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Amphotericin B Inhibits Enterovirus 71 Replication by Impeding Viral Entry

View Article: PubMed Central - PubMed

ABSTRACT

Enterovirus 71 (EV71) infection causes hand-foot-and-mouth disease that leads to cardiopulmonary complications and death in young children. There is thus an urgent need to find new treatments to control EV71 infection. In this study, we report potent inhibition of EV71 by a polyene antibiotic Amphotericin B. Amphotericin B profoundly diminished the expression of EV71 RNA and viral proteins in the RD cells and the HEK293 cells. As a result, EV71 production was inhibited by Amphotericin B with an EC50 (50% effective concentration) of 1.75 μM in RD cells and 0.32 μM in 293 cells. In addition to EV71, EV68 was also strongly inhibited by Amphotericin B. Results of mechanistic studies revealed that Amphotericin B targeted the early stage of EV71 infection through impairing the attachment and internalization of EV71 by host cells. As an effective anti-fungi drug, Amphotericin B thus holds the promise of formulating a novel therapeutic to treat EV71 infection.

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Amphotericin B inhibits the early stage of EV71 infection.(a) RD cells were infected with EV71 at an MOI of 4 in the absence or presence of 2 μM Amphotericin B. Levels of EV71 proteins in the infected cells were determined by Western blotting at 2, 4, 6, 8, 10, and 12 h post infection. (b) 293 cells were infected with EV71 at an MOI of 25 in the absence or presence of 1 μM Amphotericin B. Full-length blots are presented in Supplementary Fig. 3. (c) EV71 RNA in RD cells was quantified by qRT-PCR at 2, 4, 6, 8, 10 hpi. The data represented the copy number of EV71 RNA. Results shown are the average of three independent experiments. Error bars represent SD (*P < 0.05, **P < 0.01, t test).
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f4: Amphotericin B inhibits the early stage of EV71 infection.(a) RD cells were infected with EV71 at an MOI of 4 in the absence or presence of 2 μM Amphotericin B. Levels of EV71 proteins in the infected cells were determined by Western blotting at 2, 4, 6, 8, 10, and 12 h post infection. (b) 293 cells were infected with EV71 at an MOI of 25 in the absence or presence of 1 μM Amphotericin B. Full-length blots are presented in Supplementary Fig. 3. (c) EV71 RNA in RD cells was quantified by qRT-PCR at 2, 4, 6, 8, 10 hpi. The data represented the copy number of EV71 RNA. Results shown are the average of three independent experiments. Error bars represent SD (*P < 0.05, **P < 0.01, t test).

Mentions: Amphotericin B has been shown to inhibit different viruses at various stages of their life cycles3337. We therefore examined which step of EV71 lifecycle was affected by Amphotericin B. RD cells or 293 cells were infected with EV71 in the absence or presence of Amphotericin B, and viral protein expression was measured by Western blotting. Expression of EV71 VP1 and VP2 was decreased at the earliest detectable time point (6 hpi in RD and 8 hpi in 293) (Fig. 4a,b). We next assessed the inhibition by quantifying the viral RNA through quantitative RT-PCR at 2, 4, 6, 8, and 10 h post infection (Fig. 4c). The results showed that the amount of EV71 RNA in the infected cells was reduced at 2 hpi under Amphotericin B treatment. These data suggest that Amphotericin B impedes an early step of EV71 life cycle.


Amphotericin B Inhibits Enterovirus 71 Replication by Impeding Viral Entry
Amphotericin B inhibits the early stage of EV71 infection.(a) RD cells were infected with EV71 at an MOI of 4 in the absence or presence of 2 μM Amphotericin B. Levels of EV71 proteins in the infected cells were determined by Western blotting at 2, 4, 6, 8, 10, and 12 h post infection. (b) 293 cells were infected with EV71 at an MOI of 25 in the absence or presence of 1 μM Amphotericin B. Full-length blots are presented in Supplementary Fig. 3. (c) EV71 RNA in RD cells was quantified by qRT-PCR at 2, 4, 6, 8, 10 hpi. The data represented the copy number of EV71 RNA. Results shown are the average of three independent experiments. Error bars represent SD (*P < 0.05, **P < 0.01, t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5016833&req=5

f4: Amphotericin B inhibits the early stage of EV71 infection.(a) RD cells were infected with EV71 at an MOI of 4 in the absence or presence of 2 μM Amphotericin B. Levels of EV71 proteins in the infected cells were determined by Western blotting at 2, 4, 6, 8, 10, and 12 h post infection. (b) 293 cells were infected with EV71 at an MOI of 25 in the absence or presence of 1 μM Amphotericin B. Full-length blots are presented in Supplementary Fig. 3. (c) EV71 RNA in RD cells was quantified by qRT-PCR at 2, 4, 6, 8, 10 hpi. The data represented the copy number of EV71 RNA. Results shown are the average of three independent experiments. Error bars represent SD (*P < 0.05, **P < 0.01, t test).
Mentions: Amphotericin B has been shown to inhibit different viruses at various stages of their life cycles3337. We therefore examined which step of EV71 lifecycle was affected by Amphotericin B. RD cells or 293 cells were infected with EV71 in the absence or presence of Amphotericin B, and viral protein expression was measured by Western blotting. Expression of EV71 VP1 and VP2 was decreased at the earliest detectable time point (6 hpi in RD and 8 hpi in 293) (Fig. 4a,b). We next assessed the inhibition by quantifying the viral RNA through quantitative RT-PCR at 2, 4, 6, 8, and 10 h post infection (Fig. 4c). The results showed that the amount of EV71 RNA in the infected cells was reduced at 2 hpi under Amphotericin B treatment. These data suggest that Amphotericin B impedes an early step of EV71 life cycle.

View Article: PubMed Central - PubMed

ABSTRACT

Enterovirus 71 (EV71) infection causes hand-foot-and-mouth disease that leads to cardiopulmonary complications and death in young children. There is thus an urgent need to find new treatments to control EV71 infection. In this study, we report potent inhibition of EV71 by a polyene antibiotic Amphotericin B. Amphotericin B profoundly diminished the expression of EV71 RNA and viral proteins in the RD cells and the HEK293 cells. As a result, EV71 production was inhibited by Amphotericin B with an EC50 (50% effective concentration) of 1.75&thinsp;&mu;M in RD cells and 0.32&thinsp;&mu;M in 293 cells. In addition to EV71, EV68 was also strongly inhibited by Amphotericin B. Results of mechanistic studies revealed that Amphotericin B targeted the early stage of EV71 infection through impairing the attachment and internalization of EV71 by host cells. As an effective anti-fungi drug, Amphotericin B thus holds the promise of formulating a novel therapeutic to treat EV71 infection.

No MeSH data available.


Related in: MedlinePlus