Muneta Yoshihiro, Kikuma Reiko, Yoshihara Kazuhiro, Mori Yasuyuki
National Institute of Animal Health, 3-1-5 Kannondai Tsukuba, Ibaraki 305-0856, Japan.
Vet Immunol Immunopathol. 2003 Sep 15;95(1-2):73-80. doi: 10.1016/s0165-2427(03)00106-5.
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.
从用10微克/毫升刀豆球蛋白A（ConA）、10微克/毫升植物血凝素（PHA）和50纳克/毫升佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯（PMA）刺激48小时的牛外周血淋巴细胞（PBLs）中克隆并测序了牛白细胞介素-21（IL-21）cDNA。牛IL-21 cDNA的开放阅读框长度为459 bp，编码152个氨基酸。预测的氨基酸序列与人和小鼠IL-21氨基酸序列的同源性分别为78.2%和58.5%。重组牛IL-21通过杆状病毒表达系统表达。牛IL-21在昆虫细胞中加工成成熟形式，并通过N端氨基酸测序证实分泌到上清液中。重组牛成熟IL-21诱导人IL-2依赖细胞ILT-MAT的增殖。在脾脏中观察到牛IL-21的mRNA表达，但在脑、心脏、肺、肝脏和肾脏中未观察到。本研究中鉴定的牛IL-21可能为增强奶牛的先天免疫力提供新方法。
