1887

Abstract

The aim of this study was to investigate the interaction of with three different human cell lines: TCC-SUP (epithelial cells from urinary bladder), HeLa (epithelial cells from cervical carcinoma) and Caco-2 (epithelial cells from colorectal adenocarcinoma). In particular we sought to assess the degree of cell damage and activity reduction induced by adhesion and the role of secreted aspartyl proteinase () gene expression in this process. Two strains were used: the reference strain ATCC 750 and a clinical isolate from urine (U69). The ability of to adhere to a confluent layer of human cells was determined using an adaptation of the crystal violet staining method; cell damage and cell activity inhibition induced by the adhesion of were assessed by measuring lactate dehydrogenase and tetrazolium salt (MTS) reduction, respectively. gene expression was determined by real-time PCR. Both strains were able to adhere to the different human cells, although in a strain- and cell-line-dependent manner. Concerning the cellular response to , the highest inhibition of cell activity was obtained for Caco-2, followed by TCC-SUP and HeLa cells. The highest percentage of cell damage (around 14 %) was observed for TCC-SUP cells in contact with the U69 isolate and for Caco-2 in contact with the reference strain. Real-time PCR analysis revealed a wide range of expression profiles of genes for both strains in contact with the different types of epithelial cells. was the gene expressed at the highest level for both strains in contact with the three human epithelial cell lines. The results highlight that the response of human cells to adhesion, as well as production of SAPs, is dependent on both the strain and the epithelial cell line.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.031195-0
2011-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jmm/60/9/1270.html?itemId=/content/journal/jmm/10.1099/jmm.0.031195-0&mimeType=html&fmt=ahah

