Molecular characterization of attenuated Junin virus strains.

Romanowski, Vı́ctor; Ghiringhelli, Pablo Daniel; Albariño, César G.; Piboul, Mariel

doi:10.1099/0022-1317-78-7-1605

Cited by 41 publications

(37 citation statements)

References 50 publications

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“…24,25 These comparisons were complicated by the long passage histories of the vaccine strain candid #1 with respect to its virulent parent human isolate XJ, and the use of strain MC2, an unrelated rodent isolate, in the sequence analysis. In contrast, our approach compares PIC virus variants of markedly different pathogenicities that derive from a single naturally attenuated isolate.…”

Section: Discussionmentioning

confidence: 99%

Sequence analysis of the small RNA segment of guinea pig-passaged Pichinde virus variants.

Zhang

Marriott

Aronson³

1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. The established animal model for Lassa fever is based on the new world arenavirus Pichinde (PIC). Natural isolates of PIC virus are attenuated in guinea pigs, but serial guinea pig passage renders them extremely virulent in that host. We have compared the nucleotide sequences of the small RNA segments of two attenuated, lowpassage variants of the PIC virus Munchique strain (CoAn 4763) and two virulent, high-passage derivatives. Missense mutations in the glycoprotein precursor (GPC) gene at codons GPC-119, GPC-140, and GPC-164 and the nucleoprotein gene (NP) codons NP-35 and NP-374 were most closely associated with virulence. Codon GPC-140 is predicted to represent a region of peak hydrophilicity of the glycoprotein 1 (GP1); it is conceivable that mutations at this site could influence virulence by altering B cell epitopes or virus attachment protein conformation.Of all the viral hemorrhagic fevers of humans, Lassa fever (caused by the arenavirus Lassa) is most amenable to pathogenesis studies because of the availability of a wellcharacterized, safe, and accurate animal model. This model uses the new world arenavirus Pichinde (PIC), 1 a member of the Tacaribe serogroup, which is non-pathogenic for humans. Serial passage of the Munchique isolate of PIC virus (CoAn 4763) in inbred guinea pigs dramatically increases the pathogenicity of the virus. 2 Guinea pigs infected with this adapted PIC virus develop fever, leukopenia, thrombocytopenia, platelet dysfunction, 3 and terminal vascular collapse 4 in the absence of significant hemorrhagic manifestations, resembling Lassa fever. 5 Like Lassa virus in humans, adapted PIC virus replicates in all extraneural tissues of guinea pigs, and induces minimal histopathologic changes in liver, spleen and other reticuloendothelial organs of the infected host. [6][7][8] In contrast, the parental, unpassaged virus is naturally attenuated, and causes a self-limited febrile illness with low mortality in inbred guinea pigs. 2 This system offers the opportunity to delineate molecular genetic determinants of virulence. The relatively short passage history of parent (attenuated) and passaged (virulent) PIC virus suggests that sequence analysis would uncover a relatively small number of mutations between the two; the disparate virulence phenotypes favors the correlation of pathogenicity with specific viral genetic alterations.Arenaviruses have bipartite, ambisense, single-stranded RNA genomes. The small (S) segment encodes the viral envelope glycoproteins and nucleoprotein. 9 The glycoprotein precursor (GPC) protein is translated from a virus sense mRNA derived from the 5Ј half of the S segment. The GPC polypeptide is post-translationally processed by host cell signal peptidase and furin-like proteases to yield GP1 and GP2. Biochemical studies suggest that tetramers of GP2, which have a transmembrane domain, form a coiled-coil stalk, atop which tetramers of the globular GP1 protein are bound by non-covalent interactions.9,10 The cytoplasmic tails of the GP2 molecules are believe...

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Section: Discussionmentioning

confidence: 99%

Sequence analysis of the small RNA segment of guinea pig-passaged Pichinde virus variants.

Zhang

Marriott

Aronson³

1999

The American Journal of Tropical Medicine and Hygiene

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“…This new role of GP-2 may open new avenues in developing vaccines against Lassa virus using a rational hypothesis-driven approach rather than laborious, although highly successful, methods like classical serial passages as in the case of the Argentine hemorrhagic fever vaccine strain (42,43).…”

Section: A)mentioning

confidence: 99%

Evolution of Recombinant Lymphocytic Choriomeningitis Virus/Lassa Virus In Vivo Highlights the Importance of the GPC Cytosolic Tail in Viral Fitness

