# ent

id: cord-001765-7wv4cb37
type: WORK_OF_ART
entity:
  Vaccination With a Highly Attenuated Recombinant Vesicular Stomatitis Virus Vector Protects Against Challenge With a Lethal Dose of Ebola Virus


id: cord-004126-u6ts87ur
type: ORG
entity:
  the Integrated Research Facility Rocky Mountain Laboratories RML Division of Intramural Research DIR


id: cord-004126-u6ts87ur
type: ORG
entity:
  National Institute of Allergy and Infectious Disease NIAID National Institutes of Health NIH Vaccinations


id: cord-000660-tsvzg0ax
type: DNA
entity:
  NCBI Entrez Nucleotide database Ifit2 NM_008332 Ifit1 NM_008331 Ifit3 NM_010501 Ifnb1 NM_010510 Ifnar1 NM_010508 Figure S1 Survival of wt and Ifit2 2 2


id: cord-000660-tsvzg0ax
type: CHEMICAL
entity:
  IFN alpha beta gene induction Essential role of IPS 1 in innate immune responses against RNA viruses


id: cord-000660-tsvzg0ax
type: CHEMICAL
entity:
  interferon alpha beta or gamma using oligonucleotide arrays Interferon inducible antiviral effectors A


id: cord-004733-i0a3igc7
type: PROTEIN
entity:
  alkaline phosphatase conjugated goat anti mouse IgG M Tago 6553 Tago Inc Burlingame CA U S A diluted 5 000 fold


id: cord-003667-u1xa44nw
type: ORG
entity:
  Properties of Replication Competent Vesicular Stomatitis Virus Vectors Expressing Glycoproteins of Filoviruses and Arenaviruses Properties of Replication Competent Vesicular Stomatitis Virus Vectors Expressing Glycoproteins of Filoviruses


id: cord-262753-jld1ygxt
type: PROTEIN
entity:
  primary antibodies rabbit anti eIF3a Cell Signaling 3411 rabbit anti eIF4E Cell Signaling 9742 rabbit anti 4E BP1 Cell Signaling 9452 rabbit anti phospho 4E BP1 Ser65 Cell Signaling 9451 rabbit anti phospho 4E BP1 Thr37 46 Cell Signaling 2855 mouse anti actin Millipore MAB1501 mouse anti actin


id: cord-262753-jld1ygxt
type: DISEASE
entity:
  influenza A virus a segmented negative strand RNA virus that replicates in the nucleus 50 of the total mRNA in the cytoplasm is viral 48 In this case however the viral endonuclease PA X degrades cellular mRNA which further contributes to the dominance of viral mRNA 66 67 For vaccinia virus a DNA virus that replicates entirely within the cytoplasm degradation


id: cord-020714-h1fevqcw
type: PROTEIN
entity:
  neuraminidase inhibitor 2 deoxy 2 3 dehydro Ntrifluoracetylneuraminic acid FANA Influenza virus grown


id: cord-020714-h1fevqcw
type: DISEASE
entity:
  herpes simplex virus XII The virion polypeptides of type 1 strains Protein composition of coronavirus OC


id: cord-020714-h1fevqcw
type: DISEASE
entity:
  herpes simplex virus IV The site of glycosylation and accumulation of viral membrane proteins The polypeptides of influenza virus VII


id: cord-267712-mhx8e5y0
type: DISEASE
entity:
  Influenza virus Ebola virus Marburg virus Human immunodeficiency HIV virus Severe Acute Respiratory Syndrome SARS virus and Hepatitis C virus 5 6 7 8 9


id: cord-272051-arz8r204
type: WORK_OF_ART
entity:
  A R Castrucci and A M Ciccaglione Department of Infectious Parasitic and Immunomediated Diseases Istituto Superiore


id: cord-011435-x73foqu7
type: DNA
entity:
  R C and R T T visualization S C A G K C S F and G S supervision S C R C and R T T project administration S C R C and R T T funding acquisition S C and R T T


id: cord-011435-x73foqu7
type: WORK_OF_ART
entity:
  Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High Fat Diet Induced Liver Injury


id: cord-000079-533xlisc
type: PROTEIN
entity:
  human specific mouse anti nestin antibodies Millipore Billerica MA goat anti SOX2 antibodies R D mouse anti b2tubulinIII Millipore antibodies


id: cord-000079-533xlisc
type: CARDINAL
entity:
  between 400 and 600 mm were selected for in vitro experiments and for intracerebral implantation Tumors were dissociated


id: cord-291323-kbjyd5g3
type: DISEASE
entity:
  influenza A virus and vesicular stomatitis virus VSV For Zaire ebolavirus ZEBOV proteolytic processing of the envelope


id: cord-104239-xxlcdbqi
type: CHEMICAL
entity:
  thank Dr G Palade Dr M G Farquhar and Michael McCaffery for their many helpful comments concerning the EM This work


id: cord-316589-f1hq0xl5
type: WORK_OF_ART
entity:
  Oncolytic Recombinant Vesicular Stomatitis Virus VSV Is Nonpathogenic and Nontransmissible in Pigs a Natural Host of VSV Safety Studies in Tumor and Non Tumor


id: cord-316589-f1hq0xl5
type: ORG
entity:
  Carriers Comparative Oncology Evaluation of Intravenous Recombinant Oncolytic Vesicular Stomatitis Virus Therapy


id: cord-326013-5i35zdmv
type: PROTEIN
entity:
  cathepsin D 14 phospholipase A 2 15 ceramide activated protein serine threonine phosphatases CAPP 16 protein kinase C isoforms 17 18


id: cord-296466-hakaoo9i
type: DNA
entity:
  baculovirus Autographa californica nuclear polyhedrosis virus A novel method for viral display of ER membrane proteins


id: cord-296466-hakaoo9i
type: CHEMICAL
entity:
  Autographa californica multicapsid nucleopolyhedrovirus The baculovirus GP64 envelope fusion protein Synthesis oligomerization and processing A


id: cord-296187-nnv2e7gr
type: DNA
entity:
  EcoRI and BglII sites pVSV eGFP dG 31842 pMD2 G 12259 pCAG VSV P 64088 pCAG VSV L 64085 pCAG VSV N 64087


id: cord-299281-5z1xminb
type: CHEMICAL
entity:
  8 1 4 galactosyltransferase specifies trans Golgi localization Overlapping distribution of two glycosyltransferases in the Golgi apparatus of


id: cord-290243-m8yfugr0
type: DISEASE
entity:
  HIV human immunodeficiency virus type 1 and 2 EMCV encephalomyocarditis virus Cox B3 Coxsackie B virus type 3 VSV vesicular stomatitis virus and HSV herpes simplex virus type 1 and 2 Table 1 EC


id: cord-290243-m8yfugr0
type: CHEMICAL
entity:
  Benzoic acid 3S 4S 5R 4 benzyloxy 5 tert butyl diphenyl silanyloxymethyl tetrahydro thiophen 3 yl ester


id: cord-346554-a98pjtxs
type: CELL_LINE
entity:
  CCL 2TM Ceropithecus aethiops epithelial kidney Vero ATCC CCL 81 human epithelial HeLa contaminant carcinoma HEp 2


id: cord-276009-p98wjtjb
type: DNA
entity:
  5 and 3 using 5 WNE FLAG Not I 5 GACGACGCGGCCGC ATGTTTAACTGCCTTGGAA TGAGC 3 and 3 WNE FLAG BamHI 5 GCAGCAGGATCCAGCGTGCACGTTCACGG AGAGG 3 primers NotI


id: cord-276009-p98wjtjb
type: DNA
entity:
  5 XN2 Xho I 5 CCGCGGCTCGAGATGTTT AACTGCCTTGGAATGAGC 3 and 3 XN2 Nhe I 5 GACGACGCTAGCGGATCACTAC TTGTCATCGTC 3 primers XhoI


id: cord-296399-vvbjulm9
type: GGP
entity:
  tetherin s N glycosylation status might contribute to this effect Moreover our results indicate that findings made for tetherin


id: cord-296399-vvbjulm9
type: WORK_OF_ART
entity:
  The Tetherin Antagonism of the Ebola Virus Glycoprotein Requires an Intact Receptor Binding Domain and Can Be Blocked by GP1 Specific Antibodies Fusionactive glycoprotein G


id: cord-296399-vvbjulm9
type: WORK_OF_ART
entity:
  The Glycoproteins of All Filovirus Species Use the Same Host Factors for Entry into Bat and Human Cells but Entry Efficiency


id: cord-351881-qea4b0i5
type: DNA
entity:
  Olot 91 strain GenBank Accession Number AGW23409 was synthesized by GenScript Piscataway For generation


id: cord-327199-ggomuomb
type: CHEMICAL
entity:
  Autographa californica nucleopolyhedrovirus budded virions Correlation between structure protein composition morphogenesis and cytopathology of Glossina pallidipes salivary gland hypertrophy


id: cord-327199-ggomuomb
type: CHEMICAL
entity:
  alphavirus budding Influenza virus assembly and budding in raft derived microdomains a quantitative analysis of the surface distribution of HA NA


