Study on inhibition of persistent infection of Coxsackievirus B group by antisense phosphorothioate oligodeoxynucleotide in vitro

Objective: To study the inhibition of CVB3 and CVB5 in persistent infection by 21 mer antisense phosphorothioate oligodeoxynucleotide (AODN) in vitro. Methods:AODN was synthesized and wrapped by Lipofectamine regeant. This AODN was transfected with different concentration to CVB3-ECV304 and CVB5-ECV304 cell models. The inhibitory effects of AODN were evaluated with a number of indexes, including inhibition of CPE on Vero cells caused by supernanant of cell models, inhibitory rates of CVB3 and CVB5 antigens by ELISA, viral RNA expression inside the cell models by RT-PCR and the concentration of TNF-α in cell model’s supernanant by ELISA. At the same time, the effects of sense oligodeoxynucleotide (SODN) and randomized nonsense-sequence oligodeoxynucleotide (RODN) were detected and compared with AODN. Results:The specific AODN decreased the production of CVB3 and CVB5 antigens in persistent infection cell models and this effect depended on the AODN’ s dose. After reacted by AODN, the degree of CPE on Vero cells caused by supernanat of cell models was inhibited. The viral titers in culture supertanant decreased and viral RNA couldn’t be detected by RT-PCR. The concentration of TNF-α of cell models increased. ADON which was synthesized according to genes of CVB3 also had strongly inhibited to CVB5 persistent infection. The inhibition of AODN was stronger than SODN and RODN. Conclution: AODN can inhibit persistent infections of CVB3 and CVB5 in vitro and induce cells to produce TNF-α. This effect has sense specific but there is cross inhibition between the same viral groups. Maybe it is related to the homogenous genetic sense of CVB3 and CVB5.