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Published on: July 11, 2015
Characterization of a cDNA encoding bovine interleukin 10: kinetics of expression in bovine lymphocytes
S M Hash1, W C Brown, A C Rice-Ficht
1Department of Veterinary Pathobiology, Texas A&M University, College Station 77843.
Insights
Researchers cloned and sequenced bovine interleukin 10 (IL10) cDNA from stimulated immune cells. The bovine IL10 sequence shows significant homology to human and mouse IL10, indicating conserved function in mammals.
Area of Science:
- Immunology
- Molecular Biology
- Comparative Genomics
Background:
- Interleukin 10 (IL10) is a critical cytokine involved in immune regulation.
- Understanding bovine IL10 is essential for comparative immunology and animal health research.
- Previous studies have characterized IL10 in various species, but bovine data was limited.
Purpose of the Study:
- To clone and sequence the complementary DNA (cDNA) encoding bovine Interleukin 10 (IL10).
- To analyze the sequence homology of bovine IL10 with its counterparts in other mammalian species.
- To investigate the expression pattern of bovine IL10 mRNA in immune cells.
Main Methods:
- Total cellular RNA was isolated from concanavalin A (ConA)-stimulated peripheral blood mononuclear cells (PBMC) of cattle.
- Complementary DNA (cDNA) was synthesized using reverse transcriptase and amplified via PCR with primers based on conserved regions of human and mouse IL10 genes.
- Northern blot analysis was performed to detect and quantify bovine IL10 mRNA transcripts.
Main Results:
- The nucleotide sequence of bovine IL10 cDNA exhibited 84%, 79%, and 78% homology to human, mouse, and rat IL10 cDNAs, respectively.
- The deduced amino acid sequence showed 77%, 71%, and 74% homology to human, mouse, and rat IL10 proteins.
- Northern blot analysis revealed a single 1.8-kb bovine IL10 mRNA transcript, with peak expression in ConA-stimulated T-cells between 8-24 hours, and minimal expression in lipopolysaccharide-activated macrophages.
Conclusions:
- The successful cloning and sequencing of bovine IL10 provide a valuable resource for immunological studies in cattle.
- The high degree of sequence homology suggests conserved functional roles of IL10 across mammalian species.
- Bovine IL10 is primarily expressed in activated T-cells, consistent with its known immunoregulatory functions.
Abstract:
A cDNA encoding bovine interleukin 10 (IL10) was cloned and sequenced using total cellular RNA derived from concanavalin A (ConA)-stimulated peripheral blood mononuclear cells (PBMC). A cDNA was produced with reverse transcriptase using oligo(dT) primers, and was amplified using primers chosen from consensus regions of the mouse and human IL10 genes. The nucleotide sequence derived from this cDNA shares 84, 79 and 78% homology with the human, mouse and rat cDNAs, respectively. The deduced amino-acid sequence shares an overall 77, 71 and 74% homology with the human, mouse and rat IL10 proteins, respectively. Northern blot analysis of the bovine IL10 mRNA reveals expression of a single 1.8-kb transcript, reaching maximal levels between 8 and 24 h, in ConA-stimulated peripheral T-cells, and weak expression in lipopolysaccharide-activated macrophages.