# bib

id: cord-000079-533xlisc
author: Huszthy, Peter C.
date: 2009-07-20
txt: ./txt/cord-000079-533xlisc.txt
cache: ./cache/cord-000079-533xlisc.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2707627/
doi: 10.1371/journal.pone.0006314
summary:
  Both, lymphocytic choriomeningitis virus glycoprotein (LCMV-GP) and vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviral vectors very efficiently transduced human glioblastoma cells in vitro and in vivo. In a therapeutic approach using the suicide gene herpes simplex virus thymidine kinase (HSV-1-tk) fused to eGFP, both lentiviral vectors mediated a complete remission of solid tumors as seen on MRI resulting in a highly significant survival benefit (p<0.001) compared to control groups. Furthermore, we showed a significant therapeutic effect of LCMV-GP pseudotyped lentiviral vectors in the cell-line based 9L rat glioma model using the suicide gene HSV-1-tk. In the presented work, we showed that both, VSV-G and LCMV-GP pseudotyped lentiviruses efficiently transduced human glioma cells in vitro and in vivo, whereas gammaretroviral transduction was inefficient. When analyzed at higher magnification, both LCMV-GP and VSV-G pseudotyped lentiviral vectors showed efficient transgene delivery to nestin-positive tumor cells in solid ( Figure 3B ,E) and invasive tumor areas ( Figure 3C ,F).

title:
  Remission of Invasive, Cancer Stem-Like Glioblastoma Xenografts Using Lentiviral Vector-Mediated Suicide Gene Therapy

abstract:
  BACKGROUND: Glioblastoma is the most frequent and most malignant primary brain tumor with a poor prognosis. The translation of therapeutic strategies for glioblastoma from the experimental phase into the clinic has been limited by insufficient animal models, which lack important features of human tumors. Lentiviral gene therapy is an attractive therapeutic option for human glioblastoma, which we validated in a clinically relevant animal model. METHODOLOGY/PRINCIPAL FINDINGS: We used a rodent xenograft model that recapitulates the invasive and angiogenic features of human glioblastoma to analyze the transduction pattern and therapeutic efficacy of lentiviral pseudotyped vectors. Both, lymphocytic choriomeningitis virus glycoprotein (LCMV-GP) and vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviral vectors very efficiently transduced human glioblastoma cells in vitro and in vivo. In contrast, pseudotyped gammaretroviral vectors, similar to those evaluated for clinical therapy of glioblastoma, showed inefficient gene transfer in vitro and in vivo. Both pseudotyped lentiviral vectors transduced cancer stem-like cells characterized by their CD133-, nestin- and SOX2-expression, the ability to form spheroids in neural stem cell medium and to express astrocytic and neuronal differentiation markers under serum conditions. In a therapeutic approach using the suicide gene herpes simplex virus thymidine kinase (HSV-1-tk) fused to eGFP, both lentiviral vectors mediated a complete remission of solid tumors as seen on MRI resulting in a highly significant survival benefit (p<0.001) compared to control groups. In all recurrent tumors, surviving eGFP-positive tumor cells were found, advocating prodrug application for several cycles to even enhance and prolong the therapeutic effect. CONCLUSIONS/SIGNIFICANCE: In conclusion, lentiviral pseudotyped vectors are promising candidates for gene therapy of glioma in patients. The inefficient gene delivery by gammaretroviral vectors is in line with the results obtained in clinical therapy for GBM and thus confirms the high reproducibility of the invasive glioma animal model for translational research.


id: cord-000660-tsvzg0ax
title: Interferon-Induced Ifit2/ISG54 Protects Mice from Lethal VSV Neuropathogenesis
author: Fensterl, Volker
date: 2012-05-17
txt: ./txt/cord-000660-tsvzg0ax.txt
cache: ./cache/cord-000660-tsvzg0ax.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3355090/
doi: 10.1371/journal.ppat.1002712
summary:
  Interferon protects mice from vesicular stomatitis virus (VSV) infection and pathogenesis; however, it is not known which of the numerous interferon-stimulated genes (ISG) mediate the antiviral effect. In contrast, in acute infection by viruses that cause severe pathogenesis and death within a few days after infection, protection is primarily provided by the intrinsic antiviral actions of IFN-induced proteins encoded by the hundreds of IFN-stimulated genes (ISGs) [10] [11] [12] , several of which often contribute to the overall effect of IFN against a given virus. From the above observations, we conclude that after intranasal infection by VSV, Ifit2 protects mice from neuropathogenesis by suppressing replication or spread of the virus in brain neurons. In the complete absence of type I IFN action in the IFNAR 2/2 mice, intranasally infected VSV replicated vigorously not only in brains, but also in livers and lungs ( Figure 7A-C) .

abstract:
  Interferon protects mice from vesicular stomatitis virus (VSV) infection and pathogenesis; however, it is not known which of the numerous interferon-stimulated genes (ISG) mediate the antiviral effect. A prominent family of ISGs is the interferon-induced with tetratricopeptide repeats (Ifit) genes comprising three members in mice, Ifit1/ISG56, Ifit2/ISG54 and Ifit3/ISG49. Intranasal infection with a low dose of VSV is not lethal to wild-type mice and all three Ifit genes are induced in the central nervous system of the infected mice. We tested their potential contributions to the observed protection of wild-type mice from VSV pathogenesis, by taking advantage of the newly generated knockout mice lacking either Ifit2 or Ifit1. We observed that in Ifit2 knockout (Ifit2 (−/−)) mice, intranasal VSV infection was uniformly lethal and death was preceded by neurological signs, such as ataxia and hind limb paralysis. In contrast, wild-type and Ifit1 (−/−) mice were highly protected and survived without developing such disease. However, when VSV was injected intracranially, virus replication and survival were not significantly different between wild-type and Ifit2(−/−) mice. When administered intranasally, VSV entered the central nervous system through the olfactory bulbs, where it replicated equivalently in wild-type and Ifit2 (−/−) mice and induced interferon-β. However, as the infection spread to other regions of the brain, VSV titers rose several hundred folds higher in Ifit2 (−/−) mice as compared to wild-type mice. This was not caused by a broadened cell tropism in the brains of Ifit2 (−/−) mice, where VSV still replicated selectively in neurons. Surprisingly, this advantage for VSV replication in the brains of Ifit2(−/−) mice was not observed in other organs, such as lung and liver. Pathogenesis by another neurotropic RNA virus, encephalomyocarditis virus, was not enhanced in the brains of Ifit2 (−/−) mice. Our study provides a clear demonstration of tissue-, virus- and ISG-specific antiviral action of interferon.


id: cord-001765-7wv4cb37
author: Matassov, Demetrius
date: 2015-06-24
txt: ./txt/cord-001765-7wv4cb37.txt
cache: ./cache/cord-001765-7wv4cb37.txt
extension: .txt
mime: text/plain
url: http://europepmc.org/articles/pmc4564554?pdf=render
doi: 10.1093/infdis/jiv316
summary:
  One of these rVSV vectors (N4CT1-EBOVGP1), which expresses membrane-anchored EBOV GP from the first position in the genome (GP1), elicited a balanced cellular and humoral GP-specific immune response in mice. Guinea pigs immunized with a single dose of this vector were protected from any signs of disease following lethal EBOV challenge, while control animals died in 7-9 days. Guinea pigs immunized with a single dose of this vector were protected from any signs of disease following lethal EBOV challenge, while control animals died in 7-9 days. The studies described here are the first to demonstrate protection of guinea pigs and macaques with a single dose of highly attenuated rVSV expressing EBOVGP, and we believe that the N4CT1-EBOVGP1 vector has the essential safety and efficacy characteristics for use in a vaccine to prevent EBOV infection in humans and the great apes.

title:
  Vaccination With a Highly Attenuated Recombinant Vesicular Stomatitis Virus Vector Protects Against Challenge With a Lethal Dose of Ebola Virus

abstract:
  Previously, recombinant vesicular stomatitis virus (rVSV) pseudotypes expressing Ebolavirus glycoproteins (GPs) in place of the VSV G protein demonstrated protection of nonhuman primates from lethal homologous Ebolavirus challenge. Those pseudotype vectors contained no additional attenuating mutations in the rVSV genome. Here we describe rVSV vectors containing a full complement of VSV genes and expressing the Ebola virus (EBOV) GP from an additional transcription unit. These rVSV vectors contain the same combination of attenuating mutations used previously in the clinical development pathway of an rVSV/human immunodeficiency virus type 1 vaccine. One of these rVSV vectors (N4CT1-EBOVGP1), which expresses membrane-anchored EBOV GP from the first position in the genome (GP1), elicited a balanced cellular and humoral GP-specific immune response in mice. Guinea pigs immunized with a single dose of this vector were protected from any signs of disease following lethal EBOV challenge, while control animals died in 7–9 days. Subsequently, N4CT1-EBOVGP1 demonstrated complete, single-dose protection of 2 macaques following lethal EBOV challenge. A single sham-vaccinated macaque died from disease due to EBOV infection. These results demonstrate that highly attenuated rVSV vectors expressing EBOV GP may provide safer alternatives to current EBOV vaccines.


