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Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia

Nature Protocols volume 1, pages 2767–2773 (2006)Cite this article

Abstract

This protocol describes the setup, maintenance and characteristics of models of epithelial Candida infections based on well-established three-dimensional organotypic tissues of human oral and vaginal mucosa. Infection experiments are highly reproducible and can be used for the direct analysis of pathogen–epithelial cell interactions. This allows detailed investigations of Candida albicans wild type or mutant strain interaction with epithelial tissue or the evaluation of the host immune response using histological, biochemical and molecular methods. As such, the models can be utilized as a tool to investigate cellular interactions or protein and gene expression that are not complicated by non-epithelial factors. To study the impact of innate immunity or the antifungal activity of natural and non-natural compounds, the mucosal infection models can be supplemented with immune cells, antimicrobial agents or probiotic bacteria. The model requires at least 3 days to be established and can be maintained thereafter for 2–4 days.

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Figure 1: Scanning electron micrograph of C. albicans interacting with the oral RHE.
Figure 2: Schematic diagram of the experimental design.
Figure 3: Light micrographs of oral RHE 24 h after inoculation with C. albicans SC5314 in the absence and presence of PMNs.

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Acknowledgements

We thank all the previous undergraduate and graduate students who have completed a project or a thesis in the laboratory and helped to build the foundations of these protocols. We thank Birgit Fehrenbacher, Renate Nordin, Helga Möller and Hannelore Bischof, University of Tuebingen, for excellent technical assistance and Gudrun Holland and Muhsin Özel, Robert Koch-Institute, for providing Figure 3. M.S. and G.W. were supported by the Deutsche Forschungsgemeinschaft (Sch 897/1; Sch 897/3), B.H. by the Robert Koch-Institute, the Deutsche Forschungsgemeinschaft (Hu 528/8; Hu 528/10) and the European Commission Union (QLK2-2000-00795; “Galar Fungail consortium”) and M.S. and J.N. by NIH grant R01 DE017514-01 and J.N. by a personal Wellcome Trust Value in People (VIP) award.

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Authors and Affiliations

  1. Department of Dermatology, Eberhard Karls University, Liebermeisterstr, 25, Tuebingen, D-72076, Germany

    Martin Schaller & Günther Weindl

  2. Robert Koch-Institute, FG16, Nordufer 20, Berlin, D-13353, Germany

    Katherina Zakikhany & Bernhard Hube

  3. Department of Oral Immunology, King's College London, King's College London Dental Institute, London, SE1 9RT, UK

    Julian R Naglik

  4. Friedrich Schiller University Jena and Department of Microbial Pathogenicity, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knoell Institute, Jena, 07745, Beutenbergstrasse 11a, Germany

    Bernhard Hube

Authors
  1. Martin Schaller
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  2. Katherina Zakikhany
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  3. Julian R Naglik
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  4. Günther Weindl
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  5. Bernhard Hube
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Correspondence to Martin Schaller.

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Schaller, M., Zakikhany, K., Naglik, J. et al. Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia. Nat Protoc 1, 2767–2773 (2006). https://doi.org/10.1038/nprot.2006.474

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