Microarray analysis of differential expression of cell cycle and cell differentiation genes in cells infected with Lawsonia intracellularis.
Infection of intestinal crypt epithelial cells by the obligate intracellular bacterium Lawsonia intracellularis is directly linked to marked proliferation of the infected enterocytes within 3-5days post-infection. The virulence factor for this unique host cell-proliferative response is not known, but is considered to involve altered crypt cell cycle or differentiation events. McCoy mouse fibroblast cells were infected with L. intracellularis, and then harvested for expressed mRNA at daily time points, with matching non-infected control cell cultures. Mouse DNA microarray (>44,000 transcript targets) analysis of cDNA derived from matching mRNA samples showed over 40 identifiable genes with at least 4-fold changes between days 0 and 3 after infection with L. intracellularis. These included altered transcription of typical host cell 'alarm' response genes, such as interferon-related response genes Isgf3g and Igtp, known to be associated with invading microbial agents. Altered transcription of several genes in these cells known to be active in regulation of the cell cycle or cell differentiation genes, including usp18, Hr, Elavl2 and Slfn2, were also detected. The altered transcription of several of these genes via RT-PCR analysis was confirmed. The microarray-detected altered transcription of cell cycle and cell differentiation genes is of possible interest for links to Lawsonia-related disturbances in epithelial cell differentiation within the intestinal crypt, but this would need to be confirmed in intestinal epithelial cell studies.
Laboratory of Veterinary Immunology and Microbiology, College of Veterinary Medicine, Konkuk University, Seoul, South Korea.
Yu-Sik
YS
Oh
Joong-Bok
JB
Lee
Steven
S
McOrist
D000818:Animals; D002453:Cell Cycle / Q000235:genetics* / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics* / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology; D007413:Intestinal Mucosa / Q000166:cytology / Q000382:microbiology; D020639:Lawsonia Bacteria / Q000472:pathogenicity*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000662:veterinary*; D013552:Swine