id: cord-003667-u1xa44nw
title: Vesicular Stomatitis Virus-Based Vaccine Protects Mice against Crimean-Congo Hemorrhagic Fever
author: Rodriguez, Sergio E.
date: 2019-05-23
txt: ./txt/cord-003667-u1xa44nw.txt
cache: ./cache/cord-003667-u1xa44nw.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6533279/
doi: 10.1038/s41598-019-44210-6
summary:
  Based on the results of these initial pilot studies, we elected to adjust vaccine and challenge doses, and administered 10 7 pfu/dose of the replication competent virus (ΔGrVSV-CCHFV-GPCΔ) to prime and boosted groups of five STAT-1 −/− mice, respectively (Fig. 3) . Regardless, protection was achieved by both regimens, although the boosted group data suggests that at study endpoint, the observed IgG titers against CCHFV-GPC along with lower neutralizing titers (PRNT 50 of < 1:320) are evidence of the ability to combat lethal CCHFV infection in the STAT-1 −/− mouse model after vaccination (Fig. 5A,B) . Now that we have established that this new vector can provide protective benefit, our future studies will temporally examine the antibody and T cell repertoire after prime and boosting doses following ΔGrVSV-CCHFV-GPCΔ, but before CCHFV challenge, as these would be informative for the STAT-1 −/− mouse model.

abstract:
  Crimean-Congo hemorrhagic fever virus (CCHFV), a tick-borne bunyavirus, can cause a life-threatening hemorrhagic syndrome in humans but not in its animal host. The virus is widely distributed throughout southeastern Europe, the Middle East, Africa, and Asia. Disease management has proven difficult and there are no broadly licensed vaccines or therapeutics. Recombinant vesicular stomatitis viruses (rVSV) expressing foreign glycoproteins (GP) have shown promise as experimental vaccines for several viral hemorrhagic fevers. Here, we developed and assessed a replication competent rVSV vector expressing the CCHFV glycoprotein precursor (GPC), which encodes CCHFV structural glycoproteins. This construct drives strong expression of CCHFV-GP, in vitro. Using these vectors, we vaccinated STAT-1 knock-out mice, an animal model for CCHFV. The vector was tolerated and 100% efficacious against challenge from a clinical strain of CCHFV. Anti-CCHFV-GP IgG and neutralizing antibody titers were observed in surviving animals. This study demonstrates that a rVSV expressing only the CCHFV-GP has the potential to serve as a replication competent vaccine platform against CCHF infections.


id: cord-004126-u6ts87ur
author: Furuyama, Wakako
date: 2020-01-10
txt: ./txt/cord-004126-u6ts87ur.txt
cache: ./cache/cord-004126-u6ts87ur.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6954110/
doi: 10.1038/s41541-019-0155-z
summary:
  Moreover, single vaccination induced cross-protective H5-specific antibodies and protected mice against lethal challenge with various H5 clade 2 viruses, highlighting the potential of the VSV-based HAfl as a pan-H5 influenza virus emergency vaccine. We found that a single vaccination with VSV-vectors expressing the full-length HA (HAfl) induced crossreactive H5-specific antibodies and conferred complete protection against lethal challenge with various H5 clade 2 viruses. In contrast to all the sHA-based vaccines, single doses of the VSV-EBOV-HAfl or VSV-HAfl vectors were sufficient to provide complete protection from lethal homologous H5N1 challenge in mice (Fig. 2) . However, this study did not provide any data supporting an advantage of including VSV-EBOV as part of the vector design over just expressing VSV-HAfl as both vectors performed similarly well with no statistically significant difference in efficacy following single-dose or prime/boost administration (Fig. 2 ) nor in antibody responses (Fig. 3, Table 1 ).

title:
  A single dose of a vesicular stomatitis virus-based influenza vaccine confers rapid protection against H5 viruses from different clades

abstract:
  The avian influenza virus outbreak in 1997 highlighted the potential of the highly pathogenic H5N1 virus to cause severe disease in humans. Therefore, effective vaccines against H5N1 viruses are needed to counter the potential threat of a global pandemic. We have previously developed a fast-acting and efficacious vaccine against Ebola virus (EBOV) using the vesicular stomatitis virus (VSV) platform. In this study, we generated recombinant VSV-based H5N1 influenza virus vectors to demonstrate the feasibility of this platform for a fast-acting pan-H5 influenza virus vaccine. We chose multiple approaches regarding antigen design and genome location to define a more optimized vaccine approach. After the VSV-based H5N1 influenza virus constructs were recovered and characterized in vitro, mice were vaccinated by a single dose or prime/boost regimen followed by challenge with a lethal dose of the homologous H5 clade 1 virus. We found that a single dose of VSV vectors expressing full-length hemagglutinin (HAfl) were sufficient to provide 100% protection. The vaccine vectors were fast-acting as demonstrated by uniform protection when administered 3 days prior to lethal challenge. Moreover, single vaccination induced cross-protective H5-specific antibodies and protected mice against lethal challenge with various H5 clade 2 viruses, highlighting the potential of the VSV-based HAfl as a pan-H5 influenza virus emergency vaccine.


id: cord-004733-i0a3igc7
author: Nagata, S.
date: 1992
txt: ./txt/cord-004733-i0a3igc7.txt
cache: ./cache/cord-004733-i0a3igc7.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7086791/
doi: 10.1007/bf01309581
summary:
  title: Identification of epitopes associated with different biological activities on the glycoprotein of vesicular stomatitis virus by use of monoclonal antibodies Thirteen monoclonal antibodies (MAbs) to the glycoprotein (G) of vesicular stomatitis virus (VSV) serotype Indiana were prepared and examined for their effects on various biological activities of VSV, including in vitro infection, hemagglutination, adsorption to cells, and mediation of cell fusion. Many enveloped viruses including vesicular stomatitis virus (VSV), family Rhabdoviridae, genus Vesiculovirus, transfer their nucleocapsids to the cytoplasm of host cells by the adsorption and receptor-mediated endocytosis, followed by fusion with the endosomal membrane [20, 21] . In the present study, we prepared thirteen MAbs specific for seven distinct epitopes on G protein of VSV-Indiana and examined for their effects on various biological activities of VSV including in vitro infection, HA, adsorption to the cells, and mediation of cell-cell fusion.

title:
  Identification of epitopes associated with different biological activities on the glycoprotein of vesicular stomatitis virus by use of monoclonal antibodies

abstract:
  Thirteen monoclonal antibodies (MAbs) to the glycoprotein (G) of vesicular stomatitis virus (VSV) serotype Indiana were prepared and examined for their effects on various biological activities of VSV, including in vitro infection, hemagglutination, adsorption to cells, and mediation of cell fusion. Competitive binding assays with these MAbs revealed the presence of at least seven distinct antigenic determinants (epitopes) on the G protein. In some cases, overlappings among epitopes to various degrees were observed as partial inhibition or binding enhancement. The MAbs to all the epitopes but one (epitopes 1–6) reacted with the denatured G protein in a Western immunoblot analysis. Four of the epitopes (epitopes 2, 4, 5, and 7) were involved in neutralization and two (epitopes 1 and 2) in hemagglutination inhibition. None of the MAbs inhibited the adsorption of radiolabeled VSV to BHK-21 cells; the MAbs to epitope 2 slightly enhanced the virus adsorption. All neutralization epitopes except epitope 2 (epitopes 4, 5, and 7) were associated with inhibition of VSV-mediated cell fusion. These results show a direct spatial relationship between the epitopes recognized by the MAbs and functional sites on G protein and further insights into the structure and function of G protein.


id: cord-008556-oetrdm8g
title: Regulation of Protein Synthesis in Virus-Infected Animal Cells
author: Kozak, Marilyn
date: 2008-03-01
txt: ./txt/cord-008556-oetrdm8g.txt
cache: ./cache/cord-008556-oetrdm8g.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131717/
doi: 10.1016/s0065-3527(08)60265-1
summary:
  One consequence of the scanning mechanism is that deleting the "ribosome binding site" (i.e., the normal initiator codon and flanking sequences) will not abolish translation; ribosomes will simply use the next AUG codon downstream, which, in some cases, has been shown to direct the synthesis of a biologically active, truncated protein (Downey et al., 1984; Halpern and Smiley, 1984; Katinka and Yaniv, 1982) . The best evidence for this is the ability of both EMC and SFV 26 S mRNA to be translated in EMC virus-infected cells, in which host translation is drastically inhibited by a mechanism that has not been difined, but that clearly does not involve cap binding protein (Mosenkis et al., 1985) . In wild-type adenovirus-infected cells, in which host protein synthesis is drastically reduced, both adenovirus and influenza virus mRNAs are translated efficiently.

abstract:
  This chapter summarizes the structural features that govern the translation of viral mRNAs: where the synthesis of a protein starts and ends, how many proteins can be produced from one mRNA, and how efficiently. It focuses on the interplay between viral and cellular mRNAs and the translational machinery. That interplay, together with the intrinsic structure of viral mRNAs, determines the patterns of translation in infected cells. It also points out some possibilities for translational regulation that can only be glimpsed at present, but are likely to come into focus in the future. The mechanism of selecting the initiation site for protein synthesis appears to follow a single formula. The translational machinery displays a certain flexibility that is exploited more frequently by viral than by cellular mRNAs. Although some of the parameters that determine efficiency have been identified, how efficiently a given mRNA will be translated cannot be predicted by summing the known parameters.


