scholarly journals Machupo Virus Expressing GPC of the Candid#1 Vaccine Strain of Junin Virus Is Highly Attenuated and Immunogenic

2015 ◽  
Vol 90 (3) ◽  
pp. 1290-1297 ◽  
Author(s):  
Takaaki Koma ◽  
Michael Patterson ◽  
Cheng Huang ◽  
Alexey V. Seregin ◽  
Payal D. Maharaj ◽  
...  
Keyword(s):  
Mouse Model ◽  
Vaccine Strain ◽  
Causative Agent ◽  
Transmembrane Domain ◽  
Hemorrhagic Fever ◽  
Single Amino Acid ◽  
Junin Virus ◽  
Machupo Virus ◽  
Junín Virus

ABSTRACTMachupo virus (MACV) is the causative agent of Bolivian hemorrhagic fever. Our previous study demonstrated that a MACV strain with a single amino acid substitution (F438I) in the transmembrane domain of glycoprotein is attenuated but genetically unstable in mice. MACV is closely related to Junin virus (JUNV), the causative agent of Argentine hemorrhagic fever. Others and our group have identified the glycoprotein to be the major viral factor determining JUNV attenuation. In this study, we tested the compatibility of the glycoprotein of the Candid#1 live-attenuated vaccine strain of JUNV in MACV replication and its ability to attenuate MACVin vivo. Recombinant MACV with the Candid#1 glycoprotein (rMACV/Cd#1-GPC) exhibited growth properties similar to those of Candid#1 and was genetically stablein vitro. In a mouse model of lethal infection, rMACV/Cd#1-GPC was fully attenuated, more immunogenic than Candid#1, and fully protective against MACV infection. Therefore, the MACV strain expressing the glycoprotein of Candid#1 is safe, genetically stable, and highly protective against MACV infection in a mouse model.IMPORTANCECurrently, there are no FDA-approved vaccines and/or treatments for Bolivian hemorrhagic fever, which is a fatal human disease caused by MACV. The development of antiviral strategies to combat viral hemorrhagic fevers, including Bolivian hemorrhagic fever, is one of the top priorities of the Implementation Plan of the U.S. Department of Health and Human Services Public Health Emergency Medical Countermeasures Enterprise. Here, we demonstrate for the first time that MACV expressing glycoprotein of Candid#1 is a safe, genetically stable, highly immunogenic, and protective vaccine candidate against Bolivian hemorrhagic fever.

scholarly journals Novel Arenavirus Entry Inhibitors Discovered by Using a Minigenome Rescue System for High-Throughput Drug Screening

2015 ◽  
Vol 89 (16) ◽  
pp. 8428-8443 ◽  
Author(s):  
Jessica Y. Rathbun ◽  
Magali E. Droniou ◽  
Robert Damoiseaux ◽  
Kevin G. Haworth ◽  
Jill E. Henley ◽  
...  
Keyword(s):  
High Throughput ◽  
Causative Agent ◽  
High Throughput Screening ◽  
Hemorrhagic Fever ◽  
Human Pathogens ◽  
Junin Virus ◽  
Hemorrhagic Fevers ◽  
Argentine Hemorrhagic Fever ◽  
Rescue System ◽  
Junín Virus

ABSTRACTCertain members of theArenaviridaefamily are category A agents capable of causing severe hemorrhagic fevers in humans. Specific antiviral treatments do not exist, and the only commonly used drug, ribavirin, has limited efficacy and can cause severe side effects. The discovery and development of new antivirals are inhibited by the biohazardous nature of the viruses, making them a relatively poorly understood group of human pathogens. We therefore adapted a reverse-genetics minigenome (MG) rescue system based on Junin virus, the causative agent of Argentine hemorrhagic fever, for high-throughput screening (HTS). The MG rescue system recapitulates all stages of the virus life cycle and enables screening of small-molecule libraries under biosafety containment level 2 (BSL2) conditions. The HTS resulted in the identification of four candidate compounds with potent activity against a broad panel of arenaviruses, three of which were completely novel. The target for all 4 compounds was the stage of viral entry, which positions the compounds as potentially important leads for future development.IMPORTANCEThe arenavirus family includes several members that are highly pathogenic, causing acute viral hemorrhagic fevers with high mortality rates. No specific effective treatments exist, and although a vaccine is available for Junin virus, the causative agent of Argentine hemorrhagic fever, it is licensed for use only in areas where Argentine hemorrhagic fever is endemic. For these reasons, it is important to identify specific compounds that could be developed as antivirals against these deadly viruses.

