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. 2013 Feb:52:207-15.
doi: 10.1016/j.fct.2012.11.006. Epub 2012 Nov 23.

Inhibition of herpes simplex virus type 1 with the modified green tea polyphenol palmitoyl-epigallocatechin gallate

Aline de Oliveira  1 , Sandra D Adams, Lee H Lee, Sean R Murray, Stephen D Hsu, Jeffrey R Hammond, Douglas Dickinson, Ping Chen, Tin-Chun Chu
Affiliations

Inhibition of herpes simplex virus type 1 with the modified green tea polyphenol palmitoyl-epigallocatechin gallate

Aline de Oliveira et al. Food Chem Toxicol. 2013 Feb.

Abstract

Green tea polyphenol epigallocatechin gallate (EGCG) is a strong antioxidant that has previously been shown to reduce the number of plaques in HIV-infected cultured cells. Modified EGCG, palmitoyl-EGCG (p-EGCG), is of interest as a topical antiviral agent for herpes simplex virus (HSV-1) infections. This study evaluated the effect of p-EGCG on HSV-infected Vero cells. Results of cell viability and cell proliferation assays indicate that p-EGCG is not toxic to cultured Vero cells and show that modification of the green tea polyphenol epigallocatechin gallate (EGCG) with palmitate increases the effectiveness of EGCG as an antiviral agent. Furthermore, p-EGCG is a more potent inhibitor of herpes simplex virus 1 (HSV-1) than EGCG and can be topically applied to skin, one of the primary tissues infected by HSV. Viral binding assay, plaque forming assay, PCR, real-time PCR, and fluorescence microscopy were used to demonstrate that p-EGCG concentrations of 50 μM and higher block the production of infectious HSV-1 particles. p-EGCG was found to inhibit HSV-1 adsorption to Vero cells. Thus, p-EGCG may provide a novel treatment for HSV-1 infections.

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Figures

Figure 1
Figure 1
Cell viability of Vero cells treated with different concentrations of p-EGCG. The numbers represent the mean of three replicates and the y-error bars represent SD.
Figure 2
Figure 2
Cell proliferation studies of Vero cells treated with different concentrations (0, 50 and 75 µM) of EGCG and p-EGCG using MTS assay. The means of results of triplicate tests of EGCG treated samples (blue) and p-EGCG treated samples (red) represent the average of triplicate trials. The y-error bars represent SD.
Figure 3
Figure 3
PFU of HSV-1/Vero, HSV-1 treated 50 µM EGCG/Vero, HSV-1 treated 50 µM p-EGCG/Vero. The numbers represent the mean of triplicates and the y-error bars represent SD.
Figure 4
Figure 4
PFU of HSV-1 treated with different concentrations of p-EGCG. The numbers represent the mean of triplicate trials and the y-error bars represent SD.
Figure 5
Figure 5
Fluorescence microscopic observations of control Vero cells and p-EGCG treated HSV-1 infection of Vero Cells. 5.1. Control Vero cells monolayer (400×). (A) Phase contrast; (B) DAPI stain; (C) GFP. 5.2. GFP expression and 5.3. DAPI stain at 8–12 hours post-infection for HSV-1 (+/− p-EGCG) of Vero cells at (A) 8 hours no p-EGCG (A’) 8 hours + p-EGCG; (B) 10 hours no p-EGCG; (B’) 10 hours + p-EGCG; (C) 12 hours no p-EGCG; (C’) 12 hours + p-EGCG. The arrows show the granulation and loss of margins in the HSV-1 infected cells.
Figure 5
Figure 5
Fluorescence microscopic observations of control Vero cells and p-EGCG treated HSV-1 infection of Vero Cells. 5.1. Control Vero cells monolayer (400×). (A) Phase contrast; (B) DAPI stain; (C) GFP. 5.2. GFP expression and 5.3. DAPI stain at 8–12 hours post-infection for HSV-1 (+/− p-EGCG) of Vero cells at (A) 8 hours no p-EGCG (A’) 8 hours + p-EGCG; (B) 10 hours no p-EGCG; (B’) 10 hours + p-EGCG; (C) 12 hours no p-EGCG; (C’) 12 hours + p-EGCG. The arrows show the granulation and loss of margins in the HSV-1 infected cells.
Figure 5
Figure 5
Fluorescence microscopic observations of control Vero cells and p-EGCG treated HSV-1 infection of Vero Cells. 5.1. Control Vero cells monolayer (400×). (A) Phase contrast; (B) DAPI stain; (C) GFP. 5.2. GFP expression and 5.3. DAPI stain at 8–12 hours post-infection for HSV-1 (+/− p-EGCG) of Vero cells at (A) 8 hours no p-EGCG (A’) 8 hours + p-EGCG; (B) 10 hours no p-EGCG; (B’) 10 hours + p-EGCG; (C) 12 hours no p-EGCG; (C’) 12 hours + p-EGCG. The arrows show the granulation and loss of margins in the HSV-1 infected cells.
Figure 6
Figure 6
Binding assay of HSV-1, with or without 75 µM p-EGCG, with Vero cells. The numbers represent the mean of three trials and the y-error bars represent SD.
Figure 7
Figure 7
Gel electrophoresis of PCR products. Top: PCR of HSV-1 glycoprotein D gene US6. 100 ng of DNA extracted from HSV-1/Vero cells was added to each PCR reaction. Middle: PCR of HSV-1 GFP gene UL46. Bottom: PCR of HSV-1 VP11/12 tegument gene UL46. Lane 1: HSV-1/Vero; Lane 2: 75 µM EGCG-HSV-1/Vero; Lane 3: 75 µM palmitoyl-EGCG-HSV-1/Vero; Lane 4: Vero cells only; Lane 5: Negative control.
Figure 8
Figure 8
Percentage relative to control of concentration of US6 gene in EGCG-HSV-1/Vero and p-EGCG-HSV-1/Vero cells relative to HSV-1/ Vero cells with no EGCG or p-EGCG based on RT-PCR. Percentage calculated based on the relative expression with comparative Ct (n=3). Y-error bars indicate SD.
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References

    1. Akhtar J, Shukla D. Viral entry mechanisms: cellular and viral mediators of herpes simplex virus entry. FEBS J. 2009;276(24):7228–7236. - PMC - PubMed
    1. Akkarawongsa R, Pocaro NE, et al. Multiple peptides homologous to herpes simplex virus type 1 glycoprotein B inhibit viral infection. Antimicrob Agents Chemother. 2009;53(3):987–996. - PMC - PubMed
    1. Babich H, Krupka ME, et al. Differential in vitro cytotoxicity of (−)-epicatechin gallate (ECG) to cancer and normal cells from the human oral cavity. Toxicol In Vitro. 2005;19(2):231–242. - PubMed
    1. Belshe RB, Leone PA, et al. Efficacy results of a trial of a herpes simplex vaccine. N Engl J Med. 2012;366(1):34–43. - PMC - PubMed
    1. Bernstein DI, Stanberry LR. Herpes simplex virus vaccines. Vaccine. 1999;17(13–14):1681–1689. - PubMed

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