Mikrobiol. Z. 2020; 82(4):71-79.
doi: https://doi.org/10.15407/microbiolj82.04.071

Antiviral Activity of Extracts from Wild Grasses against Epstein-Barr Virus and
Induction of Apoptosis in Epstein-Barr Virus-Positive Lymphoblastoid Cells

A.V. Holovan, K.S. Naumenko, F.V. Muchnyk, G.V. Baranova,
L.B. Zelena, S.D. Zagorodnya

Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

Epstein-Barr virus (EBV) belongs to the human herpesvirus family that infects more than 90 % of the population. EBV is associated with a number of lymphoproliferative and autoimmune diseases. Usage of the drugs, which would not only inhibit the reproduction of the virus, but also would stimulate the elimination of tumor cells, is important for the treatment of virus-associated tumors. The purpose. In the current research the antiviral effects and apoptotic activity of the herbal drugs proteflazid and neoflazid were studied on the models of latent, acute and chronic EBV infection in Raji and B95-8 lymphoblastoid cells. Methods. The investigations were performed in Raji (virus non-productive) and B95-8 (virusproductive) lymphoblastoid cells. Proteflazid and neoflazid, plant drugs, were studied, acycloguanosine was used as a reference drug. Trypan blue staining (TBS) method and MTT-assay were used to study of cell viability. Antiviral activity was estimated by real time polymerase chain reaction (RT-PCR). Apoptotic cells were detected using flow cytometry method. Results. The investigations showed that neoflazid was more toxic towards Raji cells than proteflazid: the cytotoxic concentration (CC50) indexes were 8 μg/ml and 36 μg/ml respectively. Toxicities of these compounds in B95-8 cells were almost the same and their CC50 indexes were close to 25 μg/ml. Both drugs showed high antiviral activity against EBV lytic infection in Raji cells and effective concentration (EC50) was 0.02 and 0.083 μg/ml for proteflazid and neoflazid, respectively, and selectivity indexes (SI) were 1800 and 96. These compounds were less effective in B95-8 cells and even at the concentration of 10 μg/ml inhibited virus replication only by 10–19%. We checked an ability of proteflazid to induce apoptosis and found that the drug at cytotoxic concentration (30 μg/ml) stimulated the apoptotic death of 70% cells in latent and lytic EBV infection. The non-toxic concentration (5 μg/ml) induced apoptosis 30% of cells. Conclusions. Thus, our research showed that both plant preparations proteflazid and neoflazid possessed high antiviral activity under acute EBV infection. Proteflazid, in addition, induced dose-depending apoptosis in EBV-positive lymphoblastoid cells.

Keywords: Epstein-Barr virus, proteflazid, neoflazid, antiviral activity, apoptosis.

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