J Korean Soc Microbiol.  1998 Dec;33(6):575-582.

Dose effect of LPS on Cytokine mRNA Expressions by Mouse Peritoneal Macrophages

• Kim SK
• Lee TY
• Kim HS
• Lee YS
• Lee MS
• Park HS

Abstract

Many studies for LPS-induced cytokines, chemokines gene expressions have been reported, but results are highly diverse. The dose of LPS used, cell type studied or other technical factors may contribute to the differences. We investigated the cytokine mRNA expressions, interleukin-2 (IL-2), IL-6, IL-10 and IFNr, on thioglycollate (TG) elicited-mouse peritoneal macrophages stimulated with various concentrations, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1 ug/ml and 10 ug/ml, of LPS used by reverse transcription polymerase chain reaction (RT-PCR) and northern blot analysis. TG- elicited peritoneal macrophages expressed IL-6 and IFN-r mRNA in a dose-dependent manner. The expressions of IL-2 and IL-10 mRNA were dose-independent manner. In northern blot analysis, IL-6 mRNA expression was detected only in 10 ug/ml LPS concentration for short stimulation time (0.5 h), while IL-10 mRNAs were expressed evenly in all LPS concentrations. The expression of IL-6 mRNA was sustained until 72 h at 10 ug/ml concentration only, but IL- 10 mRNAs of all cases of LPS concentrations were sustained evenly until then. These results suggest that the concentration of LPS required for cytokine induction must be determined differently from gene to gene types, and although LPS concentration 10 ug/ml has not been used as an ordinary concentration in vitro cytokine study, 10 ug/ml of LPS might be more appropriate concentration in the study of IL-6 expression on mouse peritoneal macrophages.

MeSH Terms

Animals
Blotting, Northern
Chemokines
Cytokines
Gene Expression
Interleukin-10
Interleukin-2
Interleukin-6
Macrophages, Peritoneal*
Mice*
Polymerase Chain Reaction
Reverse Transcription
RNA, Messenger*
Chemokines
Cytokines
Interleukin-10
Interleukin-2
Interleukin-6
RNA, Messenger