Exposure of 16HBE cells to SC resulted in a statistically significant increase of hBD2 and hBD9 expression compared to that of the Selleck Androgen Receptor Antagonist untreated control cells or the cells exposed to the latex beads. The increase of defensin expression was also found in the cells exposed to RC and HF. However, this difference was significant only for hBD9
in the cells exposed to RC. The difference in expression of hBD2 by the cells exposed to RC and in the expression of hBD2 as well as hBD9 by the cells exposed to HF did not reach a significant level. There was no difference between defensin expression in the Selleckchem Tubastatin A untreated control cells and the cells exposed to the latex beads. Similar results were obtained with A549 cells. Figure 4 Analysis of mRNA levels for HBD2 and HBD9 in 16HBE cells exposed to A. fumigatus organisms. 16HBE cells (5 × 106) were grown in six well plates for 24 hours. The cells were then exposed to the different morphotypes of A. fumigatus or latex beads for 18 h. Cells were cultivated
learn more in a control well in the absence of A. fumigatus or the latex beads. Isolation of total RNA and synthesis of cDNA was performed as described in Methods. Specific primer pairs and the conditions of real time PCR are described in Table 2. The level of mRNA for defensins was measured in total RNA preparation by quantitative real time PCR as described in Methods. Expression of all genes was normalised to the expression of the endogenous reference gene GAPDH. The expression value in control cells
was used as the baseline. Data are calculated from three different experiments performed in triplicate. Means followed by the same letter are not significantly different. Neutralising anti-interleukine-1β antibody decreased defensin expression in cells exposed to swollen conidia Since A. fumigatus has been shown to induce IL-1β in airway epithelium, and since the analysis of kinetic of defensin expression showed that the Il-1β-induced response was faster than the one induced by fungi Decitabine molecular weight (Figure 3), we investigated whether or not observed A. fumigatus-induced defensin expression was related to Il-1 β synthesized during anti-fungal response. For this reason, neutralising anti-interleukine-1β antibody was added to the cells before exposure to A. fumigatus organisms. One of the defensins, hBD-9, was chosen for real time PCR analysis of the role of Il-1 β in defensin expression. The results of real time PCR revealed that relative gene expression was statistically significantly decreased in the cells treated with anti-Il-1 β antibody before exposure to SC, compared to the cells only exposed to SC (120 ± 5 versus 143 ± 10 respectively). Relative gene expression was also decreased in the cells treated with anti-Il-1 β antibody before exposure to RC or HF, but the difference did not reach a statistically significant level. The pre-treatment of the cells with normal mouse immunoglobulin before exposure to A.