Establishment of human microglial cell lines after transfection of primary cultures of embryonic microglial cells with the SV40 large T antigen

Janabi, Nazila; Peudenier, Sylviane; Héron, Bénédicte; Ng, Kim Heng; Tardieu, Marc

doi:10.1016/0304-3940(94)11792-h

Cited by 240 publications

(238 citation statements)

References 16 publications

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“…Astrocytes, 1321N1 (23), microglia cells, CHME3 (24), and neurons, SH-SY5Y (22) have been previously described. Human astrocytoma/ glioblastoma cell lines U87MG and U138MG were obtained from NIBSC (cat # ARP043 and ARP028, respectively).…”

Section: Methodsmentioning

confidence: 99%

The brain-specific factor FEZ1 is a determinant of neuronal susceptibility to HIV-1 infection

Haedicke

Brown

Naghavi

2009

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

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Neurons are one of the few cell types in the human body that do not support HIV type-1 (HIV-1) replication. Although the lack of key receptors is a major obstacle to infection, studies suggest that additional functions inhibit virus replication to explain the exquisite resistance of neurons to HIV-1. However, specific neuronal factors that may explain this resistance remain to be discovered. In a screen for antiviral factors using a fibroblast line chemically mutagenized and selected for resistance to retroviral infection, we recently identified induction of rat FEZ1 (fasciculation and elongation protein zeta-1), a brain-specific protein, as the cause of this resistance. When exogenously expressed in nonneuronal cell lines rat FEZ1 blocked nuclear entry of retroviral DNA. Here, we demonstrate that among human brain cells, neurons naturally express high levels of FEZ1 compared to astrocytes or microglia cells and are correspondingly less susceptible to infection with pseudotyped HIV-1 that bypasses receptor-mediated viral entry. Demonstrating that endogenous FEZ1 was functionally important in the resistance of neurons to HIV-1 infection, siRNA-mediated knockdown of endogenous FEZ1 increased the infectivity of neurons while sensitive brain cell types like microglia became more resistant upon FEZ1 overexpression. In addition, FEZ1 expression was not induced in response to IFN treatment. As such, in contrast to other widely expressed, IFN-inducible antiviral factors, FEZ1 appears to represent a unique neuron-specific determinant of cellular susceptibility to infection in a cell type that is naturally resistant to HIV-1.

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Section: Methodsmentioning

confidence: 99%

The brain-specific factor FEZ1 is a determinant of neuronal susceptibility to HIV-1 infection

Haedicke

Brown

Naghavi

2009

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

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“…The plasmid encoding herpes simplex virus thymidine kinase (HSV-TK) was obtained from Dr. L. A. Loeb (University of Washington). The CHME5 cell line, a human fetal microglia cell line transformed by SV40 large T antigen (26), was used for viral titering and was a gift from Dr. David Mock (University of Rochester). CHME5 cells were cultured in Dulbecco's modified Eagle's medium from CellGro containing 5% fetal bovine serum (HyClone).…”

Section: Methodsmentioning

confidence: 99%

Modification of Human Immunodeficiency Virus Type 1 Reverse Transcriptase to Target Cells with Elevated Cellular dNTP Concentrations

Jamburuthugoda

Chugh

Kim

2006

Journal of Biological Chemistry

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Section: Specific Domains Of A~selectively Mediate Human Microglia Ntmentioning

confidence: 99%

“…The human microglial cell line (clone 3), obtained by immortalization of embryonic brain cultures with a plasmid encoding for the large T antigen of SV4O, extensively characterized, was kindly provided by Dr. N. Janabi (Janabi et al, 1995) and was grown in Dulbecco's modified Eagle's medium containing 10% fetal calf serum at 37°Cin humidified 10% C0 2/90% air. For stimulation, cells were plated for RNA extraction in 10-cm-diameter dishes (Falcon; i0c ells per dish in 7 ml of William's medium E) and for protein analysis in six-well plates (Falcon; 2 X l0cells per well in 1.5 ml of William's medium E).…”

