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Medycyna Doświadczalna i Mikrobiologia 2016, 1(68): 64-71

Wpływ interferonu-α i pranobeksu inozyny na hamowanie replikacji ludzkich wirusów RNA in vitro
[Interferon-α and inosine pranobex-mediated inhibition of replication of human RNA viruses in vitro]

Anna Majewska, Witold Lasek, Maciej Przybylski, Tomasz Dzieciątkowski, Grażyna Młynarczyk

STRESZCZENIE

Celem badania była ocena wpływu interferonu-α (IFN-α) oraz pranobeksu inozyny (PI), na miana zakaźne wirusów RNA: wirusa Coxackie A16, enterowirusa 71, oraz wirusa paragrypy 4 w warunkach in vitro. Oceny miana zakaźnego wirusów przeprowadzono według metody Reeda-Muencha. Miano zakaźne wyrażono w log10 TCID50/ml. Wirus Coxsackie A16 okazał się najwrażliwszy na działanie zastosowanych inhibitorów. Aplikacja 1000 IU/ml IFN-α łącznie z PI do hodowli komórek linii A549 zakażonych tym wirusem zredukowała IC50 o 41,7 %.

ABSTRACT

Introduction: Interferon- α (IFN-α), produced by immune cells, exhibits pleiotropic antiviral activity. Inosine pranobex (PI), a synthetic derivative of a purine, shows direct antiviral activity, and also acts indirectly, by activation of immune cells. The aim of this study was to evaluate an in vitro inhibition of Coxackievirus A16 (CA16), enterovirus 71 (EV71) and human parainfluenza virus 4 (HPIV-4) replication by PI in combination with IFN-α.

Materials and Methods: In the present study we evaluated an in vitro effect of interferon-α and inosine pranobex on replication of RNA viruses: CA-16, EV71, HPIV-4. Antiviral effects of IFN-α and IP were assessed by phenotypic assays. The yield reduction assay (YRA), which evaluates the ability of the compounds to inhibit virus multiplication in cell cultures, was applied. The Reed–Muench statistical method was used to determine the 50% end point (IC50) .

64 Nr 1 A. Majewska i inni

Results: Our studies have shown that combination of IFN-α and inosine pranobex display higher efficacy than treatment with either compound alone, and suggest synergy that may increase therapeutic effectiveness. The reduction of the average viral titers of EV71, CA-16 and HPIV-4 in A549 cell culture after applying 400 µg/mL IP and IFN-α (1000 IU/mL), in comparison to the viral titer in the control was reduced by 1,76 log10 TCID50/ml, o 3,00 log10 TCID50/ml , and 1,60 log10 TCID50/ml respectively. The antiviral activity of the tested compounds was also analyzed on the basis of IC50 values. Application of 1000 IU/ml IFN-α, with PI after infection of A549 cells with mention above viruses reduced the IC50 by 3,5%, 41,3% and 29% respectively.

Conclusions: Our study demonstrated that enhanced antiviral activity was observed when cells infected with RNA viruses were treated with combination of IFN-α and IP. The effectiveness of IFN-α and IP under these conditions has not been previously reported. CA16 virus turned out to be the most sensitive to the action of used inhibitors.



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