Immortalization of murine microglial cells by a v-raf / v-myc carrying retrovirus

Blasi, Elisabetta; Barluzzi, Roberta; Bocchini, V.; Mazzolla, Rosanna; Bistoni, Francesco

doi:10.1016/0165-5728(90)90073-v

Cited by 946 publications

(700 citation statements)

References 26 publications

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“…BV-2 cells, derived from an immortalized microglial cell line (Blasi et al, 1990) (kindly provided by Prof. K. Frei, University Hospital Zürich, Zürich, Switzerland), were maintained in the same microglial culture medium.…”

Section: Methodsmentioning

confidence: 99%

Suppression of Microglial Inflammatory Activity by Myelin Phagocytosis: Role of p47-PHOX-Mediated Generation of Reactive Oxygen Species

Liu¹,

Hao²,

Letièmbre³

et al. 2006

J. Neurosci.

111

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Multiple sclerosis (MS) is pathologically characterized by inflammatory demyelination and neuronal injury. Although phagocytosis of myelin debris by microglia and macrophages in acute MS lesions is well documented, its pathophysiological significance is unclear. Using real-time quantitative PCR, flow cytometry, ELISA, and reactive oxygen species (ROS) measurement assays, we demonstrated that phagocytosis of myelin modulates activation of microglial cells prestimulated by interferon-␥ (IFN-␥) or a combination of IFN-␥ and lipopolysaccharide with a biphasic temporal pattern, i.e., enhanced production of proinflammatory mediators during the first phase (Յ6 h), followed by suppression during the second (6 -24 h) phase. In this second phase, myelin phagocytosis leads to an enhanced release of prostaglandin E2 and ROS in microglia, whereas the production of anti-inflammatory cytokines (particularly interleukin-10) remains unchanged. Suppression of inflammatory microglial activation by myelin phagocytosis was reversed by treatment with superoxide dismutase and catalase, by inhibition of the NADPH-oxidase complex, or by specific knockdown of the NADPH-oxidase-required adaptor p47-phagocyte oxidase (PHOX). Furthermore, we observed that myelin phagocytosis destabilized tumor necrosis factor-␣ and interferon-induced protein-10 mRNA through an adenine-uridine-rich elements-involved mechanism, which was reversed by blocking the function of NADPH-oxidase complex. We conclude that phagocytosis of myelin suppresses microglial inflammatory activities via enhancement of p47-PHOX-mediated ROS generation. These results suggest that intervention in ROS generation could represent a novel therapeutic strategy to reduce neuroinflammation in MS.

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Section: Methodsmentioning

confidence: 99%

Suppression of Microglial Inflammatory Activity by Myelin Phagocytosis: Role of p47-PHOX-Mediated Generation of Reactive Oxygen Species

Liu¹,

Hao²,

Letièmbre³

et al. 2006

J. Neurosci.

111

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“…PM were purified by adherence to plastic in tissue culture clusters (Corning-Costar Italia, Milan, Italy) for 2 hours at 37°C in 5% CO 2 . The retrovirus immortalized, BV2 microglia cell line of Balb/c origin (Blasi et al, 1990), was kindly provided by Prof. Elisabetta Blasi, University of Modena, Italy. Cells were grown in RPMI 1640 (HyClone Laboratories, Logan, Utah) supplemented with 100 IU/ml of penicillin, 2 mM glutamine, 100 g/ml streptomycin, 20 mM Hepes buffer, and 5% FCS (HyClone).…”

Section: Cell Isolation and Treatmentmentioning

confidence: 99%

Macrophage Preconditioning with Synthetic Malaria Pigment Reduces Cytokine Production via Heme Iron-Dependent Oxidative Stress

