Abstract
We established an in vitro adherence model with primarily cultured human keratinocytes as target cells which allows for the investigation of the molecular mechanisms that are responsible for Candida albicans host cell attachment in the initiation of cutaneous candidosis. The extent of C. albicans binding to cultured human keratinocytes was dependent on the yeast inoculum size and the incubation temperature. Heat and paraform-aldehyde treatment of yeasts completely abolished the binding activity of C. albicans. Of the different Candida species tested, C. albicans was by far the most adhesive species. C. albicans adherence was blocked by the acid protease inhibitor pepstatin A and the metabolic inhibitor sodium azide. The latter, however, was much less effective when yeasts were preincubated, suggesting that sodium azide was mainly acting on the keratinocytes. The extracellular matrix protein fibronectin was slightly inhibitory, whereas the fibronectin-derived peptides RGD and RGDS were not able to prevent attachment. PepTite-2000, another RGD-containing synthetic peptide, reduced C. albicans adherence by a margin of 25% (P < 0.005). CDPGYIGSR-NH2, which is a synthetic adhesive peptide derived from the laminin B chain, was much more efficient in its inhibitory activity than the RGD peptides and reduced C. albicans adherence to cultured human keratinocytes up to 76% (P < 0.001). Laminin itself and the synthetic pentapeptide YIGSR were less active. A dose-dependent reduction in adherence was also observed with collagen type III. Additionally, saccharides were tested for their potential to inhibit C. albicans attachment to keratinocytes. The most potent competitive saccharide inhibitors of C. albicans adherence to human keratinocytes were the amino sugars D-(+)-glucosamine and D-(+)-galactosamine with one isolate of C. albicans (4918) and D-(+)-glucosamine and alpha-D-(+)-fucose with another C. albicans isolate (Sp-1). Collectively, our data suggest the existence of multiple molecular mechanisms such as protein-protein, lectin-carbohydrate, and yeast-yeast coaggregational interactions that are responsible for optimal C. albicans attachment to cultured human keratinocytes.
Full text
PDF








Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Birkedal-Hansen H., Taylor R. E., Bhown A. S., Katz J., Lin H. Y., Wells B. R. Cleavage of bovine skin type III collagen by proteolytic enzymes. Relative resistance of the fibrillar form. J Biol Chem. 1985 Dec 25;260(30):16411–16417. [PubMed] [Google Scholar]
- Borg M., Rüchel R. Expression of extracellular acid proteinase by proteolytic Candida spp. during experimental infection of oral mucosa. Infect Immun. 1988 Mar;56(3):626–631. doi: 10.1128/iai.56.3.626-631.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bouchara J. P., Tronchin G., Annaix V., Robert R., Senet J. M. Laminin receptors on Candida albicans germ tubes. Infect Immun. 1990 Jan;58(1):48–54. doi: 10.1128/iai.58.1.48-54.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boukamp P., Petrussevska R. T., Breitkreutz D., Hornung J., Markham A., Fusenig N. E. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988 Mar;106(3):761–771. doi: 10.1083/jcb.106.3.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brassart D., Woltz A., Golliard M., Neeser J. R. In vitro inhibition of adhesion of Candida albicans clinical isolates to human buccal epithelial cells by Fuc alpha 1----2Gal beta-bearing complex carbohydrates. Infect Immun. 1991 May;59(5):1605–1613. doi: 10.1128/iai.59.5.1605-1613.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Calderone R. A. Host-parasite relationships in candidosis. Mycoses. 1989;32 (Suppl 2):12–17. doi: 10.1111/j.1439-0507.1989.tb02303.x. [DOI] [PubMed] [Google Scholar]
- Calderone R. A., Linehan L., Wadsworth E., Sandberg A. L. Identification of C3d receptors on Candida albicans. Infect Immun. 1988 Jan;56(1):252–258. doi: 10.1128/iai.56.1.252-258.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Detmar M., Orfanos C. E. Tumor necrosis factor-alpha inhibits cell proliferation and induces class II antigens and cell adhesion molecules in cultured normal human keratinocytes in vitro. Arch Dermatol Res. 1990;282(4):238–245. doi: 10.1007/BF00371643. [DOI] [PubMed] [Google Scholar]
- Dubertret L. Reconstruction of the human skin equivalent in vitro: a new tool for skin biology. Skin Pharmacol. 1990;3(2):144–148. doi: 10.1159/000210861. [DOI] [PubMed] [Google Scholar]
