An immortalized cell line expresses properties of activated microglial cells

Bocchini, V.; Mazzolla, Rosanna; Barluzzi, Roberta; Blasi, Elisabetta; Sick, P.; Kettenmann, Helmut

doi:10.1002/jnr.490310405

Cited by 336 publications

(228 citation statements)

References 18 publications

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“…Stimulation of transformed murine microglia (BV-2 cells) (17) with purified, recombinant M-CSF induced cell migration; this was both dose-dependent and directional, as shown by checkerboard analysis, indicative of true chemotaxis (Fig. 3A).…”

Section: Resultsmentioning

confidence: 91%

Amyloid-β peptide–Receptor for Advanced Glycation Endproduct interaction elicits neuronal expression of macrophage-colony stimulating factor: A proinflammatory pathway in Alzheimer disease

Yan

Zhu²,

Fu³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

385

153

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In Alzheimer disease (AD), neurons are thought to be subjected to the deleterious cytotoxic effects of activated microglia. We demonstrate that binding of amyloidbeta peptide (A␤) to neuronal Receptor for Advanced Glycation Endproduct (RAGE), a cell surface receptor for A␤, induces macrophage-colony stimulating factor (M-CSF) by an oxidant sensitive, nuclear factor B-dependent pathway. AD brain shows increased neuronal expression of M-CSF in proximity to A␤ deposits, and in cerebrospinal fluid from AD patients there was Ϸ5-fold increased M-CSF antigen (P < 0.01), compared with age-matched controls. M-CSF released by A␤-stimulated neurons interacts with its cognate receptor, c-fms, on microglia, thereby triggering chemotaxis, cell proliferation, increased expression of the macrophage scavenger receptor and apolipoprotein E, and enhanced survival of microglia exposed to A␤, consistent with pathologic findings in AD. These data delineate an inflammatory pathway triggered by engagement of A␤ on neuronal RAGE. We suggest that M-CSF, thus generated, contributes to the pathogenesis of AD, and that M-CSF in cerebrospinal fluid might provide a means for monitoring neuronal perturbation at an early stage in AD.

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Section: Resultsmentioning

confidence: 91%

Amyloid-β peptide–Receptor for Advanced Glycation Endproduct interaction elicits neuronal expression of macrophage-colony stimulating factor: A proinflammatory pathway in Alzheimer disease

Yan

Zhu²,

Fu³

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

385

153

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“…BV-2 cells have been shown to avidly ingest 1-m fluorescent microspheres by phagocytosis (22,32). Surprisingly, uptake of microspheres was not enhanced by M-CSFR overexpression.…”

Section: Discussionmentioning

confidence: 99%

“…Transfections were carried out using mouse BV-2 and human SV-A3 microglial cells. The BV-2 immortalized microglial cell line has been characterized previously (21)(22)(23)(24). Because of phenotypic changes that occur at higher passages in BV-2 cells, 2 BV-2 cells used in these experiments were from passage 25 or lower.…”

Section: Microglial Cell Lines Plasmid Transfections and Tissue Culmentioning

confidence: 99%

Accelerated Phagocytosis of Amyloid-β by Mouse and Human Microglia Overexpressing the Macrophage Colony-stimulating Factor Receptor

Mitrasinovic

Murphy

2002

Journal of Biological Chemistry

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Alzheimer's disease (AD) 1 is characterized by amyloid-␤ peptide (A␤) plaques surrounded by microglia. A␤ is thought to be directly neurotoxic, and activated microglia are hypothesized to have negative effects on neurons through the release of effectors of inflammation (1). However, as brain macrophages microglia can clear A␤ by phagocytosis, primarily through macrophage scavenger receptors (MSR) (2-6). Immunization of transgenic mice modeling AD with A␤ results in clearance of plaques from the brain (7). Whereas some results suggest that microglial phagocytosis may be key in clearance of A␤ after immunization (8), other findings indicate that circulating antibodies may result in movement of A␤ out of the brain (9). This controversy has stimulated renewed interest in uptake of A␤ by microglia.A distinctive phenotypic feature of microglia surrounding A␤ plaques in APPV717F transgenic mice and in AD is enhanced expression of the macrophage colony-stimulating factor receptor (M-CSFR), translation product of the c-fms proto-oncogene (10, 11). Microglial M-CSFR expression is also increased after experimental ischemic and traumatic brain injury (12, 13). M-CSFR regulates proliferation, activation, and survival of cells in the monocyte-macrophage lineage through tyrosine kinase activation of diverse signal transduction pathways including: Src kinase, Ras-ERK, phosphoinositide 3-kinase, and p38 MAP kinase (14 -16). Deletion of M-CSFR expression results in decreased numbers of cells of the mononuclear phagocyte lineage (17). In AD macrophage colony-stimulating factor (M-CSF), the ligand for M-CSFR expressed by neurons and glia is also increased (18). Simultaneous increases in M-CSF and M-CSFR expression in the brain could result in significant changes in microglial function.We recently demonstrated that overexpression of M-CSFR by cultured microglia increases proliferation, stimulates release of pro-inflammatory and chemotactic cytokines, and induces a paracrine inflammatory response in a microglial-organotypic co-culture system (19). In the present study we sought to determine the effects of M-CSFR overexpression on A␤ phagocytosis by cultured mouse and human microglia. We hypothesized that M-CSFR-induced activation of microglia would increase their capacity to clear A␤ from culture medium. Although A␤ immunization clinical trials have been discontinued in humans for the present, identifying factors that enhance microglial clearance of A␤ may be of benefit in devising alternative means of decreasing A␤ burden in the brain in AD. EXPERIMENTAL PROCEDURES Microglial Cell Lines, Plasmid Transfections, and Tissue culture-The c-fms expression plasmid pTK1 was a gift from Dr. Rao Tekmal (Emory University, Atlanta, GA), and contains the complete mouse c-fms sequence under the control of an SV40 promoter (20). Transfections were carried out using mouse BV-2 and human SV-A3 microglial cells. The BV-2 immortalized microglial cell line has been characterized previously (21)(22)(23)(24). Because of phenotypic changes that occur at...