Section: Cell Culturementioning

confidence: 99%

“…Figure 6 shows that microghal NF-KB activation by A/3(25-35) several microglial antigens. Moreover, the cells could be activated to express class II major histocompatibility complex antigen upon interferon-y stimulation and to release cytokines after IL-i stimulation (Janabi et at., 1995).As a further typical feature of microghia, we found in line with previous studies in rats (Elkabes et al, 1996) that these human microglial cells also express the NTs of the NGF family (NGF, BDNF, NT-3, and NT-4/5), as well as the NGF receptor trkA and the low-affinity NT receptor p75.To examine the possibility that microglia synthesize and release trophic factors in addition to cytotoxins at sites of inflammation and injury, the effects of inflammatory cytokines, the C3a and different A/I peptides on microglial NT synthesis and secretion were measured.The cytokines IL-i/I and TNF-a synergistically activate microglial NGF expression via an NF-KB-sensitive mechanism, suggesting that these cytokines function as intercellular signals that orchestrate the responses of the CNS to injury (Griffin et al, 1989;Dickson et at., 1993). These cytokines, which are expressed within hours following injury, have been implicated in a wide range of pathological processes.…”

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confidence: 99%

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Inflammatory Signals Induce Neurotrophin Expression in Human Microglial Cells

Heese

Höck

Otten

1998

Journal of Neurochemistry

263

144

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Inflammatory processes involving reactive microglia, e.g., those associated with /1-amyloid containing neuritic and core plaques in Alzheimer's disease, appear to contribute to neuronal degeneration in the CNS. The fact that increased nerve growth factor (NGF) protein levels were found throughout brains of Alzheimer's disease patients led us to investigate neurotrophin synthesis in a human microglial cell line showing typical properties of human microglial cells, including expression of neurotrophins such as NGF, as well as the NGF receptor trkA and the low-affinity neurotrophin receptor p75. We found that the cytokines interleukin-1/3 and tumor necrosis factor-a synergistically stimulate microglial NGF transcription and protein release. Moreover, exposure of microglial cells to complement factor C3a induces NGF expression. To assess the role of the transcription factor nuclear factor-KB (NF-KB) in inflammatory mediator-induced microglial NGF expression, the effect of the NF-KB inhibitor pyrrolidine dithiocarbamate (PDTC) was analyzed. In the presence of PDTC, a dose-dependent inhibition of cytokine-activated NGF expression occurred. In contrast, the C3a-dependent stimulation of NGF synthesis was not influenced by PDTC. In addition, microglial neurotoxicitymediating /3-amyloid peptides A/1(1-40) and A~3(1-42) failed to alter NGF synthesis, whereas A/9(25-35) specifically induced NF-KB-dependent microglial NGF expression. In conclusion, inflammatory signals (cytokines and complement factors), as well as A/3(25-35), are potent stimulators of human microglial NGF synthesis involving NF-KB-dependent and -independent mechanisms. Microglial secretion of neurotrophins appears to be involved in early processes of neuronal regeneration. Key Words: Nerve growth factor-Cytokines-Complement factors-Alzheimer's disease. J. Neurochem. 70, 699-707 (1998).ing free oxygen intermediates, proteases, excitatory amino acids, arachidonic acid derivatives, and cytokines, and have been implicated in the destruction of neurons in trauma (Banati et al., 1993;Thanos et al., 1993), Alzheimer's disease (McGeer et al., 1987(McGeer et al., , 1993 Giulian et al., 1995Giulian et al., , 1996McGeer and McGeer, 1995;Kaltschmidt et al., 1997), and human immunodeficiency virus infection (Giulian et al., 1990;Dickson et al., 1993). Moreover, in vitro studies have revealed that microglia/brain macrophages can secrete growth factors [e.g., basic fibroblast growth factor and nerve growth factor (NGF) 1~known to enhance survival and fiber outgrowth of different types of CNS neurons Mallat et al., 1989;Shimojo et al., 1991;Araujo and Cotman, 1992;Elkabes et al., 1996). For their survival and maintenance of function, several classes of neurons rely on the continuous supply of neurotrophic factors, including NGF. After injury, increased synthesis of NGF occurs in glial cells and in target tissues (Thoenen and Barde, 1980;Levi-Montalcini and Calissano, 1986; Levi-Montalcmi, 1987). In response to CNS injury, both microglia and astrocytes undergo str...

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Establishment of human microglial cell lines after transfection of primary cultures of embryonic microglial cells with the SV40 large T antigen

Cited by 240 publications

References 16 publications

The brain-specific factor FEZ1 is a determinant of neuronal susceptibility to HIV-1 infection

The brain-specific factor FEZ1 is a determinant of neuronal susceptibility to HIV-1 infection

Modification of Human Immunodeficiency Virus Type 1 Reverse Transcriptase to Target Cells with Elevated Cellular dNTP Concentrations

Inflammatory Signals Induce Neurotrophin Expression in Human Microglial Cells

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