Molecular analysis of intraerythrocytic parasite population dinamics and acquirement of biological diversity

红细胞内寄生虫种群动态的分子分析和生物多样性的获得

• 批准号: 10460126
• 负责人: SUGIMOTO Chihiro
• 金额: $ 8.32万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine (2000-2001)Hokkaido University (1998-1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10460126/
• 关键词: Theileria; Persistent infection; degenerative gel electrophoresisi; tick; salivary gland; antigenic wariation; 変成ゲル電気泳動法; 変異解析; rRNA遺伝子; 系統進化; EST; ゲノム解析; 遺伝子多型; DGGE; Theileria; ピロプラズマ; 原虫; 分子進化; リボソームRNA

During the course of Theileria orientalis infection in cattle, parasite population bearing different major piroplasm surface protein (MPSP) genes appear by turns, which may be one of immune evasion mechanisms of the parasite. Therefore, analysis of parasite population is important in understanding pathogenesis and developing control methods.In this study, degenerated gel electrophoresis methods for the analysis of parasite subpopulations at genetic level has been developed. By using this method, mutations within MPSP gene of C-type allele could be detected. From blood samples from cattle and buffalo, which were possibly infected with different Theileria, species was analyzed by rRNA gene-targeted PCR analysis and revealed mixed infection with 4 different species. The expressed sequence tag analysis of T. parva and EST mapping on chromosomes was carried out and 461 ESTs have been mapped.We further focused on tick saliva and salivary glands where tick, mammals and Theileria interact. Several proteins and their genes including cement proteins, proteinase inhibitors have been characterized. Probably, some of these tick proteins affect host protective responses and help parasite to invade into mammalian hosts.

在东方泰勒虫感染牛的过程中，携带不同MPSP基因的寄生虫种群交替出现，这可能是东方泰勒虫的免疫逃避机制之一。因此，寄生虫种群分析对于了解发病机制和制定控制方法具有重要意义。本研究开发了在遗传水平上分析寄生虫亚群的简并凝胶电泳方法。应用该方法可检测C型等位基因MPSP基因内的突变。从牛和布法罗的血液样品中，这可能是不同的泰勒虫感染，通过rRNA基因靶向PCR分析，发现混合感染4个不同的物种。表达序列标签分析表明，T.本文还对蜱、哺乳动物和泰勒虫相互作用的唾液和唾液腺进行了研究。一些蛋白质及其基因，包括水泥蛋白，蛋白酶抑制剂已被确定。这些蜱蛋白中的一些可能影响宿主的保护性反应，并帮助寄生虫侵入哺乳动物宿主。

项目成果

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Nishikawa, Y., Xuan, X., Nagasawa, H., Igarashi, I., Fujisaki, K., Otsuka, H., Mikami, T.: "Prevention of vertical transmission of Neospora caninum in BALB/c mice by recombinant vaccinia virus carrying NcSRS2 gene"Vaccine. 19. 1710-1716 (2001)

Nishikawa, Y.、Xuan, X.、Nagasawa, H.、Igarashi, I.、Fujisaki, K.、Otsuka, H.、Mikami, T.：“通过重组预防 BALB/c 小鼠中新孢子虫的垂直传播

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Nishikawa, Y., Inoue,N., Xuan, X., Nagasawa, H., Ikuo Igarashi, I., Fujisaki, K., Otsuka, H., Mikami, T.: "Protective efficacy of vaccination by recombinant vaccinia virus against Neospora caninum"Vaccine. 19. 1381-1390 (2001)

Nishikawa, Y.、Inoue,N.、Xuan, X.、Nagasawa, H.、Ikuo Igarashi, I.、Fujisaki, K.、Otsuka, H.、Mikami, T.：“重组痘苗病毒疫苗接种的保护功效

Mulenga, A, Sugino, M., Nakajima, M, Sugimoto, C. & Onuma, M: "Tick-encoded proteinase inhibitors (Serpins); potencial target antigens for tick vaccine development"J. Vet. Med. Sci.. 63. 1063-1069 (2001)

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