A Novel BSL-2 Lassa Virus Reverse Genetics System Modelling the Complete Viral Life Cycle

Overview

Journal Emerg Microbes Infect

Specialty Infectious Diseases

Date 2024 May 15

PMID 38747061

Authors

Xiaoyou Hu

Xu Bai

Fangling Tian

Yifan Xing

Yi Shi

Yimin Tong

Jin Zhong

Affiliations

Soon will be listed here.

Abstract

Lassa virus (LASV), a risk-group 4 pathogen, must be handled in biosafety level-4 (BSL-4) conditions, thereby limiting its research and antiviral development. Here, we developed a novel LASV reverse genetics system which, to our knowledge, is the first to study the complete LASV life cycle under BSL-2 conditions. Viral particles can be produced efficiently when LASV minigenomic RNA harbouring minimal viral -elements and reporter genes is transfected into a helper cell line stably expressing viral NP, GP, Z and L proteins. The resulting defective virions, named LASVmg, can propagate only in the helper cell line, providing a BSL-2 model to study the complete LASV life cycle. Using this model, we found that a previously reported cellular receptor α-dystroglycan is dispensable for LASVmg infection. Furthermore, we showed that ribavirin can inhibit LASVmg infection by inducing viral mutations. This new BSL-2 system should facilitate studying the LASV life cycle and screening antivirals.

Citing Articles

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References

Salam A, Cheng V, Edwards T, Olliaro P, Sterne J, Horby P . Time to reconsider the role of ribavirin in Lassa fever. PLoS Negl Trop Dis. 2021; 15(7):e0009522. PMC: 8266111. DOI: 10.1371/journal.pntd.0009522. View

Loureiro M, DAntuono A, Lopez N . Virus⁻Host Interactions Involved in Lassa Virus Entry and Genome Replication. Pathogens. 2019; 8(1). PMC: 6470645. DOI: 10.3390/pathogens8010017. View

Yun N, Seregin A, Walker D, Popov V, Walker A, Smith J . Mice lacking functional STAT1 are highly susceptible to lethal infection with Lassa virus. J Virol. 2013; 87(19):10908-11. PMC: 3807383. DOI: 10.1128/JVI.01433-13. View

Ju X, Zhu Y, Wang Y, Li J, Zhang J, Gong M . A novel cell culture system modeling the SARS-CoV-2 life cycle. PLoS Pathog. 2021; 17(3):e1009439. PMC: 7990224. DOI: 10.1371/journal.ppat.1009439. View

Carrillo-Bustamante P, Nguyen T, Oestereich L, Gunther S, Guedj J, Graw F . Determining Ribavirin's mechanism of action against Lassa virus infection. Sci Rep. 2017; 7(1):11693. PMC: 5601963. DOI: 10.1038/s41598-017-10198-0. View

Horlbeck M, Gilbert L, Villalta J, Adamson B, Pak R, Chen Y . Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation. Elife. 2016; 5. PMC: 5094855. DOI: 10.7554/eLife.19760. View

Graci J, Cameron C . Mechanisms of action of ribavirin against distinct viruses. Rev Med Virol. 2005; 16(1):37-48. PMC: 7169142. DOI: 10.1002/rmv.483. View

Gan T, Zhou D, Huang Y, Xiao S, Ma Z, Hu X . Development of a New Reverse Genetics System for Ebola Virus. mSphere. 2021; 6(3). PMC: 8103987. DOI: 10.1128/mSphere.00235-21. View

Cornu T, de la Torre J . RING finger Z protein of lymphocytic choriomeningitis virus (LCMV) inhibits transcription and RNA replication of an LCMV S-segment minigenome. J Virol. 2001; 75(19):9415-26. PMC: 114509. DOI: 10.1128/JVI.75.19.9415-9426.2001. View

10.

Hara Y, Kanagawa M, Kunz S, Yoshida-Moriguchi T, Satz J, Kobayashi Y . Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection. Proc Natl Acad Sci U S A. 2011; 108(42):17426-31. PMC: 3198351. DOI: 10.1073/pnas.1114836108. View

11.

Sironi M, Forni D, de la Torre J . Mammarenavirus Genetic Diversity and Its Biological Implications. Curr Top Microbiol Immunol. 2023; 439:265-303. DOI: 10.1007/978-3-031-15640-3_8. View

12.

Moraz M, Pythoud C, Turk R, Rothenberger S, Pasquato A, Campbell K . Cell entry of Lassa virus induces tyrosine phosphorylation of dystroglycan. Cell Microbiol. 2013; 15(5):689-700. PMC: 3805106. DOI: 10.1111/cmi.12078. View

13.

Ortiz-Riano E, Cheng B, de la Torre J, Martinez-Sobrido L . Arenavirus reverse genetics for vaccine development. J Gen Virol. 2013; 94(Pt 6):1175-1188. PMC: 3709625. DOI: 10.1099/vir.0.051102-0. View

14.

Cornu T, Feldmann H, de la Torre J . Cells expressing the RING finger Z protein are resistant to arenavirus infection. J Virol. 2004; 78(6):2979-83. PMC: 353761. DOI: 10.1128/jvi.78.6.2979-2983.2004. View

15.

Kainulainen M, Spengler J, Welch S, Coleman-McCray J, Harmon J, Klena J . Use of a Scalable Replicon-Particle Vaccine to Protect Against Lethal Lassa Virus Infection in the Guinea Pig Model. J Infect Dis. 2018; 217(12):1957-1966. PMC: 6086598. DOI: 10.1093/infdis/jiy123. View

16.

PEDERSEN I . Density gradient centrifugation studies on lymphocytic choriomeningitis virus and on viral ribonucleic acid. J Virol. 1970; 6(4):414-20. PMC: 376138. DOI: 10.1128/JVI.6.4.414-420.1970. View

17.

Cornu T, de la Torre J . Characterization of the arenavirus RING finger Z protein regions required for Z-mediated inhibition of viral RNA synthesis. J Virol. 2002; 76(13):6678-88. PMC: 136245. DOI: 10.1128/jvi.76.13.6678-6688.2002. View

18.

Kunz S, Sevilla N, Rojek J, Oldstone M . Use of alternative receptors different than alpha-dystroglycan by selected isolates of lymphocytic choriomeningitis virus. Virology. 2004; 325(2):432-45. DOI: 10.1016/j.virol.2004.05.009. View

19.

Lukashevich I, Lemeshko N, Stelmakh T, Golubev V . [Pathogenic Machupo and Lassa arenaviruses: the biochemical properties of virion RNA and proteins]. Vopr Virusol. 1987; 32(4):473-80. View

20.

Ding Q, Huang B, Lu J, Liu Y, Zhong J . Hepatitis C virus NS3/4A protease blocks IL-28 production. Eur J Immunol. 2012; 42(9):2374-82. DOI: 10.1002/eji.201242388. View