Additive Effects of Water-Soluble Propolis (Greit 120) and Human Interferon Alfa (HuIFN-αN3) against Influenza Viruses in Vitro -David Publishing Company
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1. Croatian Institute of Experimental and Translation Oncology, Koledinećka 3, Zagreb 10040, Croatia
2. Institute of Biomedical Sciences, Medical Faculty of Maribor, Taborska 8, Maribor 2000, Slovenia
3. Medex D.o.o., Linhartova 49b, Ljubljana 1000, Slovenia
4. Chengdujska 4, Ljubljana 1000, Slovenia
5. Institute of Microbiology and Immunology, Medical Faculty, Zaloška 4, Ljubljana 1105, Slovenia
6. Division of Molecular Biology, Ruđer Bošković Institute, Bijenička 54, Zagreb 10000, Croatia
7. Trg Sv. Ivana 5, Kloštar Ivanić 10312, Croatia
8. Department of Life Sciences, University of Modena & Reggio Emilia, Modena 41125, Italy
9. BNatural, Via Gran Sasso 33, Corbetta-MI 20011, Italy


Influenza virus affects the respiratory tract in humans causing a range of distinct manifestations including fever, nasal secretions, cough, headaches, muscle pain and pneumonia, which could become violent and severe. It was found that influenza A viruses remain resistant to amantadine and rimantadin with high level of oseltamvir resistance. Therefore, there is a need for constant improvement of drugs active against resistant influenza viruses. Propolis has anti-influenza activity both in vitro and in vivo. Human leukocyte interferon (HuIFN-αN3) is a multi-subtype protein that displays an antiviral activity against influenza virus. In this study we elucidated the anti-influenza activity of the mixes of water-soluble propolis (WSP) (Greit 120) and HuIFN-αN3 at different ratios. Greit 120 polyphenols, total phenol acids and bioflavonoid were characterized by HPLC-UV-ESI-MS504971 and HuIFN-αN3 by reverse-phase high-performance liquid chromatography (RP-HPLC). Influenza A and B viruses were separately added to the LLC-MK2 cells treated with WSP (Greit 120) and HuIFN-αN3 alone or in proportions 1:1, 1:2 and 2:1. Plates were incubated and cytopathic effect was determined. The best results (ID50) were obtained with the mix of 10% WSP and HuIFN-αN3 in proportion 1:2, showing ID50 at 12 ± 0.2 µg/mL and 19 ± 0.6 µg/mL for influenza A and B viruses, respectively. When comparing anti-influenza activity of WSP (Greit 120)/HuIFN-αN3 with that of ribavirin, it was found that 1:2 was the optimal ratio for WSP (Greit 120)/HuIFN-αN3 (0.5 and 0.6 for influenza A and B, respectively). This new formulation of WSP (Greit 120) and HuIFN-αN3, showing better anti-Influenza activity, will definitely improve its application in flu infections.


Water-soluble propolis, Greit 120, HuIFN-αN3, anti-influenza activity, ribavirin, ID50.

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