Molecular and functional characterization of chicken IL-15

A cDNA encoding chicken interleukin-15 was cloned from a CD4+ T cell hybridoma expression library by screening with a rabbit antibody against a protein fraction of conditioned medium containing T cell growth promoting activity. The chicken IL-15 cDNA contains an open reading frame of 143 amino acids...

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Bibliographic Details
Published in:Developmental and comparative immunology 1999-03, Vol.23 (2), p.165-177
Main Authors: Choi, K D, Lillehoj, H S, Song, K D, Han, J Y
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Cattle
Chickens
CHO Cells
Cloning, Molecular
Cricetinae
DNA, Complementary
Humans
Interleukin-15 - genetics
Interleukin-15 - immunology
Molecular Sequence Data
Rabbits
Receptors, Antigen, T-Cell, gamma-delta - immunology
RNA, Messenger
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Tissue Distribution
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Summary:A cDNA encoding chicken interleukin-15 was cloned from a CD4+ T cell hybridoma expression library by screening with a rabbit antibody against a protein fraction of conditioned medium containing T cell growth promoting activity. The chicken IL-15 cDNA contains an open reading frame of 143 amino acids with a single potential N-linked glycosylation site. The predicted m.w. of the encoded protein (16 kDa) matched the size of an immunoreactive band on Western blots of E. coli expressing the recombinant IL-15. Amino acid and nucleotide sequence analyses of chicken IL-15 revealed 31% and 46% identity with bovine IL-15 respectively and lesser homologies to other mammalian IL-15s. Chicken IL-15 contained all 4 highly conserved cysteine residues present in mammalian IL-15 sequences. RT-PCR demonstrated that the chicken IL-15 gene is expressed in many tissues including spleen, intestine, and muscle and in established macrophage, T lymphoma and fibroblast cell lines. Activation of spleen cells with Con A enhanced the expression of IL-15 gene transcripts in a time-dependent manner. CHO-K1 cells transfected with the chicken IL-15 cDNA secreted a biologically active protein supporting the growth of Con A activated spleen lymphocytes. Continuous culture of spleen Con A lymphoblasts with chicken IL-15 over two months resulted in an enriched T lymphocyte population expressing the gammadeltaTCR, CD8alpha, and CD3 cell surface antigens.
ISSN:0145-305X
1879-0089
DOI:10.1016/S0145-305X(98)00046-9