Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs.
Abstract: cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid sequence of equine IL-1RII demonstrated 70, 60 and 58% similarity with human, mouse and rat sequences, respectively. Recombinant equine soluble IL-1RI and IL-1RII produced in insect cells bound recombinant equine IL-1alpha and IL-1beta. Furthermore, both receptors suppressed the growth inhibitory activities of equine IL-1alpha and IL-1beta toward A375 cells in a dose-dependent manner, indicating that the present equine IL-1RI and IL-1RII cDNA encodes biologically active proteins.
Publication Date: 2005-09-19 PubMed ID: 16176839DOI: 10.1016/j.vetimm.2005.08.018Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study explores the molecular properties and functional expressions of equine interleukin-1 type I and type II receptors. Derived from horse peripheral blood cells, these receptors display similarity with human, rat, and mouse sequences while indicating biological activity when interacting with other equine cells.
Explanation of the Research
In this research, scientists have cloned and analyzed two types of equine interleukin-1 receptors (IL-1RI and IL-1RII) from the peripheral blood cells of horses. These were generated based on the semi-conserved regions found in human and mouse IL-1RI and IL-1RII sequences.
- The study utilized cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells.
- Amplification and cloning of both type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) were executed using primers from the semi-conserved regions between human and mouse sequences.
Findings of the Study
The sequences of the equine receptors showcased significant similarity across various species.
- The amino acid sequence of equine IL-1RI showed 77% similarity with human sequences, 64% with mouse sequences, and 63% similarity with rat sequences.
- Conversely, the predicted amino acid sequence of equine IL-1RII demonstrated 70% similarity with human sequences, 60% with mouse sequences, and 58% with rat sequences.
Functional Expression of the Receptors
The recombinant form of these equine receptors produced in insect cells was capable of binding equine IL-1alpha and IL-1beta, indicating their biological activity.
- Type I and type II equine soluble interleukin-1 receptors bound recombinant equine interleukin-1 alpha and beta in insect cells.
- Both receptors suppressed the growth inhibitory activities of equine interleukin-1 alpha and beta towards A375 cells in a dose-dependent manner, showcasing they encode biologically active proteins.
Overall, these results suggest a functional role of these cloned equine interleukin-1 receptors, demonstrating their potential for further study in equine and comparative medicine.
Cite This Article
APA
Kirisawa R, Hashimoto N, Tazaki M, Yamanaka H, Ishii R, Hagiwara K, Iwai H.
(2005).
Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs.
Vet Immunol Immunopathol, 109(3-4), 219-231.
https://doi.org/10.1016/j.vetimm.2005.08.018 Publication
Researcher Affiliations
- Department of Veterinary Microbiology, School of Veterinary Medicine, Rakuno Gakuen University, Bunkyoudai-Midorimachi 582, Ebetsu, Hokkaido 069-8501, Japan. r-kirisa@rakuno.ac.jp
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Biological Assay
- Cloning, Molecular
- Horses / genetics
- Horses / immunology
- Horses / metabolism
- Interleukin-1 / immunology
- Molecular Sequence Data
- RNA / chemistry
- RNA / genetics
- Random Amplified Polymorphic DNA Technique / veterinary
- Receptors, Interleukin-1 / biosynthesis
- Receptors, Interleukin-1 / chemistry
- Receptors, Interleukin-1 / genetics
- Receptors, Interleukin-1 Type I
- Receptors, Interleukin-1 Type II
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Sequence Alignment
- Sequence Analysis, DNA
Citations
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