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“…Cell Culture-The murine BV-2 microglial cell line was obtained and characterized as described (20,42,43). The cells were cultivated in RPMI 1640 containing 10% heat-inactivated FBS (Hyclone Laboratories, Lagan, UK) supplemented with L-glutamine (4 mM), penicillin (100 units/ml), and streptomycin (0.1 mg/ml) in H 2 O-saturated 5% CO 2 atmosphere at 37°C.…”

Section: Methodsmentioning

confidence: 99%

S100B Protein Stimulates Microglia Migration via RAGE-dependent Up-regulation of Chemokine Expression and Release

Bianchi

Kastrisianaki

Giambanco

et al. 2011

Journal of Biological Chemistry

205

173

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The Ca 2؉ -binding protein of the EF-hand type, S100B, is abundantly expressed in and secreted by astrocytes, and release of S100B from damaged astrocytes occurs during the course of acute and chronic brain disorders. Thus, the concept has emerged that S100B might act an unconventional cytokine or a damage-associated molecular pattern protein playing a role in the pathophysiology of neurodegenerative disorders and inflammatory brain diseases. S100B proinflammatory effects require relatively high concentrations of the protein, whereas at physiological concentrations S100B exerts trophic effects on neurons. Most if not all of the extracellular (trophic and toxic) effects of S100B in the brain are mediated by the engagement of RAGE (receptor for advanced glycation end products). We show here that high S100B stimulates murine microglia migration in Boyden chambers via RAGE-dependent activation of Src kinase, Ras, PI3K, MEK/ERK1/2, RhoA/ROCK, Rac1/JNK/AP-1, Rac1/ NF-B, and, to a lesser extent, p38 MAPK. Recruitment of the adaptor protein, diaphanous-1, a member of the formin protein family, is also required for S100B/RAGE-induced migration of microglia. The S100B/RAGE-dependent activation of diaphanous-1/Rac1/JNK/AP-1, Ras/Rac1/NF-B and Src/Ras/PI3K/ RhoA/diaphanous-1 results in the up-regulation of expression of the chemokines, CCL3, CCL5, and CXCL12, whose release and activity are required for S100B to stimulate microglia migration. Lastly, RAGE engagement by S100B in microglia results in up-regulation of the chemokine receptors, CCR1 and CCR5. These results suggests that S100B might participate in the pathophysiology of brain inflammatory disorders via RAGEdependent regulation of several inflammation-related events including activation and migration of microglia. S100B is a Ca 2ϩ -binding protein of the EF-hand type abundantly expressed in astrocytes (1). S100B has been implicated in the regulation of astrocyte shape, migration, proliferation and differentiation via activation of the Src/PI3K module and PI3K-dependent stimulation of Akt and RhoA activities and hence of the supramolecular organization of F-actin (2). S100B also regulates the state of assembly of microtubules and type III intermediate filaments (3) and the cytosolic Ca 2ϩ concentration (4). In addition to having intracellular regulatory activities, S100B also exerts extracellular effects. Indeed, astrocytes secrete the protein constitutively and to a larger extent under the action of several stimuli including the proinflammatory cytokine, TNF-␣ (see for review Ref. 1). Moreover, levels of brain S100B are elevated in the aging brain and in several pathological conditions such as Alzheimer disease, brain infarct, epilepsy, and infectious diseases, as well as in Down syndrome (5, 6) in consequence of S100B human gene mapping to chromosome 21q22.3 (7). For example, hypertrophic astrocytes in peri-infarct areas and in neuritic plaques in Alzheimer disease and Down syndrome show elevated expression levels of S100B, and S100B can be detected outside hypert...

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An immortalized cell line expresses properties of activated microglial cells

Cited by 336 publications

References 18 publications

Amyloid-β peptide–Receptor for Advanced Glycation Endproduct interaction elicits neuronal expression of macrophage-colony stimulating factor: A proinflammatory pathway in Alzheimer disease

Amyloid-β peptide–Receptor for Advanced Glycation Endproduct interaction elicits neuronal expression of macrophage-colony stimulating factor: A proinflammatory pathway in Alzheimer disease

Accelerated Phagocytosis of Amyloid-β by Mouse and Human Microglia Overexpressing the Macrophage Colony-stimulating Factor Receptor

S100B Protein Stimulates Microglia Migration via RAGE-dependent Up-regulation of Chemokine Expression and Release

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