Hyphal Invasion of Candida albicans Inhibits the Expression of Human β-Defensins in Experimental Oral Candidiasis  Qian Lu, Jayalath A.M.S. Jayatilake, Lakshman P. Samaranayake, Lijian Jin  Journal of Investigative Dermatology  Volume 126, Issue 9, Pages 2049-2056 (September 2006) DOI: 10.1038/sj.jid.5700346 Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Hyphal formation of C. albicans and immunohistochemical expression of human β-defensin-2 in reconstituted human oral epithelia (RHOE) after 12, 24, and 48hours of infection with C. albicans. PAS staining of C. albicans hyphae and immunohistochemical staining of hBD-2 were performed in each sample. (a–c) Hyphal invasion is observed in the RHOE infected with C. albicans SC5314 isolate, and (c) the hyphae penetrate the whole depth of RHOE after 48hours. (d–f) Hyphal formation instead of hyphal invasion is observed in the RHOE infected with C. albicans ATCC 90028. (g–i) No hyphal formation is observed in the RHOE infected by HLC52 or (j–l) HLC54 mutants. (m–o) hBD-2 is sparsely detected in the cytoplasm of the control RHOE. Intense intercellular expression of hBD-2 is detected in the RHOE infected with (a) C. albicans SC5314 and (d) ATCC isolates for 12hours, while (b and e) hBD-2 is undetectable at 24hours. (c) At 48hours, hBD-2 remains undetectable in the RHOE infected with C. albicans wild isolate but (f) it is upregulated in those infected with C. albicans ATCC isolate. (h) Upregulation of hBD-2 expression is also observed in the RHOE infected with C. albicans HLC52 mutant for 24hours (l) as well as those infected with HLC54 mutant for 48hours. (p) Normal goat IgG shows no specific staining in the RHOE. Scale bar=100μm. Journal of Investigative Dermatology 2006 126, 2049-2056DOI: (10.1038/sj.jid.5700346) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Immunohistochemical expression of human cytokeratin in RHOE after 12, 24, and 48hours of infection with C. albicans SC5314 isolate. (d–f) Cytokeratin is expressed in the cytoplasm of control RHOE. (a) It is expressed in the superficial layers of the RHOE infected with C. albicans SC5314 for 12hours. (b) At 24hours, intense expression of cytokeratin is observed, which tends to extend to deep layers. (c) At 48hours, spotty expression of cytokeratin is observed. Scale bar=100μm. Journal of Investigative Dermatology 2006 126, 2049-2056DOI: (10.1038/sj.jid.5700346) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 In situ hybridization expression of human β-defensin-2 mRNA in RHOE after 12, 24, and 48hours of infection with C. albicans. (m–o) Extensive expression of hBD-2 transcripts is detected in control RHOE (a) as well as in the RHOE infected for 12hours with C. albicans wild isolate, (d) ATCC 90028, and (g and j) the SAP mutants. At 24hours, hBD-2 mRNA is expressed only in a few cells near the basal layer of the RHOE infected by (b) C. albicans SC5314 and (e) ATCC isolates and (h) sap1–3 mutant. (k) No mRNA expression of hBD-2 is detected in the RHOE infected with sap4–6 mutant. At 48hours, no expression of hBD-2 mRNA occurs in the RHOE infected by (c) C. albicans SC5314 isolate or (i) sap1–3 and sap4–6 (l) mutants. (f) Whereas the RHOE infected with C. albicans ATCC isolate show positive expression of hBD-2 mRNA near the basal layer. (p) No specific signal is observed in the RHOE processed with hBD-2 sense probe. Scale bar=100μm. Journal of Investigative Dermatology 2006 126, 2049-2056DOI: (10.1038/sj.jid.5700346) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions