Identification of eterotoxin molecules in rotavirus NSP-4 and the application for the protection of diarrhea

轮状病毒NSP-4中肠毒素分子的鉴定及其在预防腹泻中的应用

• 批准号: 13460141
• 负责人: MINAMOTO Nobuyuki
• 金额: $ 9.66万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460141/
• 关键词: Rotavirus; NSP4; Diarrhea; Enterotoxin region; Suckling mice; Antigenic characterization; Nitric oxide; リアソータント; エンテロトキシン領域; 酸化窒素(NO)

The aim of this study was to determine the enterotoxin molecules in non-structural protein(NSP4) of rotaviruses which are known to be the main cause of serious diarrheal disease in infants and young mammalian animals, and to demonstrate the mechanisms underlying induction of diarrhea. In this study, group A avian rotaviruses were used. The results run as follows :1) An avian rotavirus, PO-13 strain was found to be infectious and have a similar level of virulence to that of the monkey rotavirus SA-11 in a heterologous suckling mouse model, but a turkey rotavirus, Ty-3, did not. (Virology, 2001)2)In spite of the induction of the diarrhea by virus, avian rotavirus NSP4s expressed in Escherichia coil acted as enterotoxins in suckling mice as did mammalian rotavirus NSP4s, despite great differences in the amino acid sequences. The enterotoxin molecules of PO-13 NSP4 existed in amino acid residues 109 to 135, a region similar to that reported in SA11 NSP4. (J.Virol., 2002)3)The homologies of the enterotoxin domains in NSP4s (50-59%) are higher than those (31-37%) of full-length NSP4s among the compared avian and mammalian rotaviruses. (Virus Res., 2002)4)It has demonstrated that the difference between degrees of virulence of rotaviruses for suckling mice is dependent on their outer capsid proteins, VP7 and VP4 by kinetics of reassortant strains in intestines of suckling mice. (Virology, 2003)5)From antigenic analysis of avian rotavirus NSP4 by monoclonal antibodies, it was showed that there are at least four major antigenic sites on the protein and that one of antigenic sites is widely conserved among a variety of rotaviruses. (Microbiol.Immunol., 2003)

本研究的目的是确定轮状病毒的非结构蛋白（NSP 4）中的肠毒素分子，并证明诱导腹泻的潜在机制。在本研究中，使用A组禽轮状病毒。结果表明：（1）在异种乳鼠模型中，禽轮状病毒PO-13株具有感染性，其毒力与猴轮状病毒SA-11相似，而火鸡轮状病毒Ty-3株则无感染性。（Virology，2001）2）尽管病毒诱导腹泻，但在大肠杆菌中表达的禽轮状病毒NSP 4s在乳小鼠中起肠毒素作用，与哺乳动物轮状病毒NSP 4s一样，尽管氨基酸序列存在很大差异。PO-13 NSP 4的肠毒素分子存在于氨基酸残基109至135处，该区域与SA 11 NSP 4中报道的区域相似。（J.Virol.，2002)3)在比较的禽类和哺乳动物轮状病毒中，NSP 4s的肠毒素结构域的同源性（50-59%）高于全长NSP 4s的同源性（31-37%）。（病毒研究，2002）4）通过对乳鼠肠道内轮状病毒株的动力学研究，证实了轮状病毒对乳鼠的毒力程度的差异取决于其外衣壳蛋白VP 7和VP 4。（Virology，2003）5）通过单克隆抗体对禽轮状病毒NSP 4的抗原性分析，表明该蛋白上至少有四个主要抗原位点，并且其中一个抗原位点在多种轮状病毒中广泛保守。（微生物免疫学，（2003年）

项目成果

Atoji, Y.: "Distribution of NADPH diaphorase-containing neurons in the pigeon central nervous system."Journal of Chemical Neuroanatomy. 21. 1-22 (2001)

Atoji, Y.：“鸽子中枢神经系统中含有 NADPH 黄酶的神经元的分布。”化学神经解剖学杂志。

Atoji, Y.: "Distribution of NADPH diaphorase-containing, neurons in the pigeon central nervous system"Journal of Chemical Neuroanatomy. 21. 1-22 (2001)

Atoji, Y.：“鸽子中枢神经系统中含有 NADPH 黄酶的神经元的分布”化学神经解剖学杂志。

Ito, H.: "Complete nucleotide sequence of group A avian rotavirus genome and a comparison with its counterparts of mammalian rotaviruses."Virus Research. 75(2). 123-138 (2001)

Ito, H.：“A 组禽轮状病毒基因组的完整核苷酸序列以及与其对应的哺乳动物轮状病毒的比较。”病毒研究。

Unno, T.: "Interaction between the M2-and M3-receptor subtypes in muscarinic electrical and mechanical responses of intestinal smooth muscles."Neurophysiology. 35. 290-301 (2003)

Unno, T.：“肠道平滑肌毒蕈碱电和机械反应中 M2 和 M3 受体亚型之间的相互作用。”神经生理学。

El-Mahmoudy, A., et al.: "Thymoquinone suppresses expression of inducible nitric oxide synthase in rat macrophages"International Immunopharmacology. 2. 1603-1611 (2002)

El-Mahmoudy, A. 等人：“百里醌抑制大鼠巨噬细胞中诱导型一氧化氮合酶的表达”国际免疫药理学。