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Saliva Promotes Candida albicans Adherence to Human Epithelial Cells

Department of Oral Sciences and Orthodontics, University of Otago, PO Box 647, Dunedin, New Zealand;

View larger version (21K): [in this window] [in a new window]   Figure 1. Adherence of C. albicans ATCC 10261 yeast cells to cultured monolayers of epithelial cell lines A549 (), HEp-2 (), or HET-1A () in the presence of increasing concentrations of pooled human whole saliva added to the assay. Numbers of C. albicans cells bound are expressed as percentages of the number of input cells (5.5 x 104 per well). Data are expressed as the means ± SE of quadruplicate determinations from three separate experiments. The mean adherence in the presence of saliva was significantly higher (P < 0.05) than that in the absence of saliva for all data points, except for the adherence to A549 cells when 20% saliva was added to the assay.

View larger version (51K): [in this window] [in a new window]   Figure 2. Adherence of C. albicans ATCC 10261 yeast cells to cultured monolayers of epithelial cell lines A549 (), HEp-2 (), or HET-1A () after pre-treatment of the yeast cells with increasing concentrations of pooled whole saliva. Yeast cells (2.2 x 106 mL-1) were pre-treated with whole saliva diluted in assay buffer and washed before use in adherence assays, at an input concentration of 1.1 x 106 mL-1. Results are expressed as the percentage increase in binding relative to the binding of yeast cells pre-treated with buffer alone, and are the means ± SE of quadruplicate determinations from three separate experiments. Values significantly (P < 0.05) greater than adherence of untreated yeast cells are marked with an asterisk (*). Values for adherence of yeast cells pre-treated with buffer alone were 17.1 ± 1.9%, 19.2 ± 2.1%, and 36.9 ± 4.2% for A549, HEp-2, and HET-1A, respectively.

View larger version (52K): [in this window] [in a new window]   Figure 3. Depletion of the adherence-promoting effect of whole saliva by pre-absorbed saliva samples (60% in AS buffer) with C. albicans ATCC 10261 yeast cells. Pooled saliva samples were pre-absorbed by incubation with various concentrations of yeast cells before use in adherence assays to cultured cell monolayers of epithelial cell lines A549 (), HEp-2 (), or HET-1A (). The yeast cells used to adsorb the saliva were removed by centrifugation, and the adherence assays performed with the absorbed saliva and a fresh preparation of yeast cells at an input concentration of 1.1 x 106/mL. Results are expressed as the percentage increase in binding relative to the adherence of the yeast cells in the absence of saliva added to the assay, and are the means ± SE of quadruplicate determinations from three separate experiments. Values significantly (P < 0.05) less than adherence of untreated yeast cells are marked with an asterisk (*).

View larger version (24K): [in this window] [in a new window]   Figure 4. Relationship between the amount of C. albicans-specific IgA in saliva samples and the adherence-promoting effect of saliva. The effect on C. albicans ATCC 10261 adherence to HEp-2 monolayers of 30 saliva samples taken from 11 individuals was determined. Values for yeast cell adherence were the means of quadruplicate determinations for each saliva sample, and expressed as percentages of the number of input cells (5.5 x 104 per well). Specific anti-C. albicans IgA in saliva samples was measured by ELISA titration, and results are expressed as the mean relative A492 values of triplicate determinations for 1:16 dilutions of saliva samples as described in MATERIALS & METHODS. Regression analysis of the data gave an r value of 0.68, P < 0.005.

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