id: cord-011435-x73foqu7
author: Glanz, Anna
date: 2020-04-14
txt: ./txt/cord-011435-x73foqu7.txt
cache: ./cache/cord-011435-x73foqu7.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7232324/
doi: 10.3390/v12040442
summary:
  title: High Throughput Screening of FDA-Approved Drug Library Reveals the Compounds that Promote IRF3-Mediated Pro-Apoptotic Pathway Inhibit Virus Replication Previously, we uncovered a function for nontranscriptional IRF3 (nt-IRF3), RLR (RIG-I-like receptor)-induced IRF3-mediated pathway of apoptosis (RIPA), which triggers apoptotic killing of virus-infected cells. In contrast to the transcriptional pathway, nt-Irf3 in virus-infected cells functions as a chaperone protein by translocating the pro-apoptotic protein BCL2-associated X (BAX) to the mitochondria, thereby causing apoptotic cell death, which we named RLR (RIG-I-like receptor)-induced IRF3-mediated pathway of apoptosis (RIPA) ( Figure 1A ) [7] [8] [9] [10] [11] [12] [13] . We validated these results by immunoblot analyses, which demonstrate that doxorubicin treatment inhibited the expression of VSV-G protein, a viral envelope glycoprotein as well as the virus-encoded GFP ( Figure 3B ). We validated these results by immunoblot analyses, which demonstrate that doxorubicin treatment inhibited the expression of VSV-G protein, a viral envelope glycoprotein as well as the virus-encoded GFP ( Figure 3B ).

title:
  High Throughput Screening of FDA-Approved Drug Library Reveals the Compounds that Promote IRF3-Mediated Pro-Apoptotic Pathway Inhibit Virus Replication

abstract:
  Interferon (IFN) regulatory factor 3 (IRF3) is the key transcription factor for the induction of IFN and antiviral genes. The absence of antiviral genes in IRF3 deficiency leads to susceptibility to a wide range of viral infections. Previously, we uncovered a function for nontranscriptional IRF3 (nt-IRF3), RLR (RIG-I-like receptor)-induced IRF3-mediated pathway of apoptosis (RIPA), which triggers apoptotic killing of virus-infected cells. Using knock-in mice expressing a transcriptionally inactive, but RIPA-active, IRF3 mutant, we demonstrated the relative contribution of RIPA to host antiviral defense. Given that RIPA is a cellular antiviral pathway, we hypothesized that small molecules that promote RIPA in virus-infected cells would act as antiviral agents. To test this, we conducted a high throughput screen of a library of FDA-approved drugs to identify novel RIPA activators. Our screen identified doxorubicin as a potent RIPA-activating agent. In support of our hypothesis, doxorubicin inhibited the replication of vesicular stomatitis virus, a model rhabdovirus, and its antiviral activity depended on its ability to activate IRF3 in RIPA. Surprisingly, doxorubicin inhibited the transcriptional activity of IRF3. The antiviral activity of doxorubicin was expanded to flavivirus and herpesvirus that also activate IRF3. Mechanistically, doxorubicin promoted RIPA by activating the extracellular signal-regulated kinase (ERK) signaling pathway. Finally, we validated these results using another RIPA-activating compound, pyrvinium pamoate, which showed a similar antiviral effect without affecting the transcriptional activity of IRF3. Therefore, we demonstrate that the RIPA branch of IRF3 can be targeted therapeutically to prevent virus infection.


id: cord-020714-h1fevqcw
title: Membrane Glycoproteins of Enveloped Viruses
author: Compans, Richard W.
date: 2008-05-30
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7146817/
doi: 10.1016/s0070-2161(08)60750-9
abstract:
  This chapter focuses on the recent information of the glycoprotein components of enveloped viruses and points out specific findings on viral envelopes. Although enveloped viruses of different major groups vary in size and shape, as well as in the molecular weight of their structural polypeptides, there are general similarities in the types of polypeptide components present in virions. The types of structural components found in viral membranes are summarized briefly in the chapter. All the enveloped viruses studied to date possess one or more glycoprotein species and lipid as a major structural component. The presence of carbohydrate covalently linked to proteins is demonstrated by the incorporation of a radioactive precursor, such as glucosamine or fucose, into viral polypeptides, which is resolved by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Enveloped viruses share many common features in the organization of their structural components, as indicated by several approaches, including electron microscopy, surface-labeling, and proteolytic digestion experiments, and the isolation of subviral components. The chapter summarizes the detailed structure of the glycoproteins of four virus groups: (1) influenza virus glycoproteins, (2) rhabdovirus G protein, (3) togavirus glycoprotein, and (4) paramyxovirus glycoproteins The information obtained includes the size and shape of viral glycoproteins, the number of polypeptide chains in the complete glycoprotein structure, and compositional data on the polypeptide and oligosaccharide portions of the molecules.


id: cord-104239-xxlcdbqi
title: The organization of endoplasmic reticulum export complexes
author: nan
date: 1996-10-01
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2121027/
doi: nan
abstract:
  Export of cargo from the ER occurs through the formation of 60-70nm COPII-coated vesicular carriers. We have applied serial-thin sectioning and stereology to quantitatively characterize the three-dimensional organization of ER export sites in vivo and in vitro. We find that ER buds in vivo are nonrandomly distributed, being concentrated in regional foci we refer to as export complexes. The basic organization of an export complex can be divided into an active COPII-containing budding zone on a single ER cisterna, which is adjacent to budding zones found on distantly connected ER cisternae. These budding foci surround and face a central cluster of morphologically independent vesicular-tubular elements that contain COPI coats involved in retrograde transport. Vesicles within these export complexes contain concentrated cargo molecules. The structure of vesicular-tubular clusters in export complexes is particularly striking in replicas generated using a quick-freeze, deep-etch approach to visualize for the first time their three-dimensional organization and cargo composition. We conclude that budding from the ER through recruitment of COPII is confined to highly specialized export complexes that topologically restrict anterograde transport to regional foci to facilitate efficient coupling to retrograde recycling by COPI.


id: cord-262752-bwofzbwa
title: Current status on the development of pseudoviruses for enveloped viruses
author: Li, Qianqian
date: 2017-12-07
txt: ./txt/cord-262752-bwofzbwa.txt
cache: ./cache/cord-262752-bwofzbwa.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1002/rmv.1963
doi: 10.1002/rmv.1963
summary:
  Early work by Witte and colleagues showed that when they used VSV to infect the cells in which MLV is packaged, they were able to harvest pseudovirus for use in neutralization antibody assays. Development of in vitro and in vivo rabies virus neutralization assays based on a high-titer pseudovirus system Development of a pseudotyped-lentiviral-vector-based neutralization assay for chikungunya virus infection Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase Use of vesicular stomatitis virus pseudotypes bearing Hantaan or Seoul virus envelope proteins in a rapid and safe neutralization test A neutralization test for specific detection of Nipah virus antibodies using pseudotyped vesicular stomatitis virus expressing green fluorescent protein Truncation of the human immunodeficiency virus-type-2 envelope glycoprotein allows efficient pseudotyping of murine leukemia virus retroviral vector particles Cholesterol supplementation during production increases the infectivity of retroviral and Lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G)

abstract:
  Emerging and reemerging infectious diseases have a strong negative impact on public health. However, because many of these pathogens must be handled in biosafety level, 3 or 4 containment laboratories, research and development of antivirals or vaccines against these diseases are often impeded. Alternative approaches to address this issue have been vigorously pursued, particularly the use of pseudoviruses in place of wild‐type viruses. As pseudoviruses have been deprived of certain gene sequences of the virulent virus, they can be handled in biosafety level 2 laboratories. Importantly, the envelopes of these viral particles may have similar conformational structures to those of the wild‐type viruses, making it feasible to conduct mechanistic investigation on viral entry and to evaluate potential neutralizing antibodies. However, a variety of challenging issues remain, including the production of a sufficient pseudovirus yield and the inability to produce an appropriate pseudotype of certain viruses. This review discusses current progress in the development of pseudoviruses and dissects the factors that contribute to low viral yields.


id: cord-262753-jld1ygxt
title: Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
author: Neidermyer, William J.
date: 2019-06-21
url: https://www.ncbi.nlm.nih.gov/pubmed/31226162/
doi: 10.1371/journal.ppat.1007875
abstract:
  Infection of mammalian cells with vesicular stomatitis virus (VSV) results in the inhibition of cellular translation while viral translation proceeds efficiently. VSV RNA synthesis occurs entirely within the cytoplasm, where during transcription the viral polymerase produces 5 mRNAs that are structurally indistinct to cellular mRNAs with respect to their 5′ cap-structure and 3′-polyadenylate tail. Using the global approach of massively parallel sequencing of total cytoplasmic, monosome- and polysome-associated mRNA, we interrogate the impact of VSV infection of HeLa cells on translation. Analysis of sequence reads in the different fractions shows >60% of total cytoplasmic and polysome-associated reads map to the 5 viral genes by 6 hours post-infection, a time point at which robust host cell translational shut-off is observed. Consistent with an overwhelming abundance of viral mRNA in the polysome fraction, the reads mapping to cellular genes were reduced. The cellular mRNAs that remain most polysome-associated following infection had longer half-lives, were typically larger, and were more AU rich, features that are shared with the viral mRNAs. Several of those mRNAs encode proteins known to positively affect viral replication, and using chemical inhibition and siRNA depletion we confirm that the host chaperone heat shock protein 90 (hsp90) and eukaryotic translation initiation factor 3A (eIF3A)—encoded by 2 such mRNAs—support viral replication. Correspondingly, regulated in development and DNA damage 1 (Redd1) encoded by a host mRNA with reduced polysome association inhibits viral infection. These data underscore the importance of viral mRNA abundance in the shut-off of host translation in VSV infected cells and link the differential translatability of some cellular mRNAs with pro- or antiviral function.


