You are here
Characterization of caprine interleukin-4.
Title | Characterization of caprine interleukin-4. |
Publication Type | Journal Article |
Year of Publication | 2001 |
Authors | Snekvik, KR, Beyer, JC, Bertoni, G, Von Beust, BR, Baszler, TV, Palmer, GH, McElwain, TF, Cheevers, WP |
Journal | Vet Immunol Immunopathol |
Volume | 78 |
Issue | 3-4 |
Pagination | 219-29 |
Date Published | 2001 Feb 10 |
ISSN | 0165-2427 |
Keywords | Amino Acid Sequence, Animals, Antibodies, Monoclonal, Base Sequence, Blotting, Western, Cattle, Cloning, Molecular, COS Cells, DNA, Complementary, Goats, Interleukin-4, Mice, Molecular Sequence Data, Molecular Weight, Recombinant Proteins, Transfection |
Abstract | Caprine interleukin-4 (IL-4) cDNA was cloned from RNA of mitogen stimulated goat peripheral blood mononuclear cells utilizing reverse transcriptase-polymerase chain reaction. The sequence of caprine IL-4 cDNA corresponds to a 535 nucleotide mRNA with 5'- and 3'-untranslated regions and a 405 nucleotide open reading frame, the first 66 nucleotides of which encode a putative signal peptide. Mature IL-4 is a 12.8kDa protein containing six cysteine residues and two potential N-linked glycosylation sites and is highly homologous with other ruminant IL-4. The predicted molecular mass of mature unglycosylated IL-4 was confirmed by western blot of recombinant caprine IL-4 expressed in bacteria with a monoclonal antibody against a carboxyterminal peptide derived from the predicted amino acid sequence of bovine IL-4. Eukaryotic expression plasmids containing caprine IL-4 cDNA were used to characterize recombinant IL-4. Transcription of IL-4 mRNA was confirmed by transfection of COS-7 and goat synovial membrane cells, and recombinant IL-4 produced by stably transfected L929 cells inhibited inducible nitric oxide synthase in macrophages. Genetic immunization of mice with a caprine IL-4 cDNA expression plasmid induced antibodies against recombinant caprine IL-4 produced in bacteria. |
Alternate Journal | Vet. Immunol. Immunopathol. |
PubMed ID | 11292525 |
Grant List | K08-AI01553 / AI / NIAID NIH HHS / United States R01-AR43718 / AR / NIAMS NIH HHS / United States R21-AI42690 / AI / NIAID NIH HHS / United States |