RAW 264-7 macrophage cell lines

Increased oxidative stress in the RAW 264.7 macrophage cell line is partially mediated via the S-nitrosothiol-induced inhibition of glutathione reductase (EC 1.6.4.2) (Butzer et al. 1999). Fujii et al. (2000) isolated a cDNA for rat glutathione reductase and constructed a baculovirus system to produce recombinant glutathione reductase on a large scale. NO donors (S-nitrosoglutathione, SIN-1, and S-nitroso-N-acetyl-D,L-penicillamine) inhibited the enzymatic activities of purified glutathione reductase. 

Nitric oxide synthases (EC 1.14.13.39), the inducible form of which has been obtained from the murine RAW 264.7 macrophage cell line (Hevel et al. 1991, Stuehr et al. 1991) was found by Liu et al. (1996) in the normal Lewis rat alveolar interstitium by immunostaining, whereas alveolar macrophages were iNOS negative. Interferon y maximally stimulated NO production by alveolar macrophages. 

In the first differential proteomics approach for the identification and characterization of proteins involved in both the inflammatory cascade and Leflunomide mode of action (Dax et al, 1998), the murine monocyte/macrophage cell line RAW 264.7 was employed and the RAW 264.7 proteomes of normal, unstimulated RAW 264.7 macrophages with the respective LPS-stimulated and/or drug-treated were compared. This cell line was chosen since RAW 264.7 cells have been used as a model system to study immune responses (Bahl et al, 1994) successfully before. 

Hussaini IM, LaMarre J, Lysiak JJ, et al. Transcriptional regulation of LDL receptor-related protein by IFN-y and the antagonistic activity of TGF-p(l) in the RAW 264.7 macrophage-like cell line. J Leukoc Biol 1996 59 733-9. 

Examples of cells lines in which this method has been validated are provided along with their growth media Raw 264.7 macrophages (culture in high glucose DMEM supplemented with 10% FBS and 2 mM GlutaMax ) PC-3 cells (culture in F-12K Medium supplemented with 10% FBS) Jurkat (culture in RPMl supplemented with 10% FBS) MDCK, L-cells, HeLa and COS-7 (culture in DMEM supplemented with 10% FBS). 

The murine macrophage cell line RAW 264.7 is routinely used to assess anti-inflammatory activity and NF-kB signaling in vitro. Inflammation can be induced in RAW 264.7 macrophages with lipopolysaccharides (EPS), a component found... 

Kobuchi H, Droy-Lefaix MT, Christen Y, Packer L. (1997). Ginkgo biloba extract (EGb 761) inhibitory effect on nitric oxide production in the macrophage cell line RAW 264.7. Biochem Pharmacol. 53(6) 897-903. 

Tumor necrosis factor inhibition. Ethanol (95%) extract of the rhizome, in cell culture at a concentration of 100 pg/mL, was inactive on macrophage cell line RAW 264.7 vs EPS induction of TNF-az° zp Tumor promotion inhibition. Ethyl acetate and methanol extracts of the dried rhizome, in cell culture at a concentration of 50 pg/mL, produced weak activity on G3H/ lOTl/2 cells vs tetradecanoyl phorbol acetate-induced acetate phospholipid synthesis. The hexane extract was inactiveZ . Ethanol (95%) and petroleum ether extracts of the dried rhizome, in cell culture at a concentration of 160 and 80 pg/mL, re-... 

Analysis by SDS-polyacrylamide gel electrophoresis of purified NOS from isolated HC from rats injected with killed Corynebacterium parvum (H), and from the murine macrophage cell line RAW 264-7 (M) (courtesy of D. Stuehr, The Cleveland Clinic, Cleveland, OH) which was exposed to LPS and IFNy. Crude cytosols were separated using ion exchange and affinity 2 5 -ADP Sepharose chromatography. Last step by gel filtration is equivalent to separation by molecular weight. 

Previous studies on the effects of N=0 production on the alloimmune response utilized bulk populations of responder and stimulator cells. In order to more clearly define the circumstances that induce -N=0 synthesis in allogeneic macrophage-lymphocyte cocultures, mouse splenocyte populations were depleted of accessory cells (>90% Thy 1.2+) and cultured with mitomycin-C-treated macrophage cell lines, as the alloantigen presenting cells. The P388D1 (H2 ) and RAW 264.7 (H2 ) macrophage lines were selected because... 

CTM = cytotoxic activity against mouse macrophage cell line (RAW 264.7) [167] ... 

Pycnogenol affects the activity of nitric oxide synthase and the production of nitric oxide in murine macrophages (RAW 264.7 cell line) activated by endotoxin (the lipopolysaccharide) and tumor necrosis factor (TNF)-... 

Channon, J. Y., and Leslie, C. C. 1990. A calcium-dependent mechanism for associating a soluble arachidonoyl-hydrolyzing phospholipase A2 with membrane in the macrophage cell line RAW 264.7. J. Biol Chem. 265 5409-5413. 

Membrane fractions of several cell lines derived from immunocompetent cells were previously analyzed by photoaffinity labeling (Williamson et al., 1995) and the key enzyme in pyrimidine biosynthesis, DHODH, was identified as one of the major targets of Leflimomide. For further elucidation of its mode of action, the cytosolic fraction of the monocyte/macrophage cell line RAW 264.7 was subjected to affinity chromatography to identify all cytosolic binding partners of Leflunomide. The cytosolic preparation was applied to an affinity column where the Leflunomide derivative A 95 0277 was coupled as a ligand (Figure 10.2). 

KOLODZIEJ, H., KAYSER, O., KIDERLEN, A. F., ITO, H., HATANO, T., YOSHIDA, T., FOO, L. Y., Proanthocyanidins and related compounds Antileishmanial activity and modulatory effects on nitric oxide and tumor necrosis factor-alpha-release in the murine macrophage-like cell line RAW 264.7, Biol. Pharm. 5m//., 2001,24, 1016-1021. 

Farber JM. A collection of mRNA species that are inducible in the RAW 264.7 mouse macrophage cell line by gamma interferon and other agents. Mol Cell Biol 1992 12 1535-1545. 

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