scholarly journals The Ectodomain of Glycoprotein from the Candid#1 Vaccine Strain of Junin Virus Rendered Machupo Virus Partially Attenuated in Mice Lacking IFN-αβ/γ Receptor

2016 ◽  
Vol 10 (8) ◽  
pp. e0004969 ◽  
Author(s):  
Takaaki Koma ◽  
Cheng Huang ◽  
Judith F. Aronson ◽  
Aida G. Walker ◽  
Milagros Miller ◽  
...  
Keyword(s):  
Vaccine Strain ◽  
Junin Virus ◽  
Machupo Virus ◽  
Junín Virus

scholarly journals Machupo virus with mutations in the transmembrane domain and glycosylation sites is attenuated and immunogenic in animal models of Bolivian Hemorrhagic Fever

2021 ◽  
Author(s):  
Emily K Mantlo ◽  
Junki Maruyama ◽  
John T Manning ◽  
Timothy G Wanninger ◽  
Cheng Huang ◽  
...  
Keyword(s):  
Animal Models ◽  
Mouse Model ◽  
Guinea Pigs ◽  
Rational Design ◽  
Transmembrane Domain ◽  
Vaccine Development ◽  
Hemorrhagic Fever ◽  
Highly Pathogenic ◽  
Glycosylation Sites ◽  
Machupo Virus

AbstractSeveral highly pathogenic mammarenaviruses cause severe hemorrhagic and neurologic disease in humans, for which vaccines and antivirals are limited or unavailable. New World (NW) mammarenavirus Machupo virus (MACV) infection causes Bolivian hemorrhagic fever in humans. We previously reported that the disruption of specific N-linked glycan sites on the glycoprotein (GPC) partially attenuate MACV in an IFN-αβ/γ receptor knockout mouse model. However, some capability to induce neurological pathology still remained. Highly pathogenic Junin virus (JUNV) is another NW arenavirus closely related to MACV. A F427I substitution in the GPC transmembrane domain (TMD) rendered JUNV attenuated in a lethal mouse model after intracranial inoculation. In this study, we rationally designed and rescued a MACV containing mutations at two glycosylation sites and the corresponding F438I substitution in GPC TMD. The MACV mutant is fully attenuated in IFN-αβ/γ receptor knockout mice and outbred guinea pigs. Furthermore, inoculation with this mutant MACV fully protected guinea pigs from wild-type MACV lethal challenge. Lastly, we found the GPC TMD F438I substitution greatly impaired MACV growth in neuronal cell lines of mouse and human origins. Our results highlight the critical roles of the glycans and the TMD on the GPC in arenavirus virulence, which informs the rational design of potential vaccine candidates for highly pathogenic arenaviruses.ImportanceFor arenaviruses, the only vaccine available is the live-attenuated Candid#1 vaccine, a JUNV vaccine approved in Argentina. We and others have found that the glycans on GPC and the F427 residue in the GPC TMD are important for virulence of JUNV. Nevertheless, mutating either of them is not sufficient for full and stable attenuation of JUNV. Using reverse genetics, we disrupted specific glycosylation sites on MACV GPC, and also introduced the corresponding F438I substitution in the GPC TMD. This MACV mutant is fully attenuated in two animal models and protects animals from lethal infection. Thus, our studies highlight the feasibility of rational attenuation of highly pathogenic arenaviruses for vaccine development. Another important finding from this study is that the F438I substitution in GPC TMD could substantially affect MACV replication in neurons. Future studies are warranted to elucidate the underlying mechanism and the implication of this mutation in arenavirus neural tropism.

