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Species specificity of human interleukin-3 demonstrated by cloning and expression of the homologous rhesus monkey (Macaca mulatta) gene

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Author: Burger, H. · Leen, R.W. van · Dorssers, L.C.J. · Persoon, N.L.M. · Lemson, P.J. · Wagemaker, G.
Institution: Instituut voor Toegepaste Radiobiologie en Immunologie TNO
Source:Blood, 11, 76, 2229-2234
Identifier: 231098
Keywords: 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


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.