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1 2 3 4 5 6Lanes:1 – negative control of RT-PCR2-4 – viral RNA after incubation of influenza virus A with 25 µMsolution of K-L 1 -K (lane 2), K-L 2 -K (lane 3) and E-L 2 -E (lane 4) for1 hour at 37 0 C5 – influenza A virus RNA6 – positive control of RT-PCRFig. 2. Results of electrophoresis in 1.5% agarose gel of RT-PCR products.(%)viral reproduction (%)100806040200100806040200influnza virus A H1N1 (Aichi/1/68)Virus control S-L2-S K-L1-K K-L2-K E-L2-E H-L1-Hinfluenza virus B/Minsk/140/07.Fig. 3. Results of ELISA of the influenza virusA and B antigens after treatment of infectedMDCK cells with peptidomimetics. Viralreproduction (%) = (OD in the presence ofthe artificial RNases/ OD of viralcontrol)x100%. (Top) ELISA of influenzavirus A H1N1 antigen after treatment with 25µg/ml artificial RNases; (Middle) ELISA ofinfluenza virus B antigen after treatment with3.125 µg/ml artificial RNases; (Bottom)ELISA of influenza virus A H3N2 antigenafter treatment with 3.125 µg/ml artificialRNases. Simultaneous infection and additionof artificial RNases (gray columns) ortreatment of MDCK cells with artificialRNases in 60 min before infection (blackcolumn).Virus control K-L2-K E-L1-E E-L2-E S-L1-S S-L2-Sinfluenza virus A H3N2 (Ivachevichi/115/07)Inhibition of influenza virus A (H1N1 (strain100Aichi 1/68) and H3N2 (strain Ivachevichi/80115/07)) and B (strain Minsk/140/07) wasestimated in hemagglutination (HA), enzymelinkedimmunosorbent assay (ELISA) and6040reverse transcription (RT) with subsequentPCR. Treatment of extracellular virions of20both influenza virus A and B with peptidomimetics(3.125-25 µg/ml) for periods from0Virus K-L1-K K-L2-K E-L1-E E-L2-E S-L1-S S-L2-Scontrol30 min to 18 hours resulted in negative HAand ELISA (Figure 3) in 1-3 days postinfection. RT-PCR revealed complete cleavage of the influenza virus RNA before infection(Figure 2).One should note that HA and ELISA titers corresponding to the virus antigen amountsremained the same before infection. Additions of peptidomimetics in culture media ofinfected MDCK cells in a few hours post infection did not cause essential viral RNAdegradation, inhibition of HA and ELISA, as well as infectivity reduction. Taken together,the data proved RNase activity of new artificial RNases, absence of influenza virus surfaceantigens (HA and neuraminidase) modifications and penetration of peptidomimetics intoextracellular virions but not within infected cells.AcknowledgmentsThis work was supported by RFBR-09-04-01483, Integration grant of SB RAS-83, Integration grant ofSB RAS-88.References1. Koroleva, L., et al. Russ. Chem. Bull. 54, 2682-2691 (2005).2. Konevetz, D., et al. Russ. J. Bioorg. Chem. 28, 331-341 (2002).3. Kuznetsova, I., et al. Russ. Chem. Bull. 53, 455-462 (2004).viral reproduction (%)337