- Edwards J. E., Jr, Mayer C. L., Filler S. G., Wadsworth E., Calderone R. A. Cell extracts of Candida albicans block adherence of the organisms to endothelial cells. Infect Immun. 1992 Aug;60(8):3087–3091. doi: 10.1128/iai.60.8.3087-3091.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fukayama M., Calderone R. A. Adherence of cell surface mutants of Candida albicans to buccal epithelial cells and analyses of the cell surface proteins of the mutants. Infect Immun. 1991 Apr;59(4):1341–1345. doi: 10.1128/iai.59.4.1341-1345.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grant D. S., Tashiro K., Segui-Real B., Yamada Y., Martin G. R., Kleinman H. K. Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell. 1989 Sep 8;58(5):933–943. doi: 10.1016/0092-8674(89)90945-8. [DOI] [PubMed] [Google Scholar]
- Gustafson K. S., Vercellotti G. M., Bendel C. M., Hostetter M. K. Molecular mimicry in Candida albicans. Role of an integrin analogue in adhesion of the yeast to human endothelium. J Clin Invest. 1991 Jun;87(6):1896–1902. doi: 10.1172/JCI115214. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hasty D. L., Ofek I., Courtney H. S., Doyle R. J. Multiple adhesins of streptococci. Infect Immun. 1992 Jun;60(6):2147–2152. doi: 10.1128/iai.60.6.2147-2152.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayman E. G., Engvall E., Ruoslahti E. Concomitant loss of cell surface fibronectin and laminin from transformed rat kidney cells. J Cell Biol. 1981 Feb;88(2):352–357. doi: 10.1083/jcb.88.2.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hazen K. C., Brawner D. L., Riesselman M. H., Jutila M. A., Cutler J. E. Differential adherence of hydrophobic and hydrophilic Candida albicans yeast cells to mouse tissues. Infect Immun. 1991 Mar;59(3):907–912. doi: 10.1128/iai.59.3.907-912.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hazen K. C. Participation of yeast cell surface hydrophobicity in adherence of Candida albicans to human epithelial cells. Infect Immun. 1989 Jul;57(7):1894–1900. doi: 10.1128/iai.57.7.1894-1900.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heidenreich F., Dierich M. P. Candida albicans and Candida stellatoidea, in contrast to other Candida species, bind iC3b and C3d but not C3b. Infect Immun. 1985 Nov;50(2):598–600. doi: 10.1128/iai.50.2.598-600.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iwamoto Y., Robey F. A., Graf J., Sasaki M., Kleinman H. K., Yamada Y., Martin G. R. YIGSR, a synthetic laminin pentapeptide, inhibits experimental metastasis formation. Science. 1987 Nov 20;238(4830):1132–1134. doi: 10.1126/science.2961059. [DOI] [PubMed] [Google Scholar]
- Jimenez-Lucho V., Ginsburg V., Krivan H. C. Cryptococcus neoformans, Candida albicans, and other fungi bind specifically to the glycosphingolipid lactosylceramide (Gal beta 1-4Glc beta 1-1Cer), a possible adhesion receptor for yeasts. Infect Immun. 1990 Jul;58(7):2085–2090. doi: 10.1128/iai.58.7.2085-2090.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kennedy M. J. Adhesion and association mechanisms of Candida albicans. Curr Top Med Mycol. 1988;2:73–169. doi: 10.1007/978-1-4612-3730-3_4. [DOI] [PubMed] [Google Scholar]
- King R. D., Lee J. C., Morris A. L. Adherence of Candida albicans and other Candida species to mucosal epithelial cells. Infect Immun. 1980 Feb;27(2):667–674. doi: 10.1128/iai.27.2.667-674.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klotz S. A., Smith R. L. A fibronectin receptor on Candida albicans mediates adherence of the fungus to extracellular matrix. J Infect Dis. 1991 Mar;163(3):604–610. doi: 10.1093/infdis/163.3.604. [DOI] [PubMed] [Google Scholar]
- Korting H. C., Ollert M., Georgii A., Fröschl M. In vitro susceptibilities and biotypes of Candida albicans isolates from the oral cavities of patients infected with human immunodeficiency virus. J Clin Microbiol. 1988 Dec;26(12):2626–2631. doi: 10.1128/jcm.26.12.2626-2631.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee K. L., Buckley H. R., Campbell C. C. An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida Albicans. Sabouraudia. 1975 Jul;13(2):148–153. doi: 10.1080/00362177585190271. [DOI] [PubMed] [Google Scholar]
