Yonsei Med J.  2006 Jun;47(3):354-358. 10.3349/ymj.2006.47.3.354.

Human Brain Astrocytes Mediate TRAIL-mediated Apoptosis after Treatment with IFN-gamma

• Lee J
• Shin JS
• Choi IH

• KMID: 1715853
• DOI: http://doi.org/10.3349/ymj.2006.47.3.354

Abstract

TNF-related apoptosis inducing ligand (TRAIL) expressions were studied in primary human brain astrocytes in response to pro-inflammatory cytokines. When astrocytes were treated with IL-1beta TNF-alphaor IFN-gamma TRAIL was induced in cultured fetal astrocytes. In particular, IFN-gammainduced the highest levels of TRAIL in cultured astrocytes. When astrocytes were pre-reated with IFN-gamma they induced apoptosis in TRAIL-sensitive Peer cells. Our results suggest that IFN-gamma modulates the expression of TRAIL in astrocytes, which may enhance cytotoxic sensitivity of infiltrating immune cells or brain cells other than astrocytes during inflammation of brain.

Keyword

IFN-gamma; astrocyte; TRAIL; apoptosis

MeSH Terms

Tumor Necrosis Factor-alpha/genetics/*metabolism
TNF-Related Apoptosis-Inducing Ligand
Membrane Glycoproteins/genetics/*metabolism
Interferon Type II/*pharmacology
Humans
Cells, Cultured
Astrocytes/*cytology/drug effects/metabolism
Apoptosis Regulatory Proteins/genetics/*metabolism
Apoptosis/*drug effects/physiology
Antineoplastic Agents/*pharmacology

Figure

• 

  Fig. 1 Effect of IFN-γ on TRAIL expression in astrocytes. 2 × 105 astrocytes were pre-treated with IFN-γ (250 U/mL) for 8 hr. (A) RPA was performed using 10 µg total RNA. IL-1 (100 U/mL) or TNF-α (750 U/mL) was treated for 8 h. (B) TRAIL expression was determined by semi-quantitative RT-PCR, and PCR product obtained was subjected to southern blotting. β-actin was used as an internal control. RT-PCR was performed in quadruplicate and representative results are shown. (C) For western blotting 40 µg of each cell homogenate were loaded on a 12% PAGE. Proteins were blotted onto a nitrocellulose membrane and incubated with 5 µg/mL of polyclonal mouse anti-human TRAIL antibody.

• 

  Fig. 2 Induction of apoptosis of Peer cells by astrocytes treated with IFN-γ. 2 × 105 fetal brain astrocytes were pre-treated with 250 U/mL of IFN-γ for 24 hr. Effector cells (astrocytes) and target cells (Peer cells) were cocultured for 24 hr at an E:T ratio of 1:1, 2:1 or 4:1. After collecting floating Peer cells, cell death was measured by staining with annexin V-FITC and propidium iodide. Total 1 × 105 cells were analyzed by a flow cytometry. Astrocytes not treated with IFN-γ were used as control effector cells. The assay was performed in quadruplicate and representative results are shown.

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