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Ornithine-containing lipids stimulate CD14-dependent TNF-alpha production from murine macrophage-like J774.1 and RAW
1Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan.
Abstract:
The ornithine-containing lipids (OL)-induced cytokine production pattern in macrophage-like J774.1 and RAW 264.7 cells was different from that in the peritoneal macrophages previously reported. OLs, as well as lipopolysaccharide (LPS) of Escherichia coli, strongly induced tumor necrosis factor (TNF) alpha but not interleukin (IL)-1beta in J774.1 cells. In the RAW cells, IL-1beta, TNF-alpha and prostaglandin E(2) were strongly induced by the OLs and LPS. OL- and serine-glycine-containing lipid (SGL)-induced TNF-alpha production in J774.1 and RAW 264.7 cells required serum. However, in CD14-deficient LR-9 cells, TNF-alpha was not induced by the OLs in the presence or absence of serum. OLs and a SGL almost completely inhibited the binding of (125)I-LPS to J774.1 cells. These results suggested that OLs and SGL activate macrophages via the CD14-dependent pathway.
Insights
Ornithine-containing lipids (OLs) and serine-glycine-containing lipids (SGLs) activate macrophages via a CD14-dependent pathway, influencing cytokine production like tumor necrosis factor (TNF)-alpha.
Area of Science:
- Immunology
- Cell Biology
- Lipid Biochemistry
Background:
- Macrophage activation is crucial for immune responses.
- Ornithine-containing lipids (OLs) are bacterial lipids with immunomodulatory potential.
- Previous studies showed varied macrophage responses to OLs.
Purpose of the Study:
- To investigate the cytokine production patterns induced by OLs in macrophage cell lines.
- To elucidate the role of CD14 in OL-mediated macrophage activation.
- To compare OL-induced responses with those induced by lipopolysaccharide (LPS).
Main Methods:
- Culturing of macrophage cell lines (J774.1, RAW 264.7, CD14-deficient LR-9).
- Measurement of cytokine (TNF-alpha, IL-1beta) and prostaglandin E(2) production.
- Assessment of (125)I-LPS binding to cells.
Main Results:
- OLs induced TNF-alpha but not IL-1beta in J774.1 cells, while inducing IL-1beta, TNF-alpha, and PGE(2) in RAW 264.7 cells.
- TNF-alpha induction by OLs and SGLs required serum in J774.1 and RAW 264.7 cells.
- OLs did not induce TNF-alpha in CD14-deficient LR-9 cells and inhibited LPS binding, suggesting CD14-dependent activation.
Conclusions:
- OLs and SGLs activate macrophages through a CD14-dependent mechanism.
- The cellular response to OLs varies between macrophage types.
- OLs and SGLs may act as antagonists to LPS binding via CD14.
