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Characterization of lipopolysaccharide-stimulated cytokine expression in macrophages and monocytes
Haiqiu Huang1, Arnetta Fletcher, Yuge Niu
1Department of Nutrition and Food Science, University of Maryland, 0112 Skinner Building, College Park, MD 20742, USA.
Objective:
In vitro cell culture models are widely used in inflammation research; however, information regarding the time- and dose-dependency of inflammatory responses toward LPS in these cell lines is scattered in the literature.
Material:
J774A.1 mouse macrophage and THP-1 human monocyte cell lines.
Treatment:
J774A.1 and THP-1 cells were treated with 0-500 ng/mL lipopolysaccharide for 0-24 h.
Methods:
SRB and BCA tests were used to measure total protein. Real-time PCR was used to determine gene expression levels, and ELISA was used to assess the protein levels. One-way ANOVA and Tukey's Honestly Significant Difference test were used to test the significance levels.
Results:
In J774A.1 and THP-1 cells, cytokines responded in distinct patterns upon LPS stimulation in a time- and dose-dependent manner, and the differential regulation of the response to LPS between J774A.1 and THP-1 cells appeared to correlate with the differential regulation of TLR4 at the mRNA level.
Conclusion:
In summary, this study indicated that temporal and dose-dependent responses to LPS need to be controlled for and that extrapolation of data on mechanisms may differ between cell lines of different origin.