- Miyakawa Y., Kuribayashi T., Kagaya K., Suzuki M., Nakase T., Fukazawa Y. Role of specific determinants in mannan of Candida albicans serotype A in adherence to human buccal epithelial cells. Infect Immun. 1992 Jun;60(6):2493–2499. doi: 10.1128/iai.60.6.2493-2499.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ollert M. W., Calderone R. A. A monoclonal antibody that defines a surface antigen on Candida albicans hyphae cross-reacts with yeast cell protoplasts. Infect Immun. 1990 Mar;58(3):625–631. doi: 10.1128/iai.58.3.625-631.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ray T. L., Digre K. B., Payne C. D. Adherence of Candida species to human epidermal corneocytes and buccal mucosal cells: correlation with cutaneous pathogenicity. J Invest Dermatol. 1984 Jul;83(1):37–41. doi: 10.1111/1523-1747.ep12261661. [DOI] [PubMed] [Google Scholar]
- Ray T. L., Payne C. D. Scanning electron microscopy of epidermal adherence and cavitation in murine candidiasis: a role for Candida acid proteinase. Infect Immun. 1988 Aug;56(8):1942–1949. doi: 10.1128/iai.56.8.1942-1949.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ray T. L., Wuepper K. D. Experimental cutaneous candidiasis in rodents; II. Role of the stratum corneum barrier and serum complement as a mediator of a protective infalmmatory response. Arch Dermatol. 1978 Apr;114(4):539–543. doi: 10.1001/archderm.114.4.539. [DOI] [PubMed] [Google Scholar]
- Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
- Rotrosen D., Edwards J. E., Jr, Gibson T. R., Moore J. C., Cohen A. H., Green I. Adherence of Candida to cultured vascular endothelial cells: mechanisms of attachment and endothelial cell penetration. J Infect Dis. 1985 Dec;152(6):1264–1274. doi: 10.1093/infdis/152.6.1264. [DOI] [PubMed] [Google Scholar]
- Sandin R. L., Rogers A. L., Patterson R. J., Beneke E. S. Evidence for mannose-mediated adherence of Candida albicans to human buccal cells in vitro. Infect Immun. 1982 Jan;35(1):79–85. doi: 10.1128/iai.35.1.79-85.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sawyer R. T., Garner R. E., Hudson J. A. Arg-Gly-Asp (RGD) peptides alter hepatic killing of Candida albicans in the isolated perfused mouse liver model. Infect Immun. 1992 Jan;60(1):213–218. doi: 10.1128/iai.60.1.213-218.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schenkman S., Robbins E. S., Nussenzweig V. Attachment of Trypanosoma cruzi to mammalian cells requires parasite energy, and invasion can be independent of the target cell cytoskeleton. Infect Immun. 1991 Feb;59(2):645–654. doi: 10.1128/iai.59.2.645-654.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Segal E., Lehrer N., Ofek I. Adherence of Candida albicans to human vaginal epithelial cells: inhibition by amino sugars. Exp Cell Biol. 1982;50(1):13–17. doi: 10.1159/000163121. [DOI] [PubMed] [Google Scholar]
- Sobel J. D., Myers P. G., Kaye D., Levison M. E. Adherence of Candida albicans to human vaginal and buccal epithelial cells. J Infect Dis. 1981 Jan;143(1):76–82. doi: 10.1093/infdis/143.1.76. [DOI] [PubMed] [Google Scholar]
- Sohnle P. G., Frank M. M., Kirkpatrick C. H. Mechanisms involved in elimination of organisms from experimental cutaneous Candida albicans infections in guinea pigs. J Immunol. 1976 Aug;117(2):523–530. [PubMed] [Google Scholar]
- Srebrnik A., Segal E. Comparison of Candida albicans adherence to human corneocytes from various populations. Acta Derm Venereol. 1990;70(6):459–462. [PubMed] [Google Scholar]
- Tronchin G., Bouchara J. P., Robert R., Senet J. M. Adherence of Candida albicans germ tubes to plastic: ultrastructural and molecular studies of fibrillar adhesins. Infect Immun. 1988 Aug;56(8):1987–1993. doi: 10.1128/iai.56.8.1987-1993.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Varani J., Nickoloff B. J., Riser B. L., Mitra R. S., O'Rourke K., Dixit V. M. Thrombospondin-induced adhesion of human keratinocytes. J Clin Invest. 1988 May;81(5):1537–1544. doi: 10.1172/JCI113486. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Watt F. M., Green H. Stratification and terminal differentiation of cultured epidermal cells. Nature. 1982 Feb 4;295(5848):434–436. doi: 10.1038/295434a0. [DOI] [PubMed] [Google Scholar]
- el-Maghrabi E. A., Dixon D. M., Burnett J. W. Characterization of Candida albicans epidermolytic proteases and their role in yeast-cell adherence to keratinocytes. Clin Exp Dermatol. 1990 May;15(3):183–191. doi: 10.1111/j.1365-2230.1990.tb02069.x. [DOI] [PubMed] [Google Scholar]