Junin virus activity in two rural populations of the Argentine hemorrhagic fever (AHF) endemic area

1983 ◽  
Vol 12 (4) ◽  
pp. 273-280 ◽  
Author(s):  
Mercedes C. Weissenbacher ◽  
Marta S. Sabattini ◽  
María M. Avila ◽  
Patricia M. Sangiorgio ◽  
María R. F. De Sensi ◽  
...  
Keyword(s):  
Endemic Area ◽  
Hemorrhagic Fever ◽  
Rural Populations ◽  
Junin Virus ◽  
Argentine Hemorrhagic Fever ◽  
Virus Activity ◽  
Junín Virus

scholarly journals Development of Peptide-Conjugated Morpholino Oligomers as Pan-Arenavirus Inhibitors

2011 ◽  
Vol 55 (10) ◽  
pp. 4631-4638 ◽  
Author(s):  
Benjamin W. Neuman ◽  
Lydia H. Bederka ◽  
David A. Stein ◽  
Joey P. C. Ting ◽  
Hong M. Moulton ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Treatment Options ◽  
Hemorrhagic Fever ◽  
Virus Genome ◽  
Lymphocytic Choriomeningitis ◽  
Content Type ◽  
Junin Virus ◽  
Pichinde Virus ◽  
Junín Virus

ABSTRACTMembers of theArenaviridaefamily are a threat to public health and can cause meningitis and hemorrhagic fever, and yet treatment options remain limited by a lack of effective antivirals. In this study, we found that peptide-conjugated phosphorodiamidate morpholino oligomers (PPMO) complementary to viral genomic RNA were effective in reducing arenavirus replication in cell cultures andin vivo. PPMO complementary to the Junín virus genome were designed to interfere with viral RNA synthesis or translation or both. However, only PPMO designed to potentially interfere with translation were effective in reducing virus replication. PPMO complementary to sequences that are highly conserved across the arenaviruses and located at the 5′ termini of both genomic segments were effective against Junín virus, Tacaribe virus, Pichinde virus, and lymphocytic choriomeningitis virus (LCMV)-infected cell cultures and suppressed viral titers in the livers of LCMV-infected mice. These results suggest that arenavirus 5′ genomic termini represent promising targets for pan-arenavirus antiviral therapeutic development.

scholarly journals Development of a Reverse Genetic System to Generate Recombinant Chimeric Tacaribe Virus that Expresses Junín Virus Glycoproteins

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 948
Author(s):  
Sabrina Foscaldi ◽  
María Eugenia Loureiro ◽  
Claudia Sepúlveda ◽  
Carlos Palacios ◽  
María Belén Forlenza ◽  
...  
Keyword(s):  
Hemorrhagic Fever ◽  
Genetic System ◽  
Reverse Genetic System ◽  
Dependent Manner ◽  
Reverse Genetic ◽  
Junin Virus ◽  
Tacaribe Virus ◽  
Growth Properties ◽  
Intracellular Expression ◽  
Junín Virus

Mammarenaviruses are enveloped and segmented negative-stranded RNA viruses that comprise several pathogenic members associated with severe human hemorrhagic fevers. Tacaribe virus (TCRV) is the prototype for the New World group of mammarenaviruses and is not only naturally attenuated but also phylogenetically and antigenically related to all South American pathogenic mammarenaviruses, particularly the Junín virus (JUNV), which is the etiological agent of Argentinian hemorrhagic fever (AHF). Moreover, since TCRV protects guinea pigs and non-human primates from lethal challenges with pathogenic strains of JUNV, it has already been considered as a potential live-attenuated virus vaccine candidate against AHF. Here, we report the development of a reverse genetic system that relies on T7 polymerase-driven intracellular expression of the complementary copy (antigenome) of both viral S and L RNA segments. Using this approach, we successfully recovered recombinant TCRV (rTCRV) that displayed growth properties resembling those of authentic TCRV. We also generated a chimeric recombinant TCRV expressing the JUNV glycoproteins, which propagated similarly to wild-type rTCRV. Moreover, a controlled modification within the S RNA 5′ non-coding terminal sequence diminished rTCRV propagation in a cell-type dependent manner, giving rise to new perspectives where the incorporation of additional attenuation markers could contribute to develop safe rTCRV-based vaccines against pathogenic mammarenaviruses.

