title: Analysis of inflammatory and lipid metabolic networks across RAW264.7 and thioglycolate-elicited macrophages.
abstract: Studies of macrophage biology have been significantly advanced by the availability of cell lines such as RAW264.7 cells. However, it is unclear how these cell lines differ from primary macrophages such as thioglycolate-elicited peritoneal macrophages (TGEMs). We used the inflammatory stimulus Kdo2-lipid A (KLA) to stimulate RAW264.7 and TGEM cells. Temporal changes of lipid and gene expression levels were concomitantly measured and a systems-level analysis was performed on the fold-change data. Here we present a comprehensive comparison between the two cell types. Upon KLA treatment, both RAW264.7 and TGEM cells show a strong inflammatory response. TGEM (primary) cells show a more rapid and intense inflammatory response relative to RAW264.7 cells. DNA levels (fold-change relative to control) are reduced in RAW264.7 cells, correlating with greater downregulation of cell cycle genes. The transcriptional response suggests that the cholesterol de novo synthesis increases considerably in RAW264.7 cells, but 25-hydroxycholesterol increases considerably in TGEM cells. Overall, while RAW264.7 cells behave similarly to TGEM cells in some ways and can be used as a good model for inflammation- and immune function-related kinetic studies, they behave differently than TGEM cells in other aspects of lipid metabolism and phenotypes used as models for various disorders such as atherosclerosis.
authors.affiliation: Department of Bioengineering, University of California at San Diego, La Jolla, CA 92093, USA.
authors.forename: Mano R
authors.initials: MR
authors.lastname: Maurya
authors.forename: Shakti
authors.initials: S
authors.lastname: Gupta
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Eoin
authors.initials: E
authors.lastname: Fahy
authors.forename: Ashok R
authors.initials: AR
authors.lastname: Dinasarapu
authors.forename: Manish
authors.initials: M
authors.lastname: Sud
authors.forename: H Alex
authors.initials: HA
authors.lastname: Brown
authors.forename: Christopher K
authors.initials: CK
authors.lastname: Glass
authors.forename: Robert C
authors.initials: RC
authors.lastname: Murphy
authors.forename: David W
authors.initials: DW
authors.lastname: Russell
authors.forename: Edward A
authors.initials: EA
authors.lastname: Dennis
authors.forename: Shankar
authors.initials: S
authors.lastname: Subramaniam
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D020869:Gene Expression Profiling; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D013864:Thioglycolates / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D016207:Cytokines; C506188:Kdo2-lipid A; D008070:Lipopolysaccharides; D013864:Thioglycolates
keywords: cell cycle; correlation analysis; inflammation; lipid and signaling networks; lipidomics; primary macrophage; toll-like receptor 4; transcriptomics