Sommerstein¹,

Palma

Ölschläger

et al. 2014

J Virol

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A key characteristic of arenaviruses is their ability to establish persistent infection in their natural host. Different factors like host age, viral dose strain, and route of infection may contribute to the establishment of persistence. However, the molecular mechanisms governing persistence are not fully understood. Here, we describe gain-of-function mutations of lymphocytic choriomeningitis virus (LCMV) expressing Lassa virus (LASV) GP, which can prolong viremia in mice depending on the sequences in the GP-2 cytoplasmic tail. The initial mutant variant (rLCMV/LASV mut GP) carried a point mutation in the cytosolic tail of the LASV glycoprotein GP corresponding to a K461G substitution. Unlike what occurred with the original rLCMV/LASV wild-type (wt) GP, infection of C57BL/6 mice with the mutated recombinant virus led to a detectable viremia of 2 weeks' duration. Further replacement of the entire sequence of the cytosolic tail from LASV to LCMV GP resulted in increased viral titers and delayed clearance of the viruses. Biosynthesis and cell surface localization of LASV wt and mut GPs were comparable. IMPORTANCEStarting from an emerging virus in a wild-type mouse, we engineered a panel of chimeric Lassa/lymphocytic choriomeningitis viruses. Mutants carrying a viral envelope with the cytosolic tail from the closely related mouse-adapted LCMV were able to achieve a productive viral infection lasting up to 27 days in wild-type mice. Biochemical assays showed a comparable biosynthesis and cell surface localization of LASV wt and mut GPs. These recombinant chimeric viruses could allow the study of immune responses and antivirals targeting the LASV GP.T he prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) represents a powerful experimental model used to study the virus-host interaction of noncytopathic viruses and the role of T cells in clearing viral infections in mouse models (1). Infection with several existing strains results in different outcomes causing either transient, acute infection with virus-specific protective immunity or protracted, chronic infection with persisting viremia and immunosuppression (2). LCMV is an enveloped virus comprising two segments (S and L) of ambisense singlestranded RNA. The S RNA encodes the nucleoprotein (NP) and the envelope glycoprotein precursor (GPC) that is posttranslationally cleaved by signal peptidase and the cellular proprotein convertase SKI-1/S1P into the mature virion glycoprotein complex SSP/GP-1/GP-2 (GPs). The L RNA encodes the RNA-dependent RNA polymerase (L) and a small RING finger protein (Z). The GP mediates cell target attachment and fusion. NP and Z cover several functions, including inhibition of the innate immune response and viral particle budding, respectively. NP and L assemble with the two ambisense segments to form the ribonucleoprotein complexes (RNPs), which serve as the templates for transcription and replication. It has been shown that in LCMV the viral polymerase L and GP-1 of the glycoprotein are important determinants for the...

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“…1A). Previous reports have compared Cd1-specific mutants to virulent strains of JUNV (5,19,26), but the contribution of any individual substitution to the attenuated phenotype has not generally been evaluated. In the present study, we investigated the functional consequences of the substitutions that have occurred in the Cd1 glycoprotein (GP).…”

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confidence: 99%

Substitutions in the Glycoprotein (GP) of the Candid#1 Vaccine Strain of Junin Virus Increase Dependence on Human Transferrin Receptor 1 for Entry and Destabilize the Metastable Conformation of GP

Droniou-Bonzom

Reignier

Oldenburg

et al. 2011

J Virol

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Candid#1 (Cd1) is an attenuated vaccine strain of Junin virus, the causative agent of Argentine hemorrhagic fever. Although several substitutions are present in Cd1, their importance for attenuation has not been established. We functionally characterized the substitutions present in the Cd1 glycoprotein (GP) and identified F427I in the transmembrane domain of the GP2 subunit as reducing infectivity in a reconstituted viral system. We further showed that this phenotype derives from the destabilization of the GP metastable conformation. Lastly, we identified an increased dependence of Cd1 GP on human transferrin receptor type 1 (hTfR-1) for entry, which may affect the tropism of the attenuated strain in vivo.

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Molecular characterization of attenuated Junin virus strains.

Cited by 41 publications

References 50 publications

Sequence analysis of the small RNA segment of guinea pig-passaged Pichinde virus variants.

Sequence analysis of the small RNA segment of guinea pig-passaged Pichinde virus variants.

Evolution of Recombinant Lymphocytic Choriomeningitis Virus/Lassa Virus In Vivo Highlights the Importance of the GPC Cytosolic Tail in Viral Fitness

Substitutions in the Glycoprotein (GP) of the Candid#1 Vaccine Strain of Junin Virus Increase Dependence on Human Transferrin Receptor 1 for Entry and Destabilize the Metastable Conformation of GP

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