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Abstract

Objectives

To clone equine interleukin 1α (IL-1α) and equine interleukin 1β (IL-1β) and determine their full-length cDNA sequences.

Procedure

The mRNA isolated from lipopolysaccharide-stimulated cultured equine monocytes was reverse transcribed, and a cDNA library was constructed in a λ phage. The cDNA library was screened by means of plaque hybridization with radiolabeled human IL-1α and IL-1ß cDNA probes. The cDNA nucleotide sequences for equine IL-1α and equine IL-1β were determined by use of the dideoxy chain termination technique. The cDNA sequences were analyzed, using computer software, for sequence characteristics and compared with sequences reported for other species.

Results

The cDNA for equine IL-1α was 1,728 base pairs in length with an ORF encoding a peptide of 270 amino acids with a predicted molecular mass of 30.823 kd. The cDNA for equine IL-1β was 1,473 base pairs in length with an ORF encoding a peptide of 268 amino acids with a predicted molecular mass of 30.342 kd. Similarity between amino acid sequence of equine IL- 1α and sequences for IL-1 α of other species ranged from 62.5 to 82.2%; similarity between amino acid sequence of equine IL-1ß and sequences for IL-1β of other species ranged from 62.5 to 66.4%. Similarity between amino acid sequences of equine IL-1α and equine IL-1β was 26%.

Conclusions and Clinical Relevance

Results establish a basis for studying the roles of interleukin 1 in healthy and diseased joints in horses. (Am J Vet Res 1998;59:704-711)

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To clone equine interleukin 1 receptor antagonist (IL-1 ra) and determine its full-length cDNA sequence.

Procedure

A cDNA library derived from lipopolysaccharide-stimulated equine monocytes was screened by means of plaque hybridization to radiolabeled equine IL-1ra DNA probes generated by means of the polymerase chain reaction. The cDNA nucleotide sequence for equine IL-1ra was determined by use of the dideoxy chain termination technique, analyzed by use of computer software for sequence characteristics, and compared with sequences reported for IL-1ra of other species.

Results

The cDNA for equine IL-1ra was 1,614 base pairs in length with an ORF encoding a peptide of 177 amino acids with a predicted molecular mass of 20.427 kd. Similarity between the amino acid sequence of equine IL-1ra and sequences for human, murine, rat, and lapine IL-1ra was 76%. Similarity between sequence for equine IL-1ra and sequences for equine interleukin-1α and equine interleukin-1ß were 22.6 and 24.6%, respectively.

Conclusion

Comparison of the sequence for equine IL-1ra with sequences for IL-1ra of other species indicated a high degree of conservation.

Clinical Relevance

Results establish a basis for studying the roles of interleukin-1 in healthy and diseased joints in horses. (Am J Vet Res 1998;59:712-716)

Free access
in American Journal of Veterinary Research