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. 2019 Jul 22;1(6):e000041.
doi: 10.1099/acmi.0.000041. eCollection 2019.

Brefeldin A and Cytochalasin B reduce dengue virus replication in cell cultures but do not protect mice against viral challenge

Kleber Juvenal Silva Farias  1   2 , Paula Renata Lima Machado  3 , Renato Ferreira de Almeida Júnior  3 , Benedito Antônio Lopes da Fonseca  1
Affiliations

Brefeldin A and Cytochalasin B reduce dengue virus replication in cell cultures but do not protect mice against viral challenge

Kleber Juvenal Silva Farias et al. Access Microbiol. .

Abstract

Background: Dengue is an emerging arboviral disease caused by dengue virus (DENV). DENV belongs to the family Flaviviridae and genus Flavivirus. No specific anti-DENV drugs are currently available.

Methods: We investigated the antiviral activity of Brefeldin A (BFA) and Cytochalasin B (CB) against this infection. The drugs BFA and CB were used in the in vitro treatment of dengue-2 virus (DENV-2) infections in Vero cell cultures and in protection from lethality by post-challenge administration in Swiss mice. Viral load was quantified by qRT-PCR and plaque assay in Vero cell cultures, post-infection, treated or not with the drugs. Post-challenge drug levels were evaluated by survival analysis.

Results: Our results indicate that doses of 5 µg ml-1 of BFA and 10 µg ml-1 of CB are not toxic to the cells and induce a statistically significant inhibition of DENV-2 replication in Vero cells when compared to control. No BFA- or CB-treated mice survived the challenge with DENV-2.

Conclusion: These data suggest that BFA and CB have an antiviral action against DENV-2 replication in Vero cell culture, but do not alter infected mice mortality.

Keywords: BFA; CB; Cell Culture; DENV-2; Mice; Viral load monitoring; qRT-PCR.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
The two-dimensional chemical structure of brefeldin A (a) and cytochalasin B (b). Source: PubChem. URL: https://pubchem.ncbi.nlm.nih.gov.
Fig. 2.
Fig. 2.
Cytotoxicity of the drugs (BFA and CB) and organic solvents (DMSO and ethanol) on Vero cells. BFA, brefeldin A; CB, cytochalasin B; DMSO, dimethyl sulfoxide.
Fig. 3.
Fig. 3.
Action of BFA or CB, added 1 h post-infection, on DENV-2 replication in Vero cells. (a,b) The viral RNA present in the culture supernatants of Vero cells infected with DENV-2, treated or untreated with BFA or CB 1 h post-infection, was extracted and analysed by qRT-PCR. (c,d) Culture supernatants of Vero cells infected with DENV-2, treated or untreated with BFA or CB 1 h post-infection, were analysed by plaque assay. The results represent the average values of the viral RNA copy number (P<0.001). BFA, brefeldin A; CB, cytochalasin B; DENV-2, dengue-2 virus; qRT-PCR, real time quantitative reverse transcriptase PCR.
Fig. 4.
Fig. 4.
Action of BFA or CB, added at 24 h intervals, on DENV-2 replication in Vero cells. (a,b) The viral RNA present in the culture supernatants of Vero cells infected with DENV-2, treated or untreated with BFA or CB 1 h post-infection, and at 24 h intervals, was extracted and analysed by qRT-PCR. (c,d) Culture supernatants of Vero cells infected with DENV-2, treated or untreated with BFA or CB 1 h post-infection, and at 24 h intervals, were analysed by plaque assay. The results represent the average values of the viral RNA copy number (P<0.001). BFA, brefeldin A; CB, cytochalasin B; DENV-2, dengue-2 virus; qRT-PCR, real time quantitative reverse transcriptase PCR.
Fig. 5.
Fig. 5.
Survival curves of mice infected with DENV-2. (a,b) Swiss mice received BFA at 26.3 mg kg−1 or CB at 1.2 mg kg−1 2 h post-infection. Survival analysis was performed by observing these animals for 21 days. Note that the mice of the positive control group died by the 10th day and those receiving BFA or CB and virus died on the 11th day. BFA, brefeldin A; CB, cytochalasin B; DENV-2, dengue-2 virus
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