Determination The Antiviral Activity Of Novaferon In Mouse L929Cells

Objective:To detect whether Novaferon possess the species specificity; to determine the antiviral activity of Novaferon in mouse cells L929which were infected by VSV; to discuss the possibility of Novaferon on preclinical research in the mouse models of human diseases in the future.Methods:Propagate IFN-sensitive L929fibroblasts and prepare the cell suspension. Seed the cells in a96-well flat-bottom microtiter plate, and incubate the plate in a37℃、5%CO2humidified incubator overnight. With different concentration for the start, use the continuous on-fold dilution of the Novaferon or MuIFN-α1and add the cells to each well of the96-well plate. Set the normal cells group and culture for24hours. Dilute VSV in supplemented DMEM to an appropriate concentration. After abandon the culture media, add virus to each well of the Novaferon group and the MuIFN-a1group, respectively (MOI:0.1) Set the normal cells group and the virus control group, culture for36hours. Remove the supernatants, use0.85%NaCl to rinse thoroughly and pat dry. Add each well of Cell staining fluid. Incubate at room temperature for2hours. Rinse plate with tap water and allow to dry. Add each well with2-methoxyethanol for45min in order to destain. Read the samples spectrophotometrically at550nm. Repeat the study for three times. Average the results and calculate the protection ratio. With the logarithm of drug concentration as horizontal axis and the protection ratio as vertical axis to draw the regression analysis diagram of Novaferon and MuIFN-α1in L929-VSV system, respectively. Calculate the EC50of these two kinds of drug by linear regression.Results:①The values of OD550of the Novaferon and MuIFN-α1were improved significantly. The protection ratio of Novaferon and MuIFN-a1increased with the up-regulation of the concentration of the drug. And there was a positive correlation between them (Novaferon:r=0.972, p<0.001; MuIFN-α1:r=0.950, p<0.001). At the same concentration, the protection ratio of MuIFN-α1was higher than Novaferon in mouse L929cells when we ovserved the regression analysis diagram. This implied that the antiviral activity of the MuIFN-a1was higher than Novaferon.②The value of EC50of Novaferon was170.752pg, and the MuIFN-a1was38.810pg. Then we can calculate the antiviral activity of Novaferon in L929-VSV system was6.32×106IU/mg. It confirmed that Novaferon own a good antiviral activity in mouse L929cells which were infected with VSV. And it possesses an obviously interspecific cross effect. Conclusion:①Novaferon and MuIFN-α1are both possess the protective effect in mouse L929cells which were infected by VSV, and there was a positive correlation between them;②At the same concentration, the protection ratio of MuIFN-a1was higher than Novaferon in mouse L929cells, and the antiviral activity of MuIFN-α1was higher than Novaferon;③Novaferon possesses an obviously interspecific cross effect in VSV virus infected mouse L929cells. It can be used in the research of Novaferon animal experiments in mouse.