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Lethal mutagenesis of the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV)
Carmen M Ruiz-Jarabo1, Calvin Ly, Esteban Domingo
1Centro de Biologia Molecular Severo Ochoa (CSIC, UAM), Universidad Autonoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
Abstract:
Passage of the prototypic arenavirus lymphocytic choriomenigitis virus (LCMV) in cultured cells in the presence of the mutagenic agent 5-fluorouracil (FU) resulted in efficient and systematic virus extinction under conditions that did not significantly affect cell survival. FU-mediated extinction of LCMV was associated with 3.6- to 10-fold increases in the mutation frequencies for the three viral genes examined, but with only very modest effects on virus replication and transcription during a single round of infection. Likewise, FU did not affect expression of a LCMV minigenome. In contrast, the well documented antiviral effect of ribavirin against LCMV was not associated with significant increases in virus mutation frequencies, but rather with a dramatic inhibition of both viral RNA synthesis and LCMV minigenome expression. Mutagen induced viral extinction has been recently reported for positive strand RNA viruses polio and foot-and-mouth disease, and the lentivirus HIV-1. Our findings indicate that lethal mutagenesis can be effective also against LCMV, a negative strand RNA virus. Moreover, FU treatment prevented the establishment of LCMV persistent infection in mice deficient in B and T cells, suggesting the feasibility in vivo of lethal mutagenesis as a novel antiviral strategy.
Insights
The antiviral 5-fluorouracil (FU) effectively eliminated lymphocytic choriomenigitis virus (LCMV) by inducing lethal mutagenesis. This novel strategy shows promise for treating LCMV infections in vivo.
Area of Science:
- Virology
- Molecular Biology
- Antiviral Research
Background:
- Lymphocytic choriomenigitis virus (LCMV) is a prototypic arenavirus.
- Existing antiviral strategies may have limitations.
- The potential of lethal mutagenesis as an antiviral approach requires further investigation.
Purpose of the Study:
- To investigate the efficacy of 5-fluorouracil (FU) as a lethal mutagen for eliminating LCMV.
- To compare FU-mediated viral extinction with ribavirin's antiviral mechanism.
- To assess the in vivo feasibility of lethal mutagenesis for LCMV infection.
Main Methods:
- Treatment of cultured cells and mice with FU during LCMV infection.
- Measurement of viral mutation frequencies, replication, and transcription.
- Assessment of LCMV minigenome expression.
- Evaluation of persistent LCMV infection in immunodeficient mice.
Main Results:
- FU treatment led to efficient LCMV extinction in cell culture without significant cell toxicity.
- FU significantly increased mutation frequencies in viral genes, with minimal impact on replication and transcription per cycle.
- Ribavirin inhibited viral RNA synthesis and minigenome expression but did not increase mutation frequencies.
- FU treatment prevented LCMV persistent infection in vivo in B and T cell-deficient mice.
Conclusions:
- Lethal mutagenesis using FU is an effective strategy against LCMV, a negative-strand RNA virus.
- This approach differs mechanistically from ribavirin's antiviral action.
- In vivo prevention of persistent LCMV infection suggests the potential of lethal mutagenesis as a novel antiviral therapy.
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