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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 EV71 production.(a) RD cells were pre-incubated with Amphotericin B at increasing concentrations for 2 h, and then infected with EV71 at an MOI of 0.05. After 24 h, culture medium was collected and viral titers were determined in plaque assays. The data represent 3 independent experiments, and error bars represent SD (*P < 0.05, **P < 0.01, t test). (b) Cytotoxicity of Amphotericin B on RD cells was measured by performing CellTiter-Glo® Luminescent Cell Viability Assay. (c) Effect of Amphotericin B on the production of EV71 in 293 cells. The data represent 3 independent experiments, and error bars represent SD (**P < 0.01, t test). (d) Effects of Amphotericin B on the viability of 293 cells. (e) EC50 and IC50 of Amphotericin B on EV71 in RD and 293 cells.
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f2: Amphotericin B inhibits EV71 production.(a) RD cells were pre-incubated with Amphotericin B at increasing concentrations for 2 h, and then infected with EV71 at an MOI of 0.05. After 24 h, culture medium was collected and viral titers were determined in plaque assays. The data represent 3 independent experiments, and error bars represent SD (*P < 0.05, **P < 0.01, t test). (b) Cytotoxicity of Amphotericin B on RD cells was measured by performing CellTiter-Glo® Luminescent Cell Viability Assay. (c) Effect of Amphotericin B on the production of EV71 in 293 cells. The data represent 3 independent experiments, and error bars represent SD (**P < 0.01, t test). (d) Effects of Amphotericin B on the viability of 293 cells. (e) EC50 and IC50 of Amphotericin B on EV71 in RD and 293 cells.

Mentions: We next measured the levels of EV71 production under Amphotericin B treatment. Titers of EV71 that was produced by RD cells or 293 cells were determined in the plaque assay. Figure 2a show that Amphotericin B drastically reduced the production of EV71 in RD cells with an EC50 (50% effective concentration) of 1.75 ± 0.05 μM. Similar trend of inhibition by Amphotericin B was observed in 293 cells with an EC50 of 0.32 ± 0.02 μM (Fig. 2c). To exclude the possibility that the inhibition of viral production is a result of Amphotericin B-mediated cytotoxicity, we measured cell viability under Amphotericin B treatment. The results showed that Amphotericin B had a CC50 (50% cytotoxic concentration) of 7.37 ± 0.07 μM in RD cells and 14.5 ± 0.05 μM in 293 cells, which are much higher than the EC50 values (Fig. 2b,d). These results demonstrate that Amphotericin B potently inhibits EV71 infection in both RD and 293 cells.


Amphotericin B Inhibits Enterovirus 71 Replication by Impeding Viral Entry
Amphotericin B inhibits EV71 production.(a) RD cells were pre-incubated with Amphotericin B at increasing concentrations for 2 h, and then infected with EV71 at an MOI of 0.05. After 24 h, culture medium was collected and viral titers were determined in plaque assays. The data represent 3 independent experiments, and error bars represent SD (*P < 0.05, **P < 0.01, t test). (b) Cytotoxicity of Amphotericin B on RD cells was measured by performing CellTiter-Glo® Luminescent Cell Viability Assay. (c) Effect of Amphotericin B on the production of EV71 in 293 cells. The data represent 3 independent experiments, and error bars represent SD (**P < 0.01, t test). (d) Effects of Amphotericin B on the viability of 293 cells. (e) EC50 and IC50 of Amphotericin B on EV71 in RD and 293 cells.
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f2: Amphotericin B inhibits EV71 production.(a) RD cells were pre-incubated with Amphotericin B at increasing concentrations for 2 h, and then infected with EV71 at an MOI of 0.05. After 24 h, culture medium was collected and viral titers were determined in plaque assays. The data represent 3 independent experiments, and error bars represent SD (*P < 0.05, **P < 0.01, t test). (b) Cytotoxicity of Amphotericin B on RD cells was measured by performing CellTiter-Glo® Luminescent Cell Viability Assay. (c) Effect of Amphotericin B on the production of EV71 in 293 cells. The data represent 3 independent experiments, and error bars represent SD (**P < 0.01, t test). (d) Effects of Amphotericin B on the viability of 293 cells. (e) EC50 and IC50 of Amphotericin B on EV71 in RD and 293 cells.
Mentions: We next measured the levels of EV71 production under Amphotericin B treatment. Titers of EV71 that was produced by RD cells or 293 cells were determined in the plaque assay. Figure 2a show that Amphotericin B drastically reduced the production of EV71 in RD cells with an EC50 (50% effective concentration) of 1.75 ± 0.05 μM. Similar trend of inhibition by Amphotericin B was observed in 293 cells with an EC50 of 0.32 ± 0.02 μM (Fig. 2c). To exclude the possibility that the inhibition of viral production is a result of Amphotericin B-mediated cytotoxicity, we measured cell viability under Amphotericin B treatment. The results showed that Amphotericin B had a CC50 (50% cytotoxic concentration) of 7.37 ± 0.07 μM in RD cells and 14.5 ± 0.05 μM in 293 cells, which are much higher than the EC50 values (Fig. 2b,d). These results demonstrate that Amphotericin B potently inhibits EV71 infection in both RD and 293 cells.

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