Endothelial cell function alteration after Junin virus infection

2003 ◽  
Vol 90 (08) ◽  
pp. 326-334 ◽  
Author(s):  
Ricardo Gomez ◽  
Roberto Pozner ◽  
Maria Lazzari ◽  
Lina D’Atri ◽  
Soledad Negrotto ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cell ◽  
Cell Function ◽  
Umbilical Vein ◽  
Hemorrhagic Fever ◽  
No Production ◽  
Epidemic Disease ◽  
Endothelial Cell Function ◽  
Junin Virus ◽  
Junín Virus

SummaryHematologic involvement is the main feature of Argentine hemorrhagic fever (AHF), an endemo-epidemic disease caused by Junin virus (JV). Since endothelial dysfunction could play a role in AHF-altered hemostasis, we studied human umbilical vein endothelial cell (HUVEC) infection with a virulent (JVv) and a non-virulent (JVa) JV strain. Cells were infected by the two JV variants with no detectable apoptosis or cytopathic effect. Both viral variants up-regulated ICAM-1 and VCAM-1 levels, while von Willebrand factor (VWF) production was decreased. Prostacyclin (PGI2) release and decay accelerating factor (DAF) expression were greater in JVv- than in JVa-infected or control cells. Furthermore, nitric oxide (NO) production and endothelial nitric oxide synthase (eNOS) expression was only raised in JVv-infected supernatants. Significant NO and PGI2 values were also detected in AHF patient sera.These data demonstrate that endothelial cell responses are triggered subsequently by JV infection, suggesting that such alterations play a major role in the pathogenesis of AHF and perhaps in other viral-induced hemorrhagic diseases.

scholarly journals Junín Virus Promotes Autophagy To Facilitate the Virus Life Cycle

2019 ◽  
Vol 93 (15) ◽  
Author(s):  
Julieta S. Roldán ◽  
Nélida A. Candurra ◽  
María I. Colombo ◽  
Laura R. Delgui
Keyword(s):  
Hemorrhagic Fever ◽  
Replication Capacity ◽  
Epidemic Disease ◽  
Junin Virus ◽  
Hemorrhagic Fevers ◽  
Argentine Hemorrhagic Fever ◽  
The Family ◽  
Infected Cells ◽  
Junín Virus

ABSTRACTJunín virus (JUNV), a member of the familyArenaviridae, is the etiological agent of Argentine hemorrhagic fever (AHF), a potentially deadly endemic-epidemic disease affecting the population of the most fertile farming land of Argentina. Autophagy is a degradative process with a crucial antiviral role; however, several viruses subvert the pathway to their benefit. We determined the role of autophagy in JUNV-infected cells by analyzing LC3, a cytoplasmic protein (LC3-I) that becomes vesicle membrane associated (LC3-II) upon induction of autophagy. Cells overexpressing enhanced green fluorescent protein (EGFP)-LC3 and infected with JUNV showed an increased number of LC3 punctate structures, similar to those obtained after starvation or bafilomycin A1 treatment, which leads to autophagosome induction or accumulation, respectively. We also monitored the conversion of LC3-I to LC3-II, observing LC3-II levels in JUNV-infected cells similar to those observed in starved cells. Additionally, we kinetically studied the number of LC3 dots after JUNV infection and found that the virus activated the pathway as early as 2 h postinfection (p.i.), whereas the UV-inactivated virus did not induce the pathway. Cells subjected to starvation or pretreated with rapamycin, a pharmacological autophagy inductor, enhanced virus yield. Also, we assayed the replication capacity of JUNV in Atg5 knockout or Beclin 1 knockdown cells (both critical components of the autophagic pathway) and found a significant decrease in JUNV replication. Taken together, our results constitute the first study indicating that JUNV infection induces an autophagic response, which is functionally required by the virus for efficient propagation.IMPORTANCEMammalian arenaviruses are zoonotic viruses that cause asymptomatic and persistent infections in their rodent hosts but may produce severe and lethal hemorrhagic fevers in humans. Currently, there are neither effective therapeutic options nor effective vaccines for viral hemorrhagic fevers caused by human-pathogenic arenaviruses, except the vaccine Candid no. 1 against Argentine hemorrhagic fever (AHF), licensed for human use in areas of endemicity in Argentina. Since arenaviruses remain a severe threat to global public health, more in-depth knowledge of their replication mechanisms would improve our ability to fight these viruses. Autophagy is a lysosomal degradative pathway involved in maintaining cellular homeostasis, representing powerful anti-infective machinery. We show, for the first time for a member of the familyArenaviridae, a proviral role of autophagy in JUNV infection, providing new knowledge in the field of host-virus interaction. Therefore, modulation of virus-induced autophagy could be used as a strategy to block arenavirus infections.

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