Title
Species specificity of human interleukin-3 demonstrated by cloning and expression of the homologous rhesus monkey (Macaca mulatta) gene
Author
Burger, H.
van Leen, R.W.
Dorssers, L.C.J.
Persoon, N.L.M.
Lemson, P.J.
Wagemaker, G.
Instituut voor Toegepaste Radiobiologie en Immunologie TNO
Publication year
1990
Abstract
To enable preclinical studies on homologous interleukin-3 (IL-3) in primate species, we isolated the gene encoding Rhesus monkey IL-3 (RhIL-3). The nucleotide sequence of the RhIL-3 gene displayed 92.9% homology with that of the human IL-3 (hIL-3) gene. The isolated RhIL-3 gene encodes a 143-amino acid (aa) precursor polypeptide, nine C-terminal residues shorter than the human protein. Protein homology was found to be 89.5% for the signal peptide (19 aa) and 80.5% for the mature protein (124 aa). Comparison of the human and RhIL-3 coding sequences showed that the majority of substitutions had occurred at amino acid replacement sites indicating a rapid evolution of the IL-3 protein. After expression of a genomic fragment in COS cells, RhIL-3 cDNA was constructed, which enabled large-scale production of the RhIL-3 polypeptide. RhIL-3 produced by Bacillus licheniformis and purified to homogeneity appeared to be approximately 100-fold more effective in stimulating Rhesus monkey hematopoietic progenitors than hIL-3, whereas RhIL-3 and hIL-3 showed comparable stimulatory activity on normal as well as malignant human hematopoietic cells. Thus, the rapid evolution of hIL-3 has resulted in a unidirectional species specificity, which most likely restricts the in vivo effects of hIL-3 in Macaca species.
Subject
Amino Acid Sequence
Animal
Bacillus
Base Sequence
Cell Line
Cloning, Molecular
Comparative Study
DNA
Exons
Gene Expression
Hematopoietic Stem Cells
Human
Interleukin-3
Introns
Macaca mulatta
Molecular Sequence Data
Sequence Homology, Nucleic Acid
Species Specificity
Transfection
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TNO identifier
231098
ISSN
0006-4971
Source
Blood, 76 (11), 2229-2234
Document type
article