Nucleotide sequence analysis of viral genome related with pathogenesis of dengue hemorrhagic fever

登革出血热发病相关病毒基因组核苷酸序列分析

• 批准号: 07457078
• 负责人: IGARASHI Akira
• 金额: $ 3.65万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457078/
• 关键词: Dengue hemorrhagic fever; Viral genome; Nucleotide sequence; Amino acid sequence; Pathogenesis; デンダ出血熱; 3′非コード領域; 毒力; タイ国

Comparative nucleotide and deduced amino acid sequences were analyzed for 4 dengue type 2 virus strains isolated from patients with different clinical severities in Nakhon Phanom, Northeast Thailand in 1993, in order to obtain direct evidence for virulent virus theory of the pathogenesis of dengue hemorrhagic fever.The results showed followings : (1) Amino acid sequence of virus-specified proteins classified the isolates into 3 subtypes : subtype 1 was the isolate from the most severe DSS patient, subtype 2 were 2 isolates from DHF patients and 2 isolates from DF patients, and subtype 3 were 3 isolates from DF patients. (2) Secondary structure predicted for the 3'noncoding region (3'NCR) also classified the isolates into 3 subtypcs : subtype 1 was the isolate from DSS patient, subtype 2 were 2 isolates from DHF patients and 3 isolates from DF patients, and subtype 3 were 2 isolates from DF patients. These results indicated that the infection with virus strain possessing subtype 1 amino acid sequence and subtype 1 secondary structure of 3'NCR is expected to lead to severe clinical manifestation, while the infection with virus strains possessing subtype 3 amino acid sequence and subtype 3 secondary structure of 3'NCR is cxpected to be clinically mild. The plaque size formed on LLC-MK2 cells did not correlate with the structure of virus genome but rather with the serological response of the patients.Further studies were carried out on comparative sequence analysis on 4 strains of dengue type 4 virus isolated in Bangkok, Thailand, 1993. The result identified strain-specific amino acid replacements for the isolate from mild DF case as well as the isolate from relatively severe DHF grade II patients. These amino acid replacements were cxpected to influence the nature of the respective proteins. Effect of the observed molecular difference on the biological characteristics of virus isolates and relation to the pathogenesis of DHF remain to be investigated.

为获得登革出血热致病性强毒理论的直接证据，对1993年从泰国东北部帕诺姆分离的4株登革2型病毒进行了核苷酸和推导的氨基酸序列比较分析，结果表明：（1）病毒特异性蛋白的氨基酸序列将分离物分为3个亚型：亚型1为最严重DSS患者的分离株，亚型2为2株DHF患者的分离株和2株DF患者的分离株，3型为3株DF患者分离株。(2)根据3 '非编码区（3'NCR）的二级结构，将分离株分为3个亚型：1亚型为DSS患者分离株，2亚型为2株DHF患者分离株和3株DF患者分离株，3亚型为2株DF患者分离株。这些结果表明，具有1亚型氨基酸序列和1亚型二级结构的病毒株感染预期会导致严重的临床表现，而具有3亚型氨基酸序列和3亚型二级结构的病毒株感染预期会导致轻微的临床表现。本文对1993年泰国曼谷分离的4株登革4型病毒进行了序列比较分析。结果鉴定了来自轻度DF病例的分离株以及来自相对严重的DHF II级患者的分离株的菌株特异性氨基酸替换。预期这些氨基酸替换会影响相应蛋白质的性质。分子差异对病毒分离株生物学特性的影响以及与DHF发病机制的关系仍有待研究。

项目成果

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M.N.M.Mangada 等人：“从表现出不同疾病严重程度的患者中分离出的四种登革热 2 病毒的末端非编码区序列”病毒基因 14(1) (1996)。

五十嵐 章: "デング熱/デング出血熱" 化学療法の領域-シリーズ・目で見る感染症(45). 13. 4-5 (1997)

Akira Igarashi：“登革热/登革出血热”化疗领域系列视觉传染病（45）13. 4-5（1997）。

五十嵐 章: "東南アジアにおける日本脳炎とデング熱/デング出血熱" 長崎大学生涯学習教育センター運営委員会,長崎大学公開講座叢書9,アジアの時代を迎えて,II部, 212 (1997)

五十岚彰：“东南亚的日本脑炎和登革热/登革出血热”长崎大学终身学习教育中心指导委员会，长崎大学公开讲座系列 9，进入亚洲时代，第二部分，212（1997）

Igarashi A: Japanese encephalitis and dengue fever/dengue hemorrhagic fever (in Japanese). Publications of Public Lecture Nagasaki University, No.9, Facing the year of Asia, Vol II,Life-long Education Center, Nagasaki University, 123-137 (1997)

五十岚 A：日本脑炎和登革热/登革出血热（日语）。

五十嵐 章: "ウイルス性出血熱-ことにデング出血熱について-" 日本医学会,第108回日本医学会シンポジウム記録集,感染症, 98 (1997)

Akira Igarashi：“病毒性出血热 - 特别是关于登革出血热”日本医学会，第 108 届日本医学会研讨会记录，传染病，98（1997）