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Truncated chicken interleukin-1beta with increased biologic activity.

Zsuzsa Gyorfy¹, Annette Ohnemus, Bernd Kaspers

¹Department of Virology, University of Freiburg, Germany.

Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research

|June 14, 2003

Summary

Chicken interleukin-1beta (ChIL-1beta) processing is inefficient but crucial for its activity. Cleavage significantly enhances ChIL-1beta

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Area of Science:

Immunology
Molecular Biology
Cytokine Research

Background:

Chicken interleukin-1beta (ChIL-1beta) is synthesized as a precursor.
Unlike mammalian counterparts, ChIL-1beta lacks a typical caspase-1 cleavage site, raising questions about its processing and activity modulation.

Purpose of the Study:

To investigate the proteolytic cleavage of ChIL-1beta.
To determine if cleavage modulates the biologic activity of ChIL-1beta.
To establish a bioassay for ChIL-1beta activity.

Main Methods:

Developed an avian indicator cell line with an NF-kappaB-regulated luciferase reporter gene for ChIL-1beta bioassay.
Utilized rabbit antiserum to identify NF-kappaB-stimulating activity in LPS-treated chicken macrophages.
Analyzed full-length and N-terminally truncated ChIL-1beta cDNAs for functional activity.

Main Results:

Established a sensitive and specific bioassay for ChIL-1beta.
Confirmed that NF-kappaB-stimulating activity in LPS-treated chicken macrophages is largely due to IL-1beta.
Demonstrated inefficient proteolytic processing of natural and recombinant ChIL-1beta.
Showed that both full-length and truncated ChIL-1beta cDNAs yield active cytokine.
A truncated ChIL-1beta molecule, resembling mature cytokine, exhibited over 100-fold enhanced biologic activity in mammalian cells.

Conclusions:

Proteolytic cleavage of ChIL-1beta is not highly efficient.
Precursor cleavage is critical for maximal ChIL-1beta biologic activity.
Truncated ChIL-1beta demonstrates significantly enhanced activity, highlighting the importance of processing for cytokine function.