Expression of Type II Nitric Oxide Synthase in Primary Human Astrocytes and Microglia: Role of IL-1β and IL-1 Receptor Antagonist

Judy Liu, Meng Liang Zhao, Celia F. Brosnan, Sunhee C. Lee

Research output: Contribution to journalArticle

148 Citations (Scopus)

Abstract

In this work, we studied the expression of type II nitric oxide synthase (NOS) in primary cultures of human astrocytes and microglia. Cytokine-activated human fetal astrocytes expressed a 4.5-kb type II NOS mRNA that was first evident at 8 h, steadily increased through 48 h, and persisted through 72 h. The inducing signals for astrocyte NOS II mRNA expression were in the order IL-1β + IFN-γ > IL-1β + TNF-α > IL-1β. SDS-PAGE analysis of cytokine-stimulated astrocyte cultures revealed an approximately 130-kDa single NOS II band that was expressed strongly at 48 and 72 h (72 h > 48 h). Specific NOS II immunoreactivity was detected in cytokine-treated astrocytes, both in the cytosol and in a discrete paranuclear region, which corresponded to Golgi-like membranes on immunoelectron microscopy. In human microglia, cytokines and LPS failed to induce NOS II expression, while the same stimuli readily induced TNF-α expression. In cytokine-treated human astrocytes, neither NOS II mRNA/protein expression nor nitrite production was inhibited by TGF-β, IL-4, or IL-10. In contrast, IL-1 receptor antagonist exerted near complete inhibition of NOS II mRNA and nitrite induction. Monocyte chemoattractant peptide-1 mRNA was induced in TGF-β-treated astrocytes, demonstrating the presence of receptors for TGF-β in astrocytes. These results confirm that in humans, cytokines stimulate astrocytes, but not microglia, to express NOS II belonging to the high output nitric oxide system similar to that found in rodent macrophages. They also show that the regulation of type II NOS expression in human glia differs significantly from that in rodent glia. A crucial role for the IL-1 pathway in the regulation of human astrocyte NOS II is shown, suggesting a potential role for IL-1 as a regulator of astrocyte activation in vivo.

Original languageEnglish (US)
Pages (from-to)3569-3576
Number of pages8
JournalJournal of Immunology
Volume157
Issue number8
StatePublished - Oct 15 1996

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Interleukin-1 Receptors
Microglia
Nitric Oxide Synthase Type II
Interleukin-1
Astrocytes
Cytokines
Messenger RNA
Nitrites
Neuroglia
Rodentia
Immunoelectron Microscopy
Chemotactic Factors
Interleukin-4
Interleukin-10
Cytosol
Polyacrylamide Gel Electrophoresis
Monocytes
Nitric Oxide
Macrophages

ASJC Scopus subject areas

  • Immunology

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Expression of Type II Nitric Oxide Synthase in Primary Human Astrocytes and Microglia : Role of IL-1β and IL-1 Receptor Antagonist. / Liu, Judy; Zhao, Meng Liang; Brosnan, Celia F.; Lee, Sunhee C.

In: Journal of Immunology, Vol. 157, No. 8, 15.10.1996, p. 3569-3576.

Research output: Contribution to journalArticle

Liu, Judy ; Zhao, Meng Liang ; Brosnan, Celia F. ; Lee, Sunhee C. / Expression of Type II Nitric Oxide Synthase in Primary Human Astrocytes and Microglia : Role of IL-1β and IL-1 Receptor Antagonist. In: Journal of Immunology. 1996 ; Vol. 157, No. 8. pp. 3569-3576.
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abstract = "In this work, we studied the expression of type II nitric oxide synthase (NOS) in primary cultures of human astrocytes and microglia. Cytokine-activated human fetal astrocytes expressed a 4.5-kb type II NOS mRNA that was first evident at 8 h, steadily increased through 48 h, and persisted through 72 h. The inducing signals for astrocyte NOS II mRNA expression were in the order IL-1β + IFN-γ > IL-1β + TNF-α > IL-1β. SDS-PAGE analysis of cytokine-stimulated astrocyte cultures revealed an approximately 130-kDa single NOS II band that was expressed strongly at 48 and 72 h (72 h > 48 h). Specific NOS II immunoreactivity was detected in cytokine-treated astrocytes, both in the cytosol and in a discrete paranuclear region, which corresponded to Golgi-like membranes on immunoelectron microscopy. In human microglia, cytokines and LPS failed to induce NOS II expression, while the same stimuli readily induced TNF-α expression. In cytokine-treated human astrocytes, neither NOS II mRNA/protein expression nor nitrite production was inhibited by TGF-β, IL-4, or IL-10. In contrast, IL-1 receptor antagonist exerted near complete inhibition of NOS II mRNA and nitrite induction. Monocyte chemoattractant peptide-1 mRNA was induced in TGF-β-treated astrocytes, demonstrating the presence of receptors for TGF-β in astrocytes. These results confirm that in humans, cytokines stimulate astrocytes, but not microglia, to express NOS II belonging to the high output nitric oxide system similar to that found in rodent macrophages. They also show that the regulation of type II NOS expression in human glia differs significantly from that in rodent glia. A crucial role for the IL-1 pathway in the regulation of human astrocyte NOS II is shown, suggesting a potential role for IL-1 as a regulator of astrocyte activation in vivo.",
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