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.
Conclusion: 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.
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