Rabbit interleukin-1 receptor antagonist. Cloning, expression, functional characterization, and regulation during intestinal inflammation
| Autor: | F, Cominelli, M, Bortolami, T T, Pizarro, L, Monsacchi, M, Ferretti, M T, Brewer, S P, Eisenberg, R K, Ng |
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| Rok vydání: | 1994 |
| Předmět: |
Male
DNA Complementary Sialoglycoproteins Molecular Sequence Data Gene Expression Binding Competitive Polymerase Chain Reaction Cell Line Mice Escherichia coli Animals Humans Amino Acid Sequence Cloning Molecular DNA Primers Inflammation Base Sequence Sequence Homology Amino Acid Receptors Interleukin-1 Blotting Northern Colitis Recombinant Proteins Rats Interleukin 1 Receptor Antagonist Protein Kinetics Gene Expression Regulation RNA Rabbits Interleukin-1 |
| Zdroj: | The Journal of biological chemistry. 269(9) |
| ISSN: | 0021-9258 |
| Popis: | Genomic and cDNA clones for rabbit interleukin-1 receptor antagonist (IL-1ra) were isolate based on homology with the human, mouse, and rat IL-1ra gene. A partial genomic clone, obtained by screening a rabbit genomic library, contained coding sequences for the carboxyl-terminal 108 amino acids of rabbit IL-1ra. Two classes of cDNA for rabbit IL-1ra were obtained using RNA from inflamed rabbit colon tissue. One class of cDNA coded for a secreted form of IL-1ra, whereas the other coded for a putative intracellular form of rabbit IL-1ra. The latter form is similar to that isolated from human epithelial cells. A partially synthetic rabbit IL-1ra gene was constructed and expressed in Escherichia coli. The recombinant rabbit IL-1ra was purified to homogeneity by ion exchange chromatography. Its affinity was similar to that of human IL-1ra for the human and mouse type I IL-1 receptor. From the cDNA clone and the purified recombinant protein, specific probes were developed for measuring levels of rabbit IL-1ra mRNA and protein in normal and inflamed rabbit tissues. Unlike IL-1 alpha and IL-1 beta, IL-1RA mRNA and protein were present at detectable levels in normal rabbit colon. During the development of an experimental formalin-immune complex colitis, rabbit IL-1 alpha showed a dramatic increase in tissue levels, consistent with previous results; IL-1ra also increased 3-4-fold. Treatment of colitis rabbits with corticosteroids significantly suppressed neutrophil infiltration, corticosteroid treatment suppressed IL-1ra but not IL-1 alpha mRNA steady-state levels. Our observations demonstrate that IL-1 and IL-1ra synthesis is differentially regulated in healthy and inflamed intestinal tissue. |
| Databáze: | OpenAIRE |
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