Cloning, expression, and tissue distribution of bovine interleukin-21.
Bovine interleukin-21 (IL-21) cDNA was cloned and sequenced from bovine peripheral blood lymphocytes (PBLs) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin (PHA), and 50 ng/ml phorbol 12-myristate 13-acetate (PMA) for 48 h. The open reading frame of the bovine IL-21 cDNA is 459 bp in length and encodes 152 amino acids. The predicted amino acid sequence is 78.2 and 58.5% homologous to the human and murine IL-21 amino acid sequences, respectively. Recombinant bovine IL-21 was expressed by a baculovirus expression system. The bovine IL-21 was processed to the mature form in insect cells and secreted to the supernatant confirmed by N-terminal amino acid sequencing. The recombinant bovine mature IL-21 induced the proliferation of human IL-2-dependent cells, ILT-MAT. The mRNA expression for bovine IL-21 was observed in the spleen, but not in the brain, heart, lung, liver, and kidney. The bovine IL-21 identified in this study may provide new methods for the enhancement of innate immunity in cows.
Muneta
Yoshihiro
Y
National Institute of Animal Health, 3-1-5 Kannondai Tsukuba, Ibaraki 305-0856, Japan. ymuneta@affrc.go.jp
Kikuma
Reiko
R
Yoshihara
Kazuhiro
K
Mori
Yasuyuki
Y
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002417:Cattle / Q000235:genetics* / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D007378:Interleukins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D000098546:Interleukin-21
D018076:DNA, Complementary; D007378:Interleukins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000098546:Interleukin-21