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Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment

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Astrocyte response to Junín virus infection.

Roberto G Pozner¹, Soledad Collado, Carolina Jaquenod de Giusti

¹Thrombosis 1 Laboratory, Haematological Research Institute, National Academy of Medicine, Pacheco de Melo 3081, 1425 Buenos Aires, Argentina.

Neuroscience Letters

|September 6, 2008

Summary

Junin virus (JV) infection in astrocytes increases nitric oxide (NO) production, which helps control viral replication and activate astrocytes. Inhibiting this response increases mortality, highlighting NO's protective role in the central nervous system.

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Area of Science:

Neuroscience
Immunology
Virology

Background:

Junin virus (JV) causes encephalitis, with previous studies showing increased inducible nitric oxide synthase (iNOS) expression and astrocyte reactions.
Inhibition of iNOS in prior studies led to higher mortality but reduced astrocytosis, suggesting a protective role for nitric oxide (NO).

Purpose of the Study:

To investigate if JV infection induces iNOS expression in cultured astrocytes.
To determine the relationship between JV infection, iNOS expression, viral replication, glial fibrillary acidic protein (GFAP) levels, and apoptosis.

Main Methods:

Cultured astrocytes were infected with JV.
iNOS and NO production were measured using immunocytochemistry, fluorescence-activated cell sorting (FACS), and nitrite/nitrate assays.
GFAP expression was assessed via immunocytochemistry, FACS, and RT-PCR.
The effects of iNOS inhibition on viral replication and GFAP expression were evaluated.

Main Results:

JV infection did not induce apoptosis in astrocytes.
JV infection increased iNOS and NO synthesis early in the infection.
iNOS inhibition abolished enhanced GFAP expression, indicating NO's direct involvement in astrocyte activation.
Inhibition of iNOS led to enhanced viral replication.

Conclusions:

Junin virus induces iNOS expression in astrocytes, leading to nitric oxide production.
Nitric oxide plays a dual role: reducing viral replication and enhancing astrocyte activation.
NO-mediated astrocyte activation is a beneficial response to JV-induced central nervous system damage.