Plant expression of antigens of animal pathogens and its application for oral vaccination

动物病原体抗原的植物表达及其在口服疫苗中的应用

• 批准号: 12556053
• 负责人: SUGIMOTO Chihiro
• 金额: $ 8.45万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12556053/
• 关键词: Plant expression; transgenic plant; protozoa; tick; salivary gland; サイトカイン; 細胞成長因子; 粘膜免疫; ダニワクチン; ワクチン; 食べるワクチン; 細胞増殖因子; TGFβ

This project was aiming at expression of genes from animal pathogenic microbes, and development of mucosal vaccine as an injectable form into nasal or oral cavities in combination with cytokines. As model antigens, salivary antigens of ticks that transmit various protozoan parasite were investigated. Those antigens combined with cytokines such as interleukin-1 or -2 will be used as tick waccines.We introduced a bovine transforming growth factor beta gene into tobacco by using Agrobacterium system, and confirmed its expression in regenerated plant. Expression was enhanced by addition of legumine signal sequence. We also expressed bovine IL-1 and IL-2 in tobacco plants. Biological activities of these cytokines were confirmed by in vitro assays.Genes for tick cement proteins, serine proteinases, and their inhibitors (serpin) were cloned and their molecular structures were investigated. Biological activities of these molecules were also demonstrated. Vaccination of rabbit with recombinant cement proteins inhibited tick infestation, which suggested these proteins are good candidate for commercial production of tick vaccines.

该项目的目的是从动物致病性微生物中表达基因，并将粘膜疫苗作为一种与细胞因子结合使用的鼻或口腔注射形式。作为模型抗原，研究了传播各种原生动物寄生虫的滴答唾液抗原。这些抗原与细胞因子（例如白介素-1或-2）结合使用，将用作tick waccines。我们使用农业杆菌将牛转化的生长因子β基因引入烟草中，并确认其在再生植物中的表达。通过添加豆类信号序列来增强表达。我们还在烟草植物中表达了牛IL-1和IL-2。通过体外测定确认了这些细胞因子的生物学活性。tick胶结蛋白，丝氨酸蛋白酶及其抑制剂（SERPIN）的基因被克隆，并研究了它们的分子结构。还证明了这些分子的生物学活性。用重组水泥蛋白接种兔子疫苗抑制了壁虱侵染，这表明这些蛋白质是tick疫苗商业生产的良好候选者。

项目成果

Yokoyama N, Bork S.Nishisaka M, Hirata H, Matsuo T, Inoue N, Xuan X, Suzuki H, Sugimoto C, Igarashi I: "Roles of the Maltese Cross Form in the Development of Parasitemia and Protection against Babesia microti Infection in Mice"Infect Immun.. 71(1). 411-41

Yokoyama N、Bork S.Nishisaka M、Hirata H、Matsuo T、Inoue N、Xuan X、Suzuki H、Sugimoto C、Igarashi I：“马耳他十字形在小鼠寄生虫血症发展和预防田鼠巴贝斯虫感染中的作用

Sugimoto C., Ueda, S., Masuta, C., Ohya, K., Matsumura, T., Uyeda, I.: "Development of a new plant virus vector system and expression of human interferon alfa in broad beans")olecurar Farming (Toutant, J.P. & Balazs, E. ed; INRA Editions, France). 173-177

Sugimoto C.、Ueda, S.、Masuta, C.、Ohya, K.、Matsumura, T.、Uyeda, I.：“新型植物病毒载体系统的开发以及人干扰素α在蚕豆中的表达”）olecurar

T.Hataya, R.Arimoto, N.Suda, I.Uyeda: "Molecular characterization of Hop mosaic virus : its serological and molecular relationships to Hop latent virus"Arch Virol. 146. 135-1948 (2001)

T.Hataya、R.Arimoto、N.Suda、I.Uyeda：“啤酒花花叶病毒的分子特征：其与啤酒花潜伏病毒的血清学和分子关系”Arch Virol。

Ohya K., Itchoda N., Ohashi K., Onuma M., Sugimoto C., Matsumura T.: "Expression of biologically active human tumor necrosis factor-alpha in transgenic potato plant"Journal of Interferon & Cytokine Research. 22(3). 371-378 (2002)

Ohya K.、Itchoda N.、Ohashi K.、Onuma M.、Sugimoto C.、Matsumura T.：“转基因马铃薯植物中生物活性人类肿瘤坏死因子-α 的表达”干扰素杂志

Kakuda T., Sugimoto C., Onuma M.: "Epitope-mapping of antigen-specific T lymphocyte in cattle immunized with recombinant major piroplasm surface protein of Theileria sergenti"Journal of Veterinary Medical Science. 63(8). 895-901 (2001)

Kakuda T.、Sugimoto C.、Onuma M.：“用重组塞氏泰勒虫主要梨质表面蛋白免疫的牛中抗原特异性 T 淋巴细胞的表位作图”兽医医学科学杂志。