Effect of halofuginone treatment on vaginal β defensin production.

<p>Mice, under pseudo-estrus conditions, were twice infected with 10⁷<i>Candida albicans</i> in vagina. Two days before and every two days after infection, mice were injected intraperitoneally with 5 µg/100 µl of halofuginone solution or diluent of halofuginone and were treated intravaginally with 10 pg of mouse rIL-17. At day 4, 8 and 14 after infection, mice were treated intravaginally with 10 µg of coelenterazine and imaged in the IVIS-200™ imaging system under anesthesia using 2.5% isoflurane. (A) Dot plot of total photon emission from the infected regions and dot plot of CFU in vaginal washes. The statistical analysis was performed using non-parametric Mann-Whitney U test. The median is indicated by a straight line. Data are representative of one out of two independent experiments with similar results. After 4, 8, 14, 21, 28 days from challenge, the vaginal lumen was washed with 150 µl of saline and vaginal cells were recovered for β-defensin analysis. (B) Mean of fluorescence MIF of β-defensin 1, β-defensin 2 and β-defensin 3 cells evaluated by FACS analysis. The vaginal cells recovered by vaginal washes were stained with rabbit anti-mouse BD1, goat anti-mouse BD2 or goat anti-mouse BD3 and goat anti-rabbit TRIC conjugate or rabbit anti-goat PE conjugate respectively. (C) Percentage of epithelial cells producing d β-defensin 2. Data are the mean±SD (n = 8 mice, 4 mice for each of two separate experiments). Cells of vaginal washes were stained with FITC anti mouse pan-cytokeratin and goat anti-mouse BD2 and rabbit anti-goat PE conjugate. Data are representative of one out of two independent experiments (total 8 mice). The statistical analysis was performed using Mann-Whitney U test. * <i>p</i><0.05, (infected halofuginone+IL-17 treated mice vs infected halofuginone treated mice).