[On mechanism of how lipopolysaccharide induces dendritic cells to mature through TLRs signal pathway].
OBJECTIVE
To find out how lipopolysaccharide (LPS) induces dendritic cells (DC) to be more mature through Toll-like receptors (TLRs) signal pathway.


METHODS
Monocytes were isolated from heparinized whole blood. 7 days later the monocytes were induced into dendritic cells by adding rhGM-CSF and rhIL-4. Total cellular RNA of unstimulated and stimulated DC were extracted by LPS 24 h later. Semi-quantitative RT-PCR and agarose gel electrophoresis were conducted to analyze the PCR products. The expression of NF-kappaB was detected by laser scanning confocal microscopy (LSCM).


RESULTS
Expressions of TLR2, 3, 4 were significantly higher in monocytes-derived DC induced for 7 days, the plasm of DC was stained positive and the nucleus of DC was stained negative. After being stimulated by LPS for 24 h, the expressions of TLR2, 3, 4 decreased, and that of TLR4 was almost undetectable; the nucleus of DC was stained strong positive, the plasm of DC was stained positive.


CONCLUSIONS
The expressions of TLR2, 3, 4 on DC change significantly after being stimulated by LPS. And NF-kappaB shifts from the plasm to the nucleus. This signifies that LPS induces DC to be more mature in function.
Dong
Wei
W
Department of Immunology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Zhang
Bing
B
Zhu
Yue-ming
YM
Cai
Mei-ying
MY
Wei
Da-peng
DP
D000293:Adolescent; D000328:Adult; D002469:Cell Separation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology*; D005260:Female; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D009000:Monocytes / Q000166:cytology*; D016328:NF-kappa B / Q000096:biosynthesis; D015398:Signal Transduction; D051193:Toll-Like Receptors / Q000096:biosynthesis*
D008070:Lipopolysaccharides; D016328:NF-kappa B; D051193:Toll-Like Receptors