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.

在牛的Theileria Orientalis感染过程中，寄生虫种群具有不同的主要雌性表面蛋白（MPSP）基因，这可能是寄生虫的免疫逃避机制之一。因此，对寄生虫种群的分析对于理解发病机理和开发控制方法很重要。在这项研究中，已经开发了在遗传水平下分析寄生虫亚群的退化的凝胶电泳方法。通过使用这种方法，可以检测到C型等位基因的MPSP基因内的突变。从牛和水牛的血液样本中，可能感染了不同的Theileria，通过以RRNA基因靶向PCR分析进行了分析，并揭示了4种不同种类的混合感染。对染色体上的T. parva和EST映射进行了表达的序列标签分析，并已绘制了461个EST。已经表征了几种蛋白质及其基因，包括水泥蛋白，蛋白酶抑制剂。这些壁虱蛋白中的一些可能会影响宿主保护性反应，并帮助寄生虫入侵哺乳动物宿主。

项目成果

Kawazu,S.: "Phylogenetic relationships of the benign Theileria species in cattle and Asian buffalo based on the major piroplasm surface protein (p33/34) gene sequences."Int.J.Parasitol.. 29・4. 613-618 (1999)

Kawazu, S.：“基于主要梨质表面蛋白 (p33/34) 基因序列的牛和亚洲水牛良性泰勒虫物种的系统发育关系。Int.J.Parasitol.. 613-618 (1999) ）

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

Chae,J.S.: "A study of the systematics of Theileria spp. based upon small-subunit ribosomal RNA gene sequences."Parasitol.Res.. 85・11. 877-883 (1999)

Chae, J.S.：“基于小亚基核糖体 RNA 基因序列的泰勒虫属系统学研究”。Parasitol.Res. 85・11 (1999)。

Mulenga,A.: "Molecular characterization of a Haemaphysalis longicornis tick salivary gland-associated 29-kilodaiton protein and its effect as a vaccine against tick infestation in rabbits."Infect.Immun.. 67・4. 1652-1658 (1999)

Mulenga, A.：“长角血蜱唾液腺相关 29-kilodaiton 蛋白的分子特征及其作为兔子蜱感染疫苗的作用。Infect.Immun.. 1652-1658 (1999)。

Sako,Y.: "Molecular cloning and characterisation of 23-kDa piroplasm surface proteins of Theileria sergenti and Theileria buffeli."Int.J.Parasitol.. 29・4. 593-599 (1999)

Sako, Y.：“塞氏泰勒虫和布氏泰勒虫的 23-kDa 梨质表面蛋白的分子克隆和表征。Int.J.Parasitol.. 593-599 (1999)”