Figure 1.

Epithelial cells (ECs) produce cytokines and chemokines upon CD40 ligation. (a) CD40 upregulation on vaginal ECs upon stimulation with 0, 5, 10, 20, 50, 100, or 200 IU ml⁻¹ IFNγ for 3 days. The height of the bars represents the CD40 mean fluorescence intensity as determined by flow cytometry. (b) Histogram of CD40 expression on vaginal ECs stimulated 3 days with 0 and 50 IU ml⁻¹ IFNγ. ELISA for IL8 (c) and RANTES (d) in cleared supernatants from IFNγ-pre-stimulated foreskin, vaginal, and cervical EC cultures (n=5–12) cocultured for 24 hours with control or CD40L-expressing L cells in the presence of IFNγ. *** Indicates P<0.0005. Reverse transcriptase-quantitative PCR of IL8 (e) and RANTES (f) expression by IFNγ-pre-stimulated vaginal ECs cocultured with L-control or L-CD40L cells in the presence of IFNγ for 0, 1, 2, 3, 6, 12, or 24 hours. Gene expression was normalized using GAPDH as the calibrator gene. Fold changes over 0 hours coculture were calculated and depicted. These data are representative for two to three independent experiments.