Limits...
Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination.

Hisahara S, Yuan J, Momoi T, Okano H, Miura M - J. Exp. Med. (2001)

Bottom Line: Here we show that caspase-11 plays crucial roles in OLG death and pathogenesis in EAE.Caspase-11 and activated caspase-3 were both expressed in OLGs in spinal cord EAE lesions.EAE susceptibility and cytokine concentrations in the CNS were significantly reduced in caspase-11-deficient mice.

View Article: PubMed Central - PubMed

Affiliation: Division of Neuroanatomy, Department of Neuroscience, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.

ABSTRACT
Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), are inflammatory diseases of the central nervous system (CNS) characterized by localized areas of demyelination. The mechanisms underlying oligodendrocyte (OLG) injury in MS and EAE remain unknown. Here we show that caspase-11 plays crucial roles in OLG death and pathogenesis in EAE. Caspase-11 and activated caspase-3 were both expressed in OLGs in spinal cord EAE lesions. OLGs from caspase-11-deficient mice were highly resistant to the cell death induced by cytotoxic cytokines. EAE susceptibility and cytokine concentrations in the CNS were significantly reduced in caspase-11-deficient mice. Our findings suggest that OLG death is mediated by a pathway that involves caspases-11 and -3 and leads to the demyelination observed in EAE.

Show MeSH

Related in: MedlinePlus

Caspase-11 plays an important role in cytokine-induced OLG cell death. (A) Immunoblot analysis of OLG lysates from wild-type control and caspase-11−/− mice using anti–caspase-1, -11, -8, and -3 antibodies. Even loading was demonstrated using an antitubulin antibody. Bar graph represents a densitometric analysis showing the relative amount of the processed caspase-3, produced by the treatment of cytokines between control and caspase-11 knockout mice, expressed as a percentage. (B) Primary cultured OLGs from caspase-11+/− and caspase-11−/− mice with or without the addition of 100 ng/ml IFN-γ. Many MBP+ OLGs from caspase-11−/− mice were observed 72 h after IFN-γ administration. Original magnification: 100×. (C) Percentages of cultured cells that represented live OLGs, calculated by counting MBP+ OLGs. TNF-α (200 ng/ml), IFN-γ (100 U/ml), and anti-Fas antibody (100 ng/ml) were administered to OLG cultures, which were fixed ∼72 h later. In each experiment, the OLGs collected from each independent well were counted, and the relative percentage of MBP+ cells was expressed as the mean ± SEM (error bars). Data were collected from three independent experiments. **P < 0.01 at each distance (Student's t test).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2195881&req=5

Figure 3: Caspase-11 plays an important role in cytokine-induced OLG cell death. (A) Immunoblot analysis of OLG lysates from wild-type control and caspase-11−/− mice using anti–caspase-1, -11, -8, and -3 antibodies. Even loading was demonstrated using an antitubulin antibody. Bar graph represents a densitometric analysis showing the relative amount of the processed caspase-3, produced by the treatment of cytokines between control and caspase-11 knockout mice, expressed as a percentage. (B) Primary cultured OLGs from caspase-11+/− and caspase-11−/− mice with or without the addition of 100 ng/ml IFN-γ. Many MBP+ OLGs from caspase-11−/− mice were observed 72 h after IFN-γ administration. Original magnification: 100×. (C) Percentages of cultured cells that represented live OLGs, calculated by counting MBP+ OLGs. TNF-α (200 ng/ml), IFN-γ (100 U/ml), and anti-Fas antibody (100 ng/ml) were administered to OLG cultures, which were fixed ∼72 h later. In each experiment, the OLGs collected from each independent well were counted, and the relative percentage of MBP+ cells was expressed as the mean ± SEM (error bars). Data were collected from three independent experiments. **P < 0.01 at each distance (Student's t test).

Mentions: To evaluate a possible role for caspase-11 in OLG cell death, we prepared purified OLGs as a primary culture from both wild-type and caspase-11−/− mice, and treated them with TNF-α or IFN-γ. After a 72-h incubation, we made cell lysates from the cultures and used them for immunoblotting with anti-caspase antibodies. In the lysates from wild-type OLGs, the expression of caspase-1, -11, and the processed form of caspase-3 was induced by TNF-α and IFN-γ (Fig. 3 A). In contrast, caspase-8 was not detected in either the cytokine-treated or the untreated lysates. These results were consistent with the immunohistochemical results shown in Fig. 1I and Fig. M. Interestingly, even though caspase-1 expression was induced by cytokines in the caspase-11−/− OLGs, the amount of processed caspase-3 was less than in wild-type OLGs. This result indicates that caspase-11 plays a more important role in activating caspase-3 than does caspase-1 in cytokine-induced OLG cell death. We next prepared purified OLGs from both caspase-11−/− and caspase-11+/− mice and treated them with TNF-α, IFN-γ, or the anti-Fas antibody Jo2. After a 72-h incubation, the cells were fixed and mature OLGs were identified by staining with an anti-MBP antibody to allow us to quantify the changes in the number of OLGs. Caspase-11–deficient OLGs were almost completely resistant to the cell death induced by IFN-γ and the anti-Fas antibody Jo2, and partially resistant to that induced by TNF-α (Fig. 3b and Fig. c). These results strongly suggest that caspase-11 is involved in promoting the OLG cell death induced by cytotoxic cytokines or Fas.


Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination.

Hisahara S, Yuan J, Momoi T, Okano H, Miura M - J. Exp. Med. (2001)

Caspase-11 plays an important role in cytokine-induced OLG cell death. (A) Immunoblot analysis of OLG lysates from wild-type control and caspase-11−/− mice using anti–caspase-1, -11, -8, and -3 antibodies. Even loading was demonstrated using an antitubulin antibody. Bar graph represents a densitometric analysis showing the relative amount of the processed caspase-3, produced by the treatment of cytokines between control and caspase-11 knockout mice, expressed as a percentage. (B) Primary cultured OLGs from caspase-11+/− and caspase-11−/− mice with or without the addition of 100 ng/ml IFN-γ. Many MBP+ OLGs from caspase-11−/− mice were observed 72 h after IFN-γ administration. Original magnification: 100×. (C) Percentages of cultured cells that represented live OLGs, calculated by counting MBP+ OLGs. TNF-α (200 ng/ml), IFN-γ (100 U/ml), and anti-Fas antibody (100 ng/ml) were administered to OLG cultures, which were fixed ∼72 h later. In each experiment, the OLGs collected from each independent well were counted, and the relative percentage of MBP+ cells was expressed as the mean ± SEM (error bars). Data were collected from three independent experiments. **P < 0.01 at each distance (Student's t test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195881&req=5

Figure 3: Caspase-11 plays an important role in cytokine-induced OLG cell death. (A) Immunoblot analysis of OLG lysates from wild-type control and caspase-11−/− mice using anti–caspase-1, -11, -8, and -3 antibodies. Even loading was demonstrated using an antitubulin antibody. Bar graph represents a densitometric analysis showing the relative amount of the processed caspase-3, produced by the treatment of cytokines between control and caspase-11 knockout mice, expressed as a percentage. (B) Primary cultured OLGs from caspase-11+/− and caspase-11−/− mice with or without the addition of 100 ng/ml IFN-γ. Many MBP+ OLGs from caspase-11−/− mice were observed 72 h after IFN-γ administration. Original magnification: 100×. (C) Percentages of cultured cells that represented live OLGs, calculated by counting MBP+ OLGs. TNF-α (200 ng/ml), IFN-γ (100 U/ml), and anti-Fas antibody (100 ng/ml) were administered to OLG cultures, which were fixed ∼72 h later. In each experiment, the OLGs collected from each independent well were counted, and the relative percentage of MBP+ cells was expressed as the mean ± SEM (error bars). Data were collected from three independent experiments. **P < 0.01 at each distance (Student's t test).
Mentions: To evaluate a possible role for caspase-11 in OLG cell death, we prepared purified OLGs as a primary culture from both wild-type and caspase-11−/− mice, and treated them with TNF-α or IFN-γ. After a 72-h incubation, we made cell lysates from the cultures and used them for immunoblotting with anti-caspase antibodies. In the lysates from wild-type OLGs, the expression of caspase-1, -11, and the processed form of caspase-3 was induced by TNF-α and IFN-γ (Fig. 3 A). In contrast, caspase-8 was not detected in either the cytokine-treated or the untreated lysates. These results were consistent with the immunohistochemical results shown in Fig. 1I and Fig. M. Interestingly, even though caspase-1 expression was induced by cytokines in the caspase-11−/− OLGs, the amount of processed caspase-3 was less than in wild-type OLGs. This result indicates that caspase-11 plays a more important role in activating caspase-3 than does caspase-1 in cytokine-induced OLG cell death. We next prepared purified OLGs from both caspase-11−/− and caspase-11+/− mice and treated them with TNF-α, IFN-γ, or the anti-Fas antibody Jo2. After a 72-h incubation, the cells were fixed and mature OLGs were identified by staining with an anti-MBP antibody to allow us to quantify the changes in the number of OLGs. Caspase-11–deficient OLGs were almost completely resistant to the cell death induced by IFN-γ and the anti-Fas antibody Jo2, and partially resistant to that induced by TNF-α (Fig. 3b and Fig. c). These results strongly suggest that caspase-11 is involved in promoting the OLG cell death induced by cytotoxic cytokines or Fas.

Bottom Line: Here we show that caspase-11 plays crucial roles in OLG death and pathogenesis in EAE.Caspase-11 and activated caspase-3 were both expressed in OLGs in spinal cord EAE lesions.EAE susceptibility and cytokine concentrations in the CNS were significantly reduced in caspase-11-deficient mice.

View Article: PubMed Central - PubMed

Affiliation: Division of Neuroanatomy, Department of Neuroscience, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.

ABSTRACT
Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), are inflammatory diseases of the central nervous system (CNS) characterized by localized areas of demyelination. The mechanisms underlying oligodendrocyte (OLG) injury in MS and EAE remain unknown. Here we show that caspase-11 plays crucial roles in OLG death and pathogenesis in EAE. Caspase-11 and activated caspase-3 were both expressed in OLGs in spinal cord EAE lesions. OLGs from caspase-11-deficient mice were highly resistant to the cell death induced by cytotoxic cytokines. EAE susceptibility and cytokine concentrations in the CNS were significantly reduced in caspase-11-deficient mice. Our findings suggest that OLG death is mediated by a pathway that involves caspases-11 and -3 and leads to the demyelination observed in EAE.

Show MeSH
Related in: MedlinePlus