in vivo and in vitro study of asthma

To further development of our gene expression approach to biodosimetry, we have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to distinguish asthma and normal subjects. We analyzed gene expression patterns of the peripheral blood cells from asthma patients compared with those from normal subjects using microarray analyses. In addition, we analyzed gene expression patterns of the LPS or HDM-treated THP1 cells compared with those from non-treated THP1 cells. And we analyzed the microarray data through clustering analysis, signaling pathway analysis and others. Ten milliliters of peripheral blood of five mild asthma, five severe asthma and five normal subjects were centrifuged at 4000 rpm for 20 min to collect 0.4 mL of the buffy coat fraction for in vivo study. To induce an inflammatory response, for in vitro study, the THP1 cells were seeded in 12-well plates at a density of 1 × 106 cells/well in RPMI 1640 medium containing 0.5% fetal bovine serum and stimulated for 24 hrs with LPS or HDM extract. After stimulation the THP1 cells were harvested using a cell scraper and lysed for total RNA extraction. Total RNA was extracted from the buffy coat fractions and RNA was reverse transcribed into cDNA for microarray analysis