References

  1. Bendel C. M. 2003; Colonization and epithelial adhesion in the pathogenesis of neonatal candidiasis. Semin Perinatol 27:357–364 [View Article][PubMed]
    [Google Scholar]
  2. Bendel C. M., Hostetter M. K. 1993; Distinct mechanisms of epithelial adhesion for Candida albicans and Candida tropicalis. Identification of the participating ligands and development of inhibitory peptides. J Clin Invest 92:1840–1849 [View Article][PubMed]
    [Google Scholar]
  3. Biasoli M. S., Tosello M. E., Magaró H. M. 2002; Adherence of Candida strains isolated from the human gastrointestinal tract. Mycoses 45:465–469[PubMed]
    [Google Scholar]
  4. Chakrabarti A., Chatterjee S. S., Rao K. L. N., Zameer M. M., Shivaprakash M. R., Singhi S., Singh R., Varma S. C. 2009; Recent experience with fungaemia: change in species distribution and azole resistance. Scand J Infect Dis 41:275–284 [View Article][PubMed]
    [Google Scholar]
  5. Filler S. G., Sheppard D. C. 2006; Fungal invasion of normally non-phagocytic host cells. PLoS Pathog 2:e129 [View Article][PubMed]
    [Google Scholar]
  6. Galán-Ladero M. A., Blanco M. T., Sacristán B., Fernández-Calderón M. C., Pérez-Giraldo C., Gómez-García A. C. 2010; Enzymatic activities of Candida tropicalis isolated from hospitalized patients. Med Mycol 48:207–210 [View Article][PubMed]
    [Google Scholar]
  7. Jayatilake J. A., Samaranayake Y. H., Cheung L. K., Samaranayake L. P. 2006; Quantitative evaluation of tissue invasion by wild type, hyphal and SAP mutants of Candida albicans, and non-albicans Candida species in reconstituted human oral epithelium. J Oral Pathol Med 35:484–491 [View Article][PubMed]
    [Google Scholar]
  8. Kothavade R. J., Kura M. M., Valand A. G., Panthaki M. H. 2010; Candida tropicalis: its prevalence, pathogenicity and increasing resistance to fluconazole. J Med Microbiol 59:873–880 [View Article][PubMed]
    [Google Scholar]
  9. Krcmery V., Barnes A. J. 2002; Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance. J Hosp Infect 50:243–260 [View Article][PubMed]
    [Google Scholar]
  10. Monod M., Capoccia S., Léchenne B., Zaugg C., Holdom M., Jousson O. 2002; Secreted proteases from pathogenic fungi. Int J Med Microbiol 292:405–419 [View Article][PubMed]
    [Google Scholar]
  11. Naglik J. R., Challacombe S. J., Hube B. 2003; Candida albicans secreted aspartyl proteinases in virulence and pathogenesis. Microbiol Mol Biol Rev 67:400–428 [View Article][PubMed]
    [Google Scholar]
  12. Negri M., Gonçalves V., Silva S., Henriques M., Azeredo J., Oliveira R. 2010a; Crystal violet staining to quantify Candida adhesion to epithelial cells. Br J Microbiol 67:120–125
    [Google Scholar]
  13. Negri M., Martins M., Henriques M., Svidzinski T. I., Azeredo J., Oliveira R. 2010b; Examination of potential virulence factors of Candida tropicalis clinical isolates from hospitalized patients. Mycopathologia 169:175–182 [View Article][PubMed]
    [Google Scholar]
  14. Nucci M., Colombo A. L. 2007; Candidemia due to Candida tropicalis: clinical, epidemiologic, and microbiologic characteristics of 188 episodes occurring in tertiary care hospitals. Diagn Microbiol Infect Dis 58:77–82 [View Article][PubMed]
    [Google Scholar]
  15. Okawa Y., Miyauchi M., Kobayashi H. 2008; Comparison of pathogenicity of various Candida tropicalis strains. Biol Pharm Bull 31:1507–1510 [View Article][PubMed]
    [Google Scholar]
  16. Oksuz S., Sahin I., Yildirim M., Gulcan A., Yavuz T., Kaya D., Koc A. N. 2007; Phospholipase and proteinase activities in different Candida species isolated from anatomically distinct sites of healthy adults. Jpn J Infect Dis 60:280–283[PubMed]
    [Google Scholar]
  17. Pacheco M., Pisa D., García-Gómez P., Carrasco L., Juarranz Á. 2007; Attachment and entry of Candida famata in monocytes and epithelial cells. Microsc Res Tech 70:975–986 [View Article][PubMed]
    [Google Scholar]
  18. Pfaller M. A. 1992; Laboratory aids in the diagnosis of invasive candidiasis. Mycopathologia 120:65–72 [View Article][PubMed]
    [Google Scholar]
  19. Rüchel R., de Bernardis F., Ray T. L., Sullivan P. A., Cole G. T. 1992; Candida acid proteinases. J Med Vet Mycol 30:Suppl. 1123–132 [View Article][PubMed]
    [Google Scholar]
  20. Saegusa S., Totsuka M., Kaminogawa S., Hosoi T. 2007; Cytokine responses of intestinal epithelial-like Caco-2 cells to non-pathogenic and opportunistic pathogenic yeasts in the presence of butyric acid. Biosci Biotechnol Biochem 71:2428–2434 [View Article][PubMed]
    [Google Scholar]
  21. Silva S., Hooper S. J., Henriques M., Oliveira R., Azeredo J., Williams D. W. 2011; The role of secreted aspartyl proteinases in Candida tropicalis invasion and damage of oral mucosa. Clin Microbiol Infect 17:264–272 [View Article][PubMed]
    [Google Scholar]
  22. Sohn K., Senyürek I., Fertey J., Königsdorfer A., Joffroy C., Hauser N., Zelt G., Brunner H., Rupp S. 2006; An in vitro assay to study the transcriptional response during adherence of Candida albicans to different human epithelia. FEMS Yeast Res 6:1085–1093 [View Article][PubMed]
    [Google Scholar]
  23. Togni G., Sanglard D., Falchetto R., Monod M. 1991; Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis. FEBS Lett 286:181–185 [View Article][PubMed]
    [Google Scholar]
  24. Yang Y. L. 2003; Virulence factors of Candida species. J Microbiol Immunol Infect 36:223–228[PubMed]
    [Google Scholar]
  25. Zaugg C., Borg-Von Zepelin M., Reichard U., Sanglard D., Monod M. 2001; Secreted aspartic proteinase family of Candida tropicalis. Infect Immun 69:405–412 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.031195-0
Loading
/content/journal/jmm/10.1099/jmm.0.031195-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error