in broth culture, eukaryotic cell culture and tissue. J Vet Med Ser B Infect Dis Vet Public Health 50:505
Rahman I, Shahamat M, Kirchman PA, Russek-Cohen E, Colwell RR (1994) Methionine uptake and cytopathogenicity of viable but nonculturable Shigella dysenteriae type 1. Appl Environ Microbiol 60:3573
Reddel RR, Ke Y, Gerwin BI, Mcmenamin MG, Lechner JF, Su RT, Brash DE, Park JB, Rhim JS, Harris CC (1988) Transformation of human bronchial epithelial-cells by infection with Sv40 Or adenovirus-12 Sv40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing Sv40 early region genes. Cancer Res 48:1904
Cope LD, Lafontaine ER, Slaughter CA, Hasemann CA, Aebi C, Henderson FW, McCracken GH, Hansen EJ (1999) Characterization of the Moraxella catarrhalis uspA1 uspA2 genes and their encoded products. J Bacteriol 181:4026
Zhang P, Chomel BB, Schau MK, Goo JS, Droz S, Kelminson KL, George SS, Lerche NW, Koehler JE (2004) A family of variably expressed outer-membrane proteins (Vomp) mediates adhesion and autoaggregation in Bartonella quintana. Proc NaH Acad Sci USA 101:13630
Bieber D, Ramer SW, Wu CY, Murray WJ, Tobe T, Fernandez R, Schoolnik GK (1998) Type IV Pili, transient bacterial aggregates, and virulence of enteropathogenic Escherichia coli. Science 280:2114
Lafontaine ER, Wall D, Vanlerberg SL, Donabedian H, Sledjeski DD (1995) Moraxella catarrhalis coaggregates with Streptococcus pyogenes and modulates interactions of S. pyogenes with human epithelial cells. Infect Immun 72(11):6689–6693
Erickson HP (2001) Cytoskeleton: evolution in bacteria. Nature 413:30
Ermert L, Bruckner H, Walmrath D, Grimminger F, Aktories K, Suttorp N, Duncker HR, Seeger W (1995) Role of endothelial cytoskeleton in high-permeability edema due to botulinum C-2 toxin in perfused rabbit lungs. Am J Physiol-Lung Cellular Mol Physiol 12:L753
McMichael JC, Green BA (2003) Vaccines for Moraxella catarrhalis and non-typeable haemophilus influenzae. Curr Opin Investig Drugs 4:953
Murphy TF, Sethi S, Niederman MS (2000) The role of bacteria in exacerbations of COPD—a constructive view. Chest 118:204
Sethi S (2000) Infectious etiology of acute exacerbations of chronic bronchitis. Chest 117:380S
Sethi S, Murphy TF (2001) Bacterial infection in chronic obstructive pulmonary disease in 2000: a state-of-the-art review. Clin Microbiol Rev 14:336
Sethi S, Evans N, Grant BJB, Murphy TF (2002) New strains of bacteria and exacerbations of chronic obstructive pulmonary disease. N Engl J Med 347:465
Diebold Y, Calonge M, de Salamanca AE, Callejo S, Corrales RM, Saez V, Siemasko KF, Stern ME (2003) Characterization of a spontaneously immortalized cell line (IOBA-NHC) from normal human conjunctiva. Invest Ophthalmol Vis Sci 44:4263
Masters JR (2002) HeLa cells 50 years on: the good, the bad and the ugly. Nat Rev Cancer 2:315
Saint Jean M, Baudouin C, Di Nolfo M, Roman S, Lozato P, Warnet JM, Brignole F (2004) Comparison of morphological and functional characteristics of primary-cultured human conjunctival epithelium and of Wong-Kilbourne derivative of Chang conjunctival cell line. Exp Eye Res 78:257
Rikitomi N, Ahmed K, Nagatake T (1997) Moraxella (Branhamella) catarrhalis adherence to human bronchial and oropharyngeal cells: the role of adherence in lower respiratory tract infections. Microbiol Immunol 41:487
Ben Ze’ev A (1997) Cytoskeletal and adhesion proteins as tumor suppressors. Curr Opin Cell Biol 9:99
Singer BB, Scheffrahn I, Obrink B (2000) The tumor growth-inhibiting cell adhesion molecule CEACAM1 (C-CAM) is differently expressed in proliferating and quiescent epithelial cells and regulates cell proliferation. Cancer Res 60:1236
Qualtrough D, Hinoi T, Fearon E, Paraskeva C (2002) Expression of CDX2 in normal and neoplastic human colon tissue and during differentiation of an in vitro model system. Gut 51:184
Hidaka, Nagao T, Asoh A, Kondo Y, Nagao K (1998) Expression of E-cadherin, alpha-catenin, beta-catenin, and gamma-catenin in bronchioloalveolar carcinoma and conventional pulmonary adenocarcinoma: an immunohistochemical study. Mod Pathol 11:1039
Acknowledgements
The technical assistance of Kerstin Möhr and Sylwia Schapke is greatly appreciated. Part of this work will be included in the doctoral thesis of Krishna N. Tiwari. This work was supported in part by grant from the Rahel-Hirsch Stipendium of the Charité University Hospital, Humboldt University, Berlin, Germany.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Slevogt, H., Tiwari, K.N., Schmeck, B. et al. Adhesion of Moraxella catarrhalis to human bronchial epithelium characterized by a novel fluorescence-based assay. Med Microbiol Immunol 195, 73–83 (2006). https://doi.org/10.1007/s00430-005-0003-9
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00430-005-0003-9