TY - JOUR
T1 - Lassa virus vaccine candidate ml29 generates truncated viral rnas which contribute to interfering activity and attenuation
AU - Johnson, Dylan M.
AU - Cubitt, Beatrice
AU - Pfeffer, Tia L.
AU - de la Torre, Juan Carlos
AU - Lukashevich, Igor S.
N1 - Funding Information:
This research was funded by the NIH National Institute of Allergy and Infectious Disease, grant number 1R56AI135770-01A1. The content of this manuscript is solely the responsibility of the authors and does not necessarily represent the official views of the funding agency. We would like to thank Slobodan Paessler and John Tyler Manning (University of Texas Medical Branch, Galveston, TX, USA) for providing LASV RNA for Northern blotting, and for assisting with the replication of startle response measurements to ensure identical conditions were used in our experiments to those used in the previously published work characterizing sensorineural hearing loss in LASV infected mice. We would also like to thank Kenneth Palmer and Divyasha Saxena (University of Louisville, Louisville, KY, USA) for their assistance with the LCMV-WE guinea pig control experiments.
Funding Information:
Funding: This research was funded by the NIH National Institute of Allergy and Infectious Disease, grant number 1R56AI135770-01A1. The content of this manuscript is solely the responsibility of the authors and does not necessarily represent the official views of the funding agency.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/2
Y1 - 2021/2
N2 - Defective interfering particles (DIPs) are naturally occurring products during virus replication in infected cells. DIPs contain defective viral genomes (DVGs) and interfere with replication and propagation of their corresponding standard viral genomes by competing for viral and cellular resources, as well as promoting innate immune antiviral responses. Consequently, for many different viruses, including mammarenaviruses, DIPs play key roles in the outcome of infection. Due to their ability to broadly interfere with viral replication, DIPs are attractive tools for the development of a new generation of biologics to target genetically diverse and rapidly evolving viruses. Here, we provide evidence that in cells infected with the Lassa fever (LF) vaccine candidate ML29, a reassortant that carries the nucleoprotein (NP) and glycoprotein (GP) dominant antigens of the pathogenic Lassa virus (LASV) together with the L polymerase and Z matrix protein of the non-pathogenic genetically related Mopeia virus (MOPV), L-derived truncated RNA species are readily detected following infection at low multiplicity of infection (MOI) or in persistently-infected cells originally infected at high MOI. In the present study, we show that expression of green fluorescent protein (GFP) driven by a tri-segmented form of the mammarenavirus lymphocytic choriomeningitis virus (r3LCMV-GFP/GFP) was strongly inhibited in ML29-persistently infected cells, and that the magnitude of GFP suppression was dependent on the passage history of the ML29-persistently infected cells. In addition, we found that DIP-enriched ML29 was highly attenuated in immunocompetent CBA/J mice and in Hartley guinea pigs. Likewise, STAT-1-/- mice, a validated small animal model for human LF associated hearing loss sequelae, infected with DIP-enriched ML29 did not exhibit any hearing abnormalities throughout the observation period (62 days).
AB - Defective interfering particles (DIPs) are naturally occurring products during virus replication in infected cells. DIPs contain defective viral genomes (DVGs) and interfere with replication and propagation of their corresponding standard viral genomes by competing for viral and cellular resources, as well as promoting innate immune antiviral responses. Consequently, for many different viruses, including mammarenaviruses, DIPs play key roles in the outcome of infection. Due to their ability to broadly interfere with viral replication, DIPs are attractive tools for the development of a new generation of biologics to target genetically diverse and rapidly evolving viruses. Here, we provide evidence that in cells infected with the Lassa fever (LF) vaccine candidate ML29, a reassortant that carries the nucleoprotein (NP) and glycoprotein (GP) dominant antigens of the pathogenic Lassa virus (LASV) together with the L polymerase and Z matrix protein of the non-pathogenic genetically related Mopeia virus (MOPV), L-derived truncated RNA species are readily detected following infection at low multiplicity of infection (MOI) or in persistently-infected cells originally infected at high MOI. In the present study, we show that expression of green fluorescent protein (GFP) driven by a tri-segmented form of the mammarenavirus lymphocytic choriomeningitis virus (r3LCMV-GFP/GFP) was strongly inhibited in ML29-persistently infected cells, and that the magnitude of GFP suppression was dependent on the passage history of the ML29-persistently infected cells. In addition, we found that DIP-enriched ML29 was highly attenuated in immunocompetent CBA/J mice and in Hartley guinea pigs. Likewise, STAT-1-/- mice, a validated small animal model for human LF associated hearing loss sequelae, infected with DIP-enriched ML29 did not exhibit any hearing abnormalities throughout the observation period (62 days).
KW - Defective interfering particles
KW - Homologous and heterologous interference
KW - LASV vaccine development
KW - Lassa virus (LASV)
KW - Molecular virology of mammalian arenaviruses
KW - Safety in small animal models
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U2 - 10.3390/v13020214
DO - 10.3390/v13020214
M3 - Article
C2 - 33573250
AN - SCOPUS:85101440756
VL - 13
JO - Viruses
JF - Viruses
SN - 1999-4915
IS - 2
M1 - 214
ER -