Taramelli

Recalcati

Basilico

et al. 2000

Laboratory Investigation

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SUMMARY:Hemozoin (malaria pigment), a polymer of hematin (ferri-protoporphyrin IX) derived from hemoglobin ingested by intraerythrocytic plasmodia, modulates cytokine production by phagocytes. Mouse peritoneal macrophages (PM) fed with synthetic ␤-hematin (BH), structurally identical to native hemozoin, no longer produce tumor necrosis factor ␣ (TNF␣) and nitric oxide (NO) in response to lipopolysaccharide (LPS). Impairment of NO synthesis is due to inhibition of inducible nitric oxide synthase (iNOS) production. BH-mediated inhibition of PM functions cannot be ascribed to iron release from BH because neither prevention by iron chelators nor down-regulation of iron-regulatory protein activity was detected. Inhibition appears to be related to pigment-induced oxidative stress because (a) thiol compounds partially restored PM functions, (b) heme oxygenase (HO-1) and catalase mRNA levels were up-regulated, and (c) free radicals production increased in BH-treated cells. The antioxidant defenses of the cells determine the response to BH: microglia cells, which show a lower extent of induction of HO-1 and catalase mRNAs and lower accumulation of oxygen radicals, are less sensitive to the inhibitory effect of BH on cytokine production. Results indicate that BH is resistant to degradation by HO-1 and that heme-iron mediated oxidative stress may contribute to malaria-induced immunosuppression. This study may help correlate the different clinical manifestations of malaria, ranging from uncomplicated to severe disease, with dysregulation of phagocyte functions and promote better therapeutic strategies to counteract the effects of hemozoin accumulation. (Lab Invest 2000, 80:1781-1788.I ntraerythrocytic stage plasmodia digest hemoglobin in the food vacuole, where the globin is degraded and heme is detoxified, principally by polymerization/crystallization into hemozoin, an insoluble, microcrystalline derivative of ferri-protoporphyrin IX (␣ hematin, AH) (Bohle et al, 1997;Francis et al, 1997;Pagola et al, 2000;Slater et al, 1991). As parasites mature, hemozoin (malaria pigment) accumulates in the food vacuole and remains in the "residual body" after schizont rupture. Pigment is found inside host's phagocytes following ingestion of whole parasites or "residual bodies." Although a clinical correlation between the presence of pigment in circulating neutrophils and monocytes and disease severity is established (Nguyen et al, 1995), the nature of the interaction between pigment and phagocytes, as well as its contribution to the pathogenesis of malaria, and particularly its severe complications, are still controversial. Indeed, both pro-inflammatory (induction of tumor necrosis factor ␣ [TNF␣], interleukin-6 [IL-6], and chemotactic cytokines) (Pichyangkul et al, 1994;Prada et al, 1995;Sherry et al, 1995) and antiinflammatory activities (inhibition of respiratory burst and adhesion molecule expression and of TNF␣ and nitric oxide [NO] production) (Prada et al, 1995;Schwarzer et al, 1992;Taramelli et al, 1995Taramelli et al, , 1998 ha...

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“…The cells used were: proteose-peptone elicited peritoneal macrophages from CD1 mice; J774.1 macrophage cell line and BV2, a retrovirus transformed microglia cell line of C3H/HeJ origin [14]. Cells were maintained in CM and routinely split.…”

Section: Ceilsmentioning

confidence: 99%

Macrophage populations of different origins have distinct susceptibilities to lipid peroxidation induced by β‐haematin (malaria pigment)

et al. 1998

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We investigated the susceptibility of peritoneal mouse macrophages and macrophage and microglia cell lines to the peroxidative activity of [~-haematin, the synthetic polymer identical to native malaria pigment. The extent of lipid peroxidation, measured as production of thiobarbituric acid reactive substances (TBARS), was greater for peritoneal macrophages than for cell lines and microglia cells. TBARS production apparently was not attributable to the release of free iron from the protoporphyrin moiety, but related to lower glutathione content and different lipid composition of the cell membrane. These findings offer a new interpretation for the contentious immunomodulatory effects of [~-haematin reported for phagocytes of different origins.© 1998 Federation of European Biochemical Societies.Key words: Macrophage; [3-Hematin; Malaria pigment; Lipid peroxidation tathione [ll]. These findings suggest the involvement of oxidative mechanisms in the intracellular activity of [~-haematin, due to the known properties of haem-containing molecules as catalysts of lipid peroxidation [12]. This prompted us to verify whether (i) [3-haematin has prooxidant activity in macrophages and (ii) the ability of the different macrophage populations to cope with the [3-haematin-induced oxidative stress is related to a different membrane lipid composition and/or to the level of antioxidant defences. Materials and methods MiceSpecific pathogen-free female CD1 mice, 6-8 weeks old, were obtained from Charles River Italia (Calco, Como, Italy). Mice were maintained under conventional conditions and fed standard mouse pellet and water ad libitum.

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Immortalization of murine microglial cells by a v-raf / v-myc carrying retrovirus

Cited by 946 publications

References 26 publications

Suppression of Microglial Inflammatory Activity by Myelin Phagocytosis: Role of p47-PHOX-Mediated Generation of Reactive Oxygen Species

Suppression of Microglial Inflammatory Activity by Myelin Phagocytosis: Role of p47-PHOX-Mediated Generation of Reactive Oxygen Species

Macrophage Preconditioning with Synthetic Malaria Pigment Reduces Cytokine Production via Heme Iron-Dependent Oxidative Stress

Macrophage populations of different origins have distinct susceptibilities to lipid peroxidation induced by β‐haematin (malaria pigment)

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