[Study on the inhibitory effect of antisense phosphorothioate oligodeoxynucleotide on coxsackie virus B replication in vitro].
OBJECTIVE
To investigate the antiviral effect of antisense phosphorothioate oligodeoxynucleotide (AODN) on CVB3 replication in vitro.


METHODS
In this study, 21 mer antisense phosphorothioate oligodeoxynucleotide which complemented to nt 581-601 in 5' NCR of Coxsackie Virus B3 (CVB3) RNA was used to investigate antiviral activity in Vero cells as a specific inhibitor of CVB3 replication. Specific AODN of CVB3 RNA, sense oligodeoxynucleotide (SODN) and randomized nonsense-sequence RODN were synthesized for effect comparison among them. Vero cells infected by CVB3 were transfected with different concentrations of AODN mediated by lipofectamine reagent, so that the cells could take in more AODN. Cell control and viral control were set up. The inhibitory effect of AODN on CVB3 replication was evaluated with a number of variables, including inhibitory rates of cytopathic effect (CPE), cell survival rates by MTT assay, inhibitory rates of CVB3 antigen by ELISA, inhibitory rates of RNA by dot blotting and 50% tissue culture infective dose (TCID50).


RESULTS
The specific AODN could significantly inhibit CPE of CVB3-infected Vero cells, decrease the production of antigen and RNA of CVB3 and viral titers, and increase cell survival rates, in a dose-dependent manner. The strongest inhibitory effect appeared at 48 hours after transfection, the most effective concentration of AODN was 10 micromol/L. On the other hand, 10 micromol/L SODN also showed weaker inhibitory effect on CPE of CVB3-infected cells, but no antiviral effect of 10 micromol/L RODN was shown. AODN showed no-inhibition on HSV-1 replication, but inhibited some enteroviruses such as CVB3, Polio-1 and Echo-6 to some extent.


CONCLUSIONS
These results indicate that nt 581-601 in 5' NCR of CVB3 RNA may play an important role in regulating CVB3 replication and AODN may have inhibitory effect on CVB3 replication.
Qi
X
X
Department of Epidemiology, Tianjin Medical University, Tianjin 300070, China.
Li
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X
Liu
M
M
D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis; D002522:Chlorocebus aethiops; D029822:Enterovirus B, Human / Q000187:drug effects* / Q000502:physiology; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D012367:RNA, Viral / Q000096:biosynthesis; D014709:Vero Cells; D014779:Virus Replication / Q000187:drug effects*
D000956:Antigens, Viral; D020319:Oligodeoxyribonucleotides, Antisense; D012367:RNA, Viral