Molecular cloning and expression of cynomolgus monkey interleukin-2 cDNA by the recombinant baculovirus system.
The cDNA encoding interleukin-2 (IL-2) of cynomolgus monkey was molecularly cloned from concanavalin-A-stimulated peripheral lymphocyte cDNA by the reverse transcription-polymerase chain reaction method. Cynomolgus monkey IL-2 cDNA displays a high degree of homology with the human equivalent both at nucleic acid (98%) and amino acid (96%) sequence levels, although having only one additional amino acid residue. Recombinant monkey IL-2 secreted by insect cells infected with the recombinant baculovirus migrated as two distinct bands with 15 and 18 kD corresponding to mature and precursor IL-2 molecules, respectively, which were revealed to cross-react with anti-human IL-2 polyclonal antibody in Western blot analysis. This monkey IL-2 exhibited high biological activity up to an order of 10(6) U/ml as assessed by an IL-2-dependent cell proliferation assay and was shown to be neutralized by anti-human IL-2 antibody.
Yabe
M
M
Department of Veterinary Science, National Institute of Health, Shinjuku-ku, Tokyo, Japan.
Matsuura
Y
Y
Tatsumi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D007313:Insecta; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism*; D007963:Leukocytes, Mononuclear; D008252:Macaca fascicularis / Q000235:genetics*; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D007376:Interleukin-2; D011994:Recombinant Proteins; D003208:Concanavalin A