Time course study of cytokine mRNA expression in LPS-stimulated porcine alveolar macrophages.
The kinetics of cytokine mRNA expression was studied in porcine alveolar macrophages using an RT-PCR assay. The expression levels of IFN- gamma, IL-2, IL-4, IL-6, GM-CSF, IL-12 p35, and IL-12 p40 were examined after 2, 4, 14, 24, 48, and 72 h of incubation in unstimulated control and LPS-stimulated cells. The expression contents of IFN-gamma, IL-2, and IL-4 were not detected in both unstimulated and LPS-stimulated cells. On the other hand, the expression levels of IL-6, GM-CSF, and IL-12 in LPS-stimulated cells were almost always higher than those in control cells. Among those cytokines, IL-6 exhibited the predominant expression, and GM-CSF, IL-12 p40, and IL-12 p35 followed in the descending order. The times to reach the peak expression levels for IL-6, and GM-CSF, IL-12 p35, and IL-12 p40 were 14, and 24 h, respectively. After reaching the peak expression point, the expression levels of IL-6, GM-CSF, and IL-12 p40 reduced to the baseline by 72 h after stimulation, however, IL-12 p35 still kept a substantial expression by the same time. This study demonstrates that porcine alveolar macrophages primarily respond to express IL-6, GM-CSF, and IL-12 by LPS-stimulation and have a cytokine-specific expression profile during the stimulation time.
Choi
In-Soo
IS
Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Suwon 441-744, Korea.
Shin
Na-Ri
NR
Shin
Sung-Jae
SJ
Lee
Deog-Yong
DY
Cho
Young-Wook
YW
Yoo
Han-Sang
HS
D000818:Animals; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D007371:Interferon-gamma / Q000235:genetics; D007378:Interleukins / Q000235:genetics; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology*; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine / Q000235:genetics*
D016207:Cytokines; D007378:Interleukins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor