Santos Renato L, Schoffelmeer Jeroen A, Tsolis Renée M, Gutiérrez-Pabello José A, Bäumler Andreas J, Adams L Garry
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University System Health Science Center, College Station 77843-4467, USA.
J Infect Dis. 2002 Aug 1;186(3):372-8. doi: 10.1086/341509. Epub 2002 Jul 11.
To identify host genes differentially expressed during Salmonella enterica serotype Typhimurium infection, an RNA differential display was made with total RNA extracted from ileal loops that were infected with Salmonella Typhimurium 2.5 h after infection. Down-regulated cDNA was identified in bovine Peyer's patches after infection that was highly homologous to a human plasma membrane calcium-transporting ATPase (PMCA). Differential expression of PMCA, evaluated by Northern analysis, was found to have more than a 4.6-fold decrease in expression of mRNA (size, approximately 5.1 kb). PMCA mRNA was detected by in situ hybridization exclusively within epithelial cells in the Peyer's patches. cDNA (4.4 kb) was amplified by rapid amplification of cDNA ends, cloned, and sequenced and showed a high homology to hPMCA. Bovine PMCA is down-regulated in epithelial cells of Peyer's patches after infection with Salmonella Typhimurium and, subsequently, may influence cellular calcium levels that contribute to the inflammatory processes in the pathogenesis of diarrhea.
为了鉴定在鼠伤寒沙门氏菌感染期间差异表达的宿主基因,利用感染鼠伤寒沙门氏菌2.5小时后从回肠袢提取的总RNA进行了RNA差异显示。感染后在牛派伊尔结中鉴定出下调的cDNA,其与人质膜钙转运ATP酶(PMCA)高度同源。通过Northern分析评估,发现PMCA的差异表达导致mRNA(大小约5.1 kb)表达下降超过4.6倍。通过原位杂交仅在派伊尔结的上皮细胞中检测到PMCA mRNA。通过cDNA末端快速扩增扩增出4.4 kb的cDNA,进行克隆和测序,结果显示其与hPMCA具有高度同源性。牛PMCA在感染鼠伤寒沙门氏菌后在派伊尔结的上皮细胞中表达下调,随后可能影响细胞钙水平,这在腹泻发病机制的炎症过程中起作用。