反义寡聚核苷酸抗柯萨奇病毒感染的研究

孙红妹; 刘哲伟; 张霆

Abstract

孙红妹,刘哲伟,张霆.反义寡聚核苷酸抗柯萨奇病毒感染的研究[J].Chinese journal of Epidemiology,2000,21(4):295-297

反义寡聚核苷酸抗柯萨奇病毒感染的研究

Antisense oligonucleotides resistance to coxsackievirus B₃ infection in HeLa cells

Received:March 17, 2000

DOI：

KeyWord: 反义寡聚核苷酸柯萨奇病毒 HeLa细胞

English Key Word: Antisense oligodeoxynucleotides（ODNs） Coxsakievirus HeLa cells

FundProject:

Author Name	Affiliation
SUN Hongmei	Beijing Central Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China
LIU Zhewei	Beijing Central Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China
ZHANG Ting	Beijing Central Laboratory of Infection and Immunity, Capital Institute of Pediatrics, Beijing 100020, China

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Abstract:

目的针对柯萨奇病毒-3型（CVB3）基因组重要功能区设计系列反义寡聚核苷酸序列，通过实验观察特异性反义核酸对病毒感染细胞的抑制作用。方法通过病毒空斑形成试验及致细胞病变作用，了解反义核酸抑制及阻断CVB₃病毒感染细胞的能力。结果实验所设计的反义核酸中SCB₁对感染HeLa细胞的CVB₃病毒活性抑制作用较强，当病毒感染量为1MOI时，病毒活性抑制率高达90%；S_CB4和S_CB2均为75%；S_CB6有50%抑制作用；而S_CB3和S_CB5以及非特异性对照序列对CVB₃病毒基因基本没有任何作用。结论通过以上实验确定S_CB1、S_CB2、S_CB4等较好的抗病毒反义核酸序列，为进一步研究反义核酸抗病毒作用提供了重要的实验基础。

English Abstract:

Objective Antisense oligodeoxynucleotides （ODNs） hold great promise as therapeutic agents for the treatment of many human diseases. Oligonucleotides specifically hybrid with a selected sequence of RNA or DNA and inhibit the flow of genetic information DNA (or RNA) to proteins. In accordance with the present study, several oligonucleotide sequences that targeted _CVB3 RNA coding _Vp4, _Vp2, _Vp1, cleavage enzyme(P2A) or untranslational region were designed for observation. Methods Antiviral activities of these oligonucleotides were evaluated by plaque reduction assay and cytopathic effect (CPE). Cells were also treated with random oligonocleotides as a specificity control. Results Original ODNs, displaying sequence-specific inhibition ranging from 50% to 90% were identified. Three ODNs (at 5 microM concentration), which targeting at the pyrimidine - rich tract (S_CB1)， structural protein _Vp1 (S_CB4) and initiation of translation (S_CB2) were shown to exhibit inhibition of viral infection 75% to 90% at 1 MOI. There was only low antiviral activity in SCB5 and S_CB3, and random oligonucleotide were not found having antiviral activity. Oligonucleotides in this study were not toxic at 5 microM concentration. Conclusion A better antiviral antisense oligonucleotides as S_CB1 and S_CB4、S_CB2, could provide candidate antisense oligonucleotides for the treatment of _CVB3 infection.

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