id: cord-267712-mhx8e5y0
title: Evaluation of attenuated VSVs with mutated M or/and G proteins as vaccine vectors
author: Fang, Xinkui
date: 2012-02-08
txt: ./txt/cord-267712-mhx8e5y0.txt
cache: ./cache/cord-267712-mhx8e5y0.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/22222871/
doi: 10.1016/j.vaccine.2011.12.085
summary:
  Due to its potent capabilities in triggering cellular, humoral, and mucosal immunities in animals, even after a single administration, recombinant VSV has been studied as a vaccine vector not only for preventing vesicular stomatitis disease in livestock [4] , but a number of human pathogens including: Influenza virus, Ebola virus, Marburg virus, Human immunodeficiency (HIV) virus, Severe Acute Respiratory Syndrome (SARS) virus, and Hepatitis C virus [5] [6] [7] [8] [9] . In vivo, however, VSV M protein mutant proved to be only moderately attenuated in experimental infections [16, 21] , whereas there is currently no information available if recombinant VSV with truncated G protein is safe or not when animals challenged with high dose of the mutant virus. Based on pathogenicity and capabilities to stimulate potent immune responses, we aimed to identify a suitable recombinant VSV vaccine vector and vaccine candidate for preventing vesicular stomatitis disease.

abstract:
  Vesicular stomatitis virus (VSV) is a promising vector for vaccine and oncolysis, but it can also produce acute diseases in cattle, horses, and swine characterized by vesiculation and ulceration of the tongue, oral tissues, feet, and teats. In experimental animals (primates, rats, and mice), VSV has been shown to lead to neurotoxicities, such as hind limb paralysis. The virus matrix protein (M) and glycoprotein (G) are both major pathogenic determinants of wild-type VSV and have been the major targets for the production of attenuated strains. Existing strategies for attenuation included: (1) deletion or M51R substitution in the M protein (VSVΔM51 or VSVM51R, respectively); (2) truncation of the C-terminus of the G protein (GΔ28). Despite these mutations, recombinant VSV with mutated M protein is only moderately attenuated in animals, whereas there are no detailed reports to determine the pathogenicity of recombinant VSV with truncated G protein at high dose. Thus, a novel recombinant VSV (VSVΔM51-GΔ28) as well as other attenuated VSVs (VSVΔM51, VSV-GΔ28) were produced to determine their efficacy as vaccine vectors with low pathogenicity. In vitro studies indicated that truncated G protein (GΔ28) could play a more important role than deletion of M51 (ΔM51) for attenuation of recombinant VSV. VSVΔM51-GΔ28 was determined to be the most attenuated virus with low pathogenicity in mice, with VSV-GΔ28 also showing relatively reduced pathogenicity. Further, neutralizing antibodies stimulated by VSV-GΔ28 proved to be significantly higher than in mice treated with VSVΔM51-GΔ28. In conclusion, among different attenuated VSVs with mutated M and/or G proteins, recombinant VSV with only truncated G protein (VSV-GΔ28) demonstrated ideal balance between pathogenesis and stimulating a protective immune response. These properties make VSV-GΔ28 a promising vaccine vector and vaccine candidate for preventing vesicular stomatitis disease.


id: cord-268565-2sg1tlrg
title: Recombinant vesicular stomatitis virus as an HIV-1 vaccine vector
author: Clarke, David K.
date: 2006-09-15
txt: ./txt/cord-268565-2sg1tlrg.txt
cache: ./cache/cord-268565-2sg1tlrg.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/16977404/
doi: 10.1007/s00281-006-0042-3
summary:
  However, because durable neutralizing antibodies are usually elicited against the VSV surface glycoprotein after a single vaccination with replication-competent rVSV vectors, glycoprotein exchange vectors were designed to allow more effective boosting of immune responses to target antigens. In these pioneering studies, rhesus macaques were immunized with a combination of two prototypic rVSV vectors expressing a HIV-1 89.6 Env gp160/VSV-G fusion polypeptide and simian immunodeficiency virus (SIV) Gag p55 protein. Vaccine vector administration by a combination of intramuscular and intranasal routes clearly demonstrated the ability of rVSV vectors to elicit potent antigen-specific cellmediated immune responses in NHPs. In addition, this study also clearly demonstrated for the first time the ability of rVSV vectors expressing Env and Gag proteins to provide highly significant protection from AIDS after an intravenous heterologous SIV/HIV (SHIV) 89 .6P challenge [89] . Immunogenicity of attenuated vesicular stomatitis virus vectors expressing HIV type 1 Env and SIV Gag proteins: comparison of intranasal and intramuscular vaccination routes

abstract:
  Recombinant vesicular stomatitis virus (rVSV) is currently under evaluation as a human immunodeficiency virus (HIV)-1 vaccine vector. The most compelling reasons to develop rVSV as a vaccine vector include a very low seroprevalence in humans, the ability to infect and robustly express foreign antigens in a broad range of cells, and vigorous growth in continuous cell lines used for vaccine manufacture. Numerous preclinical studies with rVSV vectors expressing antigens from a variety of human pathogens have demonstrated the versatility, flexibility, and potential efficacy of the rVSV vaccine platform. When administered to nonhuman primates (NHPs), rVSV vectors expressing HIV-1 Gag and Env elicited robust HIV-1-specific cellular and humoral immune responses, and animals immunized with rVSV vectors expressing simian immunodeficiency virus (SIV) Gag and HIV Env were protected from AIDS after challenge with a pathogenic SIV/HIV recombinant. However, results from an exploratory neurovirulence study in NHPs indicated that these prototypic rVSV vectors might not be adequately attenuated for widespread use in human populations. To address this safety concern, a variety of different attenuation strategies, designed to produce a range of further attenuated rVSV vectors, are currently under investigation. Additional modifications of further attenuated rVSV vectors to upregulate expression of HIV-1 antigens and coexpress molecular adjuvants are also being developed in an effort to balance immunogenicity and attenuation.


id: cord-270380-1me7ugkg
author: Wang, Xiaona
date: 2020-03-19
txt: ./txt/cord-270380-1me7ugkg.txt
cache: ./cache/cord-270380-1me7ugkg.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.3390/v12030335
doi: 10.3390/v12030335
summary:
  Both proteins exhibited effective anti-vesicular stomatitis virus (VSV) activity in Vero, F81 (feline kidney cell), Madin–Darby bovine kidney (MDBK), Madin–Darby canine kidney (MDCK), and porcine kidney (PK-15) cells, showing broader cross-species antiviral activity than the INTERCAT IFN antiviral drug. Furthermore, the recombinant feIFN-ωa and feIFN-ωb proteins demonstrated antiviral activity against VSV, feline coronavirus (FCoV), canine parvovirus (CPV), bovine viral diarrhea virus (BVDV), and porcine epidemic diarrhea virus (PEDV), indicating better broad-spectrum antiviral activity than the INTERCAT IFN. As shown in Figure 6 , the purified recombinant feIFN-ωa and feIFN-ωb proteins showed antiviral activity in both homologous animal cells (F81 cells) and heterologous animal cells (Vero, MDCK, MDBK, and PK-15 cells) in vitro. Our data showed that purified recombinant feIFN-ωa and feIFN-ωb had broad-spectrum antiviral activity in homologous and heterologous animal cells, suggesting they are candidates for the development of effective therapeutic agents to be used against viral infections in pet cats.

title:
  Cloning, Prokaryotic Soluble Expression, and Analysis of Antiviral Activity of Two Novel Feline IFN-ω Proteins

abstract:
  Cats are becoming more popular as household companions and pets, forming close relationships with humans. Although feline viral diseases can pose serious health hazards to pet cats, commercialized preventative vaccines are lacking. Interferons (IFNs), especially type I IFNs (IFN-α, IFN-β, and interferon omega (IFN-ω)), have been explored as effective therapeutic drugs against viral diseases in cats. Nevertheless, there is limited knowledge regarding feline IFN-ω (feIFN-ω), compared to IFN-α and IFN-β. In this study, we cloned the genes encoding feIFN-ωa and feIFN-ωb from cat spleen lymphocytes. Homology and phylogenetic tree analysis revealed that these two genes belonged to new subtypes of feIFN-ω. The recombinant feIFN-ωa and feIFN-ωb proteins were expressed in their soluble forms in Escherichia coli, followed by purification. Both proteins exhibited effective anti-vesicular stomatitis virus (VSV) activity in Vero, F81 (feline kidney cell), Madin–Darby bovine kidney (MDBK), Madin–Darby canine kidney (MDCK), and porcine kidney (PK-15) cells, showing broader cross-species antiviral activity than the INTERCAT IFN antiviral drug. Furthermore, the recombinant feIFN-ωa and feIFN-ωb proteins demonstrated antiviral activity against VSV, feline coronavirus (FCoV), canine parvovirus (CPV), bovine viral diarrhea virus (BVDV), and porcine epidemic diarrhea virus (PEDV), indicating better broad-spectrum antiviral activity than the INTERCAT IFN. The two novel feIFN-ω proteins (feIFN-ωa and feIFN-ωb) described in this study show promising potential to serve as effective therapeutic agents for treating viral infections in pet cats.


id: cord-272051-arz8r204
author: Federico, Maurizio
date: 2011-08-15
txt: ./txt/cord-272051-arz8r204.txt
cache: ./cache/cord-272051-arz8r204.txt
extension: .txt
mime: text/plain
url: https://www.sciencedirect.com/science/article/pii/S0042682211002157
doi: 10.1016/j.virol.2011.05.002
summary:
  Since the replication of many virus species requires the activity of host cell proteases, investigating the effects of PIs on the life cycle of viruses other than HIV would be of interest. Considering that PIs are well tolerated drugs in vivo, and that many relevant human pathogens belong to the family of RNA viruses infecting cells through an endocytic pathway, this finding would open the way towards a broader therapeutic use of PIs. HIV virions emerging from cells treated with PIs remain immature viral particles as a consequence of the block of Gag polyprotein cleavage. Cells were treated overnight with the PI doses most effective against VSV and/or influenza virus replication, then labeled with LysoSensor Green DND-189 for 30 min in the presence of PIs, and finally analyzed by FACS.

title:
  HIV-protease inhibitors block the replication of both vesicular stomatitis and influenza viruses at an early post-entry replication step

abstract:
  The inhibitors of HIV-1 protease (PIs) have been designed to block the activity of the viral aspartyl-protease. However, it is now accepted that this family of inhibitors can also affect the activity of cell proteases. Since the replication of many virus species requires the activity of host cell proteases, investigating the effects of PIs on the life cycle of viruses other than HIV would be of interest. Here, the potent inhibition induced by saquinavir and nelfinavir on the replication of both vesicular stomatitis and influenza viruses is described. These are unrelated enveloped RNA viruses infecting target cells upon endocytosis and intracellular fusion. The PI-induced inhibition was apparently a consequence of a block at the level of the fusion between viral envelope and endosomal membranes. These findings would open the way towards the therapeutic use of PIs against enveloped RNA viruses other than HIV.


id: cord-275348-jna496x7
author: Kapadia, Sagar U.
date: 2008-06-20
txt: ./txt/cord-275348-jna496x7.txt
cache: ./cache/cord-275348-jna496x7.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/S0042682208001736
doi: 10.1016/j.virol.2008.03.002
summary:
  A SARS vaccine based on a live-attenuated vesicular stomatitis virus (VSV) recombinant expressing the SARS-CoV S protein provides long-term protection of immunized mice from SARS-CoV infection (Kapadia, S.U., Rose, J. We found that the vaccine given intramuscularly induced a neutralizing antibody response to SARS-CoV that was approximately ten-fold greater than that required for the protection from SARS-CoV infection, and significantly greater than that generated by the replication-competent vector expressing SARS-CoV S protein given by the same route. In order to evaluate this vector as a SARS vaccine candidate, we also developed a SARS-CoV neutralization assay using a pseudotyped VSV recombinant expressing a green fluorescent protein. SARS-CoV neutralizing antibody titers of these sera were determined by incubating VSVΔG-EGFP/SΔtail-HA virus with serial dilutions of these sera, and the virusserum mixtures were transferred to a monolayer of Vero E6 cells.

title:
  SARS vaccine based on a replication-defective recombinant vesicular stomatitis virus is more potent than one based on a replication-competent vector

abstract:
  A SARS vaccine based on a live-attenuated vesicular stomatitis virus (VSV) recombinant expressing the SARS-CoV S protein provides long-term protection of immunized mice from SARS-CoV infection (Kapadia, S.U., Rose, J. K., Lamirande, E., Vogel, L., Subbarao, K., Roberts, A., 2005. Long-term protection from SARS coronavirus infection conferred by a single immunization with an attenuated VSV-based vaccine. Virology 340(2), 174–82.). Because it is difficult to obtain regulatory approval of vaccine based on live viruses, we constructed a replication-defective single-cycle VSV vector in which we replaced the VSV glycoprotein (G) gene with the SARS-CoV S gene. The virus was only able to infect cells when pseudotyped with the VSV G protein. We measured the effectiveness of immunization with the single-cycle vaccine in mice. We found that the vaccine given intramuscularly induced a neutralizing antibody response to SARS-CoV that was approximately ten-fold greater than that required for the protection from SARS-CoV infection, and significantly greater than that generated by the replication-competent vector expressing SARS-CoV S protein given by the same route. Our results, along with earlier studies showing potent induction of T-cell responses by single-cycle vectors, indicate that these vectors are excellent alternatives to live-attenuated VSV.


id: cord-276009-p98wjtjb
author: Iyer, Arun V.
date: 2009-02-05
url: https://www.ncbi.nlm.nih.gov/pubmed/19070640/
doi: 10.1016/j.vaccine.2008.11.087
title:
  Recombinant vesicular stomatitis virus-based west Nile vaccine elicits strong humoral and cellular immune responses and protects mice against lethal challenge with the virulent west Nile virus strain LSU-AR01

abstract:
  Vesicular stomatitis virus (VSV) has been extensively utilized as a viral vector system for the induction of protective immune responses against a variety of pathogens. We constructed recombinant VSVs specifying either the Indiana or Chandipura virus G glycoprotein and expressing the West Nile virus (WNV) envelope (E) glycoprotein. Mice were intranasally vaccinated using a prime (Indiana)-boost (Chandipura) immunization approach and challenged with the virulent WNV-LSU-AR01. Ninety-percent (9 of 10) of the vaccinated mice survived as compared to 10% of the mock-vaccinated mice after WNV lethal challenge. Histopathological examination of brain tissues revealed neuronal necrosis in mock-vaccinated mice but not in vaccinated mice, and vaccinated, but not mock-vaccinated mice developed a strong neutralizing antibody response against WNV. Extensive immunological analysis using polychromatic flow cytometry staining revealed that vaccinated, but not mock-vaccinated mice developed robust cellular immune responses as evidenced by up-regulation of CD4(+) CD154(+) IFNγ(+) T cells in vaccinated, but not mock-vaccinated mice. Similarly, vaccinated mice developed robust E-glycoprotein-specific CD8(+) T cell immune responses as evidenced by the presence of a high percentage of CD8(+) CD62L(low) IFNγ(+) cells. In addition, a sizeable population of CD8(+) CD69(+) cells was detected indicating E-specific activation of mature T cells and CD4(+) CD25(+) CD127(low) T regulatory (T reg) cells were down-regulated. These results suggest that VSV-vectored vaccines administered intranasally can efficiently induce protective humoral and cellular immune responses against WNV infections.


id: cord-277823-vijh6x1l
author: TERAMICHI, Takurou
date: 2019-11-05
txt: ./txt/cord-277823-vijh6x1l.txt
cache: ./cache/cord-277823-vijh6x1l.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1292/jvms.19-0436
doi: 10.1292/jvms.19-0436
summary:
  A serological survey of Middle East respiratory syndrome coronavirus (MERS-CoV) was conducted among dromedary camels and herbivorous animals sharing the same pasturage in Ethiopia. One of camel serum that showed a high antibody titer in the neutralization test by live MERS-CoV was treated as a positive control. According to the results of the previous study, antibody titers of ≥16 are treated as positive in neutralization test using VSV-MERS/GFP. Cows that were antibody positive in the neutralization test using VSV-MERS/GFP or cELISA were different animals and both were antibody negative in the neutralization test using MERS-CoV. S1-ELISA was not sensitive compared to other tests because only 16 serum samples were positive and they required an antibody titer of ≥64 in VSV-MERS/GFP. The present study shows that the neutralization test using VSV-MERS/GFP, S1-ELISA, and cELISA are as specific to MERS-CoV infection as the serological tests, although their sensitivities slightly differ. Middle East respiratory syndrome coronavirus neutralising serum antibodies in dromedary camels: a comparative serological study

title:
  Evaluation of serological assays available in a biosafety level 2 laboratory and their application for survey of Middle East respiratory syndrome coronavirus among livestock in Ethiopia

abstract:
  A serological survey of Middle East respiratory syndrome coronavirus (MERS-CoV) was conducted among dromedary camels and herbivorous animals sharing the same pasturage in Ethiopia. The pseudotyped vesicular stomatitis virus coated with the spike protein of MERS-CoV was used in virus neutralization (VN) tests performed in a biosafety level (BSL)-2 laboratory. The results were similar to those obtained from the VN test using live MERS-CoV and were more sensitive than the ELISA performed using synthetic MERS S1 fragment as the antigen as well as the competitive ELISA performed using a monoclonal antibody against MERS-CoV. According to the comprehensive results of the four types of serodiagnosis methods, positive antibodies were detected only in dromedary camels and the remaining herbivorous animals were not infected with the virus. Moreover, using the present procedure, serological tests for MERS-CoV can be conducted even in BSL 2 laboratory.


id: cord-285749-0ejhd9nw
author: Hoffmann, Markus
date: 2016-03-30
txt: ./txt/cord-285749-0ejhd9nw.txt
cache: ./cache/cord-285749-0ejhd9nw.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1371/journal.pone.0152134
doi: 10.1371/journal.pone.0152134
summary:
  Generation of VSV pseudotypes (VSVpp) was performed as follows: HEK-293T cells were transfected by calcium-phosphate precipitation with expression plasmids encoding viral surface proteins, VSV-G (positive control) , NiV-F/G, FLUAV-HA and/or NA and bat-FLUAV-HAL and/or NAL, or empty plasmid (pCAGGS) as negative control. In order to investigate the potential of human TTSPs to proteolytically activate batFLUAV-HAL for host cell entry, we additionally cotransfected the cells with expression plasmids for TMPRSS2, DESC-1 or MSPL. Notably, three bat cell lines (EidNi/41, HypNi/1.1 and EpoNi/22.1) were susceptible to entry of pseudotypes bearing HAL and NAL of batFLUAV (Fig 2B) , demonstrating that surface glycoproteins of batFLUAV can mediate cellular entry. To assess proteolytic activation of HA/HAL proteins, vesicular stomatitis virus-based pseudotypes (VSVpp) were produced in cells transfected to express the indicated type II transmembrane serine proteases (B) or different amounts of TMPRSS2 (C).

title:
  The Hemagglutinin of Bat-Associated Influenza Viruses Is Activated by TMPRSS2 for pH-Dependent Entry into Bat but Not Human Cells

abstract:
  New World bats have recently been discovered to harbor influenza A virus (FLUAV)-related viruses, termed bat-associated influenza A-like viruses (batFLUAV). The internal proteins of batFLUAV are functional in mammalian cells. In contrast, no biological functionality could be demonstrated for the surface proteins, hemagglutinin (HA)-like (HAL) and neuraminidase (NA)-like (NAL), and these proteins need to be replaced by their human counterparts to allow spread of batFLUAV in human cells. Here, we employed rhabdoviral vectors to study the role of HAL and NAL in viral entry. Vectors pseudotyped with batFLUAV-HAL and -NAL were able to enter bat cells but not cells from other mammalian species. Host cell entry was mediated by HAL and was dependent on prior proteolytic activation of HAL and endosomal low pH. In contrast, sialic acids were dispensable for HAL-driven entry. Finally, the type II transmembrane serine protease TMPRSS2 was able to activate HAL for cell entry indicating that batFLUAV can utilize human proteases for HAL activation. Collectively, these results identify viral and cellular factors governing host cell entry driven by batFLUAV surface proteins. They suggest that the absence of a functional receptor precludes entry of batFLUAV into human cells while other prerequisites for entry, HAL activation and protonation, are met in target cells of human origin.


id: cord-286390-ytgw3j4s
author: Case, James Brett
date: 2020-07-03
txt: ./txt/cord-286390-ytgw3j4s.txt
cache: ./cache/cord-286390-ytgw3j4s.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/S1931312820303620
doi: 10.1016/j.chom.2020.06.021
summary:
  An anticipated correlate of such countermeasures is the level of neutralizing antibodies against the SARS-CoV-2 spike protein, which engages with host ACE2 receptor for entry. Using an infectious molecular clone of vesicular stomatitis virus (VSV) expressing eGFP as a marker of infection, we replaced the glycoprotein gene (G) with the spike protein of SARS-CoV-2 (VSV-eGFP-SARS-CoV-2) and developed a high-throughput imaging-based neutralization assay at biosafety level 2. We engineered an infectious molecular clone of vesicular 83 stomatitis virus (VSV) to encode the SARS-CoV-2 S protein in place of the native envelope 84 glycoprotein (G) and rescued an autonomously replication-competent virus bearing the spike. Evaluation of a novel vesicular stomatitis virus pseudotype-based assay for detection of 641 neutralizing antibody responses to SARS-CoV Vesicular stomatitis virus pseudotyped with severe acute 645 respiratory syndrome coronavirus spike protein Retroviruses pseudotyped with the severe acute respiratory 722 syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting 723 enzyme 2

title:
  Neutralizing antibody and soluble ACE2 inhibition of a replication-competent VSV-SARS-CoV-2 and a clinical isolate of SARS-CoV-2.

abstract:
  Abstract Antibody-based interventions against SARS-CoV-2 could limit morbidity, mortality, and possibly transmission. An anticipated correlate of such countermeasures is the level of neutralizing antibodies against the SARS-CoV-2 spike protein, which engages with host ACE2 receptor for entry. Using an infectious molecular clone of vesicular stomatitis virus (VSV) expressing eGFP as a marker of infection, we replaced the glycoprotein gene (G) with the spike protein of SARS-CoV-2 (VSV-eGFP-SARS-CoV-2) and developed a high-throughput imaging-based neutralization assay at biosafety level 2. We also developed a focus-reduction neutralization test with a clinical isolate of SARS-CoV-2 at biosafety level 3. Comparing the neutralizing activities of various antibodies and ACE2-Fc soluble decoy protein in both assays revealed a high degree of concordance. These assays will help define correlates of protection for antibody-based countermeasures and vaccines against SARS-CoV-2. Additionally, replication-competent VSV-eGFP-SARS-CoV-2 provides a tool for testing inhibitors of SARS-CoV-2 mediated entry under reduced biosafety containment.


id: cord-290243-m8yfugr0
author: Kim, Kyung Ran
date: 2007-01-01
txt: ./txt/cord-290243-m8yfugr0.txt
cache: ./cache/cord-290243-m8yfugr0.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/S096808960600811X
doi: 10.1016/j.bmc.2006.09.066
summary:
  The resulting residue was purified by silica gel column chromatography using hexane and ethyl acetate (5:1) as the eluent to give the corresponding alcohol 8 (5.570 g, 95%) as a colorless oil: ½a To a stirred solution of alcohol 8 (3.050 g, 6.19 mmol) in anhydrous CH 2 Cl 2 (20 mL) was dropwise added (dieth-ylamino)sulfur trifluoride (DAST, 1.23 mL, 9.31 mmol) at À10°C and the reaction mixture was stirred at the same temperature for 30 min. After the volatiles were removed in vacuo, the resulting residue was purified by silica gel column chromatography using hexane and ethyl acetate (3:1) as the eluent to give To a stirred solution of 13 (110 mg, 0.19 mmol) in methanol (4.5 mL) and CH 2 Cl 2 (1.5 mL) was added 1 M NaOMe (0.40 mL, 0.40 mmol, in MeOH) at 0°C and the reaction mixture was stirred for 6 h at room temperature.

title:
  Design, synthesis, and biological evaluation of novel iso-d-2′,3′-dideoxy-3′-fluorothianucleoside derivatives

abstract:
  Abstract Novel iso-d-2′,3′-dideoxythianucleoside derivatives 1–4 were designed and asymmetrically synthesized as a bioisostere of lamivudine to search for new anti-HIV agents. The information about using sulfur participation occurred on DAST fluorination and Mitsunobu reaction will be of great help in synthesizing sulfur-containing compounds. Final compounds 1–4 were evaluated against HIV-1 and 2, HSV-1 and 2, EMCV, Cox. B3, VSV, FluA (Taiwan), FluA (Johan.), FCV, and FIP. Only cytosine analogue 3 showed a potent anti-VSV activity (EC50 =9.43μg/mL). This result implies that iso-2′,3′-dideoxy sugar templates might play a role of a sugar surrogate of nucleosides for the development of anti-RNA virus agent.


id: cord-291323-kbjyd5g3
title: Inhibition of PIKfyve kinase prevents infection by Zaire ebolavirus and SARS-CoV-2
author: Kang, Yuan-Lin
date: 2020-08-25
txt: ./txt/cord-291323-kbjyd5g3.txt
cache: ./cache/cord-291323-kbjyd5g3.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/32764148/
doi: 10.1073/pnas.2007837117
summary:
  We describe here potent inhibitory effects on content release and infection by chimeric vesicular stomatitis virus (VSV) containing the envelope proteins of Zaire ebolavirus (VSV-ZEBOV) or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (VSV-SARS-CoV-2) elicited by Apilimod and Vacuolin-1, small-molecule inhibitors of the main endosomal phosphatidylinositol-3-phosphate/phosphatidylinositol 5-kinase, PIKfyve. We describe here potent inhibitory effects on content release and infection by chimeric vesicular stomatitis virus (VSV) containing the envelope proteins of Zaire ebolavirus (VSV-ZEBOV) or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (VSV-SARS-CoV-2) elicited by Apilimod and Vacuolin-1, small-molecule inhibitors of the main endosomal phosphatidylinositol-3-phosphate/phosphatidylinositol 5-kinase, PIKfyve. We have constructed chimeric forms of vesicular stomatitis virus (VSV) bearing the fusion proteins of Zaire ebolavirus (ZEBOV) or SARS coronavirus 2 (SARS-CoV-2) and shown that two small-molecule inhibitors of an endosomal lipid kinase (PIKfyve) inhibit viral infection by preventing release of the viral contents from endosomes.

abstract:
  Virus entry is a multistep process. It initiates when the virus attaches to the host cell and ends when the viral contents reach the cytosol. Genetically unrelated viruses can subvert analogous subcellular mechanisms and use similar trafficking pathways for successful entry. Antiviral strategies targeting early steps of infection are therefore appealing, particularly when the probability for successful interference through a common step is highest. We describe here potent inhibitory effects on content release and infection by chimeric vesicular stomatitis virus (VSV) containing the envelope proteins of Zaire ebolavirus (VSV-ZEBOV) or severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (VSV-SARS-CoV-2) elicited by Apilimod and Vacuolin-1, small-molecule inhibitors of the main endosomal phosphatidylinositol-3-phosphate/phosphatidylinositol 5-kinase, PIKfyve. We also describe potent inhibition of SARS-CoV-2 strain 2019-nCoV/USA-WA1/2020 by Apilimod. These results define tools for studying the intracellular trafficking of pathogens elicited by inhibition of PIKfyve kinase and suggest the potential for targeting this kinase in developing small-molecule antivirals against SARS-CoV-2.


id: cord-292593-apdyaujt
author: Coulter-Mackie, Marion
date: 1985-10-31
txt: ./txt/cord-292593-apdyaujt.txt
cache: ./cache/cord-292593-apdyaujt.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/0168170285900498
doi: 10.1016/0168-1702(85)90049-8
summary:
  Interference with vesicular stomatitis virus (VSV) production by JHMV was tested in persistently or latently infected RN-2 cells using as the controls, uninfected RN2-2 cells. Since upon infection of RN2-2 cultures with JHMV at 39.5"C and maintenance in a state of latency at that temperature for several days, the cells commenced yielding pfu upon shift down to 32.5"C (Lucas et al., 1978) , these data suggest that penetration and eclipse of the virus inoculum must have occurred normally at 39.5"C and restriction most probably developed at a subsequent stage. It is very significant that during incubation at the restrictive temperature for as long as 7 days and beyond, when RN2-2 cells had multiplied through 6-7 generations, the 56K antigen was readily detectable (channel I), implying that during latency, in the absence of infectious virus production, translation into JHMV nucleocapsid protein continued.

title:
  In vivo and in vitro models of demyelinating diseases XII. Persistence and expression of corona JHM vims functions in RN2-2 Schwannoma cells during latency

abstract:
  Abstract The coronavirus JHMV persistently infects rat Schwannoma cells RN2-2 at 32.5°C and enters a host-imposed reversible, latent state at 39.5°C. JHMV can remain up to 20 days in the latent state and about 14 days before the cultures lose the capacity to resume virus production upon return to 32.5°C. Although persistently and latently infected RN2-2 cells display resistance to superinfection by a heterologous agent VSV, these cells do not release detectable soluble mediators (e.g., interferon) of the antiviral state. Nevertheless, RN2-2 cells are competent to synthesize and release interferon when treated with the appropriate inducers. These observations suggest that interferon does not play any role or may not be the major factor in the control of latency in the Schwannoma cell. Hybridization with virus-specific cDNAs shows that all viral mRNAs are present during latency and that viral mRNAs are present in the polysomes of infected cells at 39.5°C. Western immunoblotting with hybridoma antibodies demonstrates that viral specific proteins are produced at the restrictive temperature. These results


id: cord-296187-nnv2e7gr
title: Bcr-Abl tyrosine kinase inhibitor imatinib as a potential drug for COVID-19
author: Mulgaonkar, Nirmitee
date: 2020-08-18
txt: ./txt/cord-296187-nnv2e7gr.txt
cache: ./cache/cord-296187-nnv2e7gr.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.06.18.158196
doi: 10.1101/2020.06.18.158196
summary:
  The SARS-CoV-2 spike glycoprotein, due to its primary interaction with the human angiotensin-converting enzyme 2 (ACE2) cell-surface receptor, is considered as a potential target for drug development. Based on in silico screening followed by in vitro studies, here we report that the existing FDA-approved Bcr-Abl tyrosine kinase inhibitor, imatinib, inhibits SARS-CoV-2 with an IC50 of 130 nM. We provide evidence that although imatinib binds to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein with an affinity at micromolar, i.e., 2.32 ± 0.9 μM levels, imatinib does not directly inhibit the spike RBD:ACE2 interaction – suggesting a Bcr-Abl kinase-mediated fusion inhibition mechanism is responsible for the inhibitory action. This study utilizes in silico methodology followed by in vitro experimental validation to screen existing FDA-approved small molecule drugs specific to the RBD of the spike protein of SARS-CoV-2 to identify repurposable drugs targeting further clinical validation.

abstract:
  The rapid geographic expansion of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the infectious agent of Coronavirus Disease 2019 (COVID-19) pandemic, poses an immediate need for potent drugs. Enveloped viruses infect the host cell by cellular membrane fusion, a crucial mechanism required for virus replication. The SARS-CoV-2 spike glycoprotein, due to its primary interaction with the human angiotensin-converting enzyme 2 (ACE2) cell-surface receptor, is considered as a potential target for drug development. Based on in silico screening followed by in vitro studies, here we report that the existing FDA-approved Bcr-Abl tyrosine kinase inhibitor, imatinib, inhibits SARS-CoV-2 with an IC50 of 130 nM. We provide evidence that although imatinib binds to the receptor-binding domain (RBD) of SARS-CoV-2 spike protein with an affinity at micromolar, i.e., 2.32 ± 0.9 μM levels, imatinib does not directly inhibit the spike RBD:ACE2 interaction – suggesting a Bcr-Abl kinase-mediated fusion inhibition mechanism is responsible for the inhibitory action. We also show that imatinib inhibits other coronaviruses, SARS-CoV, and MERS-CoV via fusion inhibition. Based on promising in vitro results, we propose the Abl tyrosine kinase inhibitor (ATKI), imatinib, to be a viable repurposable drug against COVID-19.


id: cord-296399-vvbjulm9
author: Brinkmann, Constantin
date: 2017-12-07
url: https://www.ncbi.nlm.nih.gov/pubmed/29216247/
doi: 10.1371/journal.pone.0189073
title:
  The glycoprotein of vesicular stomatitis virus promotes release of virus-like particles from tetherin-positive cells

abstract:
  Vesicular stomatitis virus (VSV) release from infected cells is inhibited by the interferon (IFN)-inducible antiviral host cell factor tetherin (BST-2, CD317). However, several viruses encode tetherin antagonists and it is at present unknown whether residual VSV spread in tetherin-positive cells is also promoted by a virus-encoded tetherin antagonist. Here, we show that the viral glycoprotein (VSV-G) antagonizes tetherin in transfected cells, although with reduced efficiency as compared to the HIV-1 Vpu protein. Tetherin antagonism did not involve alteration of tetherin expression and was partially dependent on a GXXXG motif in the transmembrane domain of VSV-G. However, mutation of the GXXXG motif did not modulate tetherin sensitivity of infectious VSV. These results identify VSV-G as a tetherin antagonist in transfected cells but fail to provide evidence for a contribution of tetherin antagonism to viral spread.


id: cord-296466-hakaoo9i
author: Mäkelä, Anna R.
date: 2006-09-22
url: https://www.ncbi.nlm.nih.gov/pubmed/16997010/
doi: 10.1016/s0065-3527(06)68003-2
title:
  Baculovirus Display: A Multifunctional Technology for Gene Delivery and Eukaryotic Library Development

abstract:
  For over a decade, phage display has proven to be of immense value, allowing selection of a large variety of genes with novel functions from diverse libraries. However, the folding and modification requirements of complex proteins place a severe constraint on the type of protein that can be successfully displayed using this strategy, a restriction that could be resolved by similarly engineering a eukaryotic virus for display purposes. The quite recently established eukaryotic molecular biology tool, the baculovirus display vector system (BDVS), allows combination of genotype with phenotype and thereby enables presentation of eukaryotic proteins on the viral envelope or capsid. Data have shown that the baculovirus, Autographa californica multiple nucleopolyhedrovirus (AcMNPV), is a versatile tool for eukaryotic virus display. Insertion of heterologous peptides and/or proteins into the viral surface by utilizing the major envelope glycoprotein gp64, or foreign membrane‐derived counterparts, allows incorporation of the sequence of interest onto the surface of infected cells and virus particles. A number of strategies are being investigated in order to further develop the display capabilities of AcMNPV and improve the complexity of a library that may be accommodated. Numerous expression vectors for various approaches of surface display have already been developed. Further improvement of both insertion and selection strategies toward development of a refined tool for use in the creation of useful eukaryotic libraries is, however, needed. Here, the status of baculovirus display with respect to alteration of virus tropism, antigen presentation, transgene expression in mammalian cells, and development of eukaryotic libraries will be reviewed.


id: cord-299281-5z1xminb
title: Oligomerization of a membrane protein correlates with its retention in the Golgi complex
author: nan
date: 1993-09-02
url: https://www.ncbi.nlm.nih.gov/pubmed/8397214/
doi: nan
abstract:
  The first membrane-spanning domain (m1) of the M glycoprotein of avian coronavirus (formerly called E1) is sufficient to retain this protein in the cis-Golgi. When the membrane-spanning domain of a protein which is efficiently delivered to the plasma membrane (VSV G protein) is replaced with m1, the resulting chimera (Gm1) is retained in the Golgi (Swift, A. M., and C. E. Machamer. 1991. J. Cell Biol. 115:19-30). When assayed in sucrose gradients, we observed that Gm1 formed a large oligomer, and that much of this oligomer was SDS resistant and stayed near the top of the stacking gel of an SDS-polyacrylamide gel. The unusual stability of the oligomer allowed it to be detected easily. Gm1 mutants with single amino acid substitutions in the m1 domain that were retained in the Golgi complex formed SDS-resistant oligomers, whereas mutants that were rapidly released to the plasma membrane did not. Oligomerization was not detected immediately after synthesis of Gm1, but occurred gradually with a lag of approximately 10 min, suggesting that it is not merely aggregation of misfolded proteins. Furthermore, oligomerization did not occur under several conditions that block ER to Golgi transport. The lumenal domain was not required for oligomerization since another chimera (alpha m1G), where the lumenal domain of Gm1 was replaced by the alpha subunit of human chorionic gonadotropin, also formed an SDS-resistant oligomer, and was able to form hetero-oligomers with Gm1 as revealed by coprecipitation experiments. SDS resistance was conferred by the cytoplasmic tail of VSV G, because proteolytic digestion of the tail in microsomes containing Gm1 oligomers resulted in loss of SDS resistance, although the protease-treated material continued to migrate as a large oligomer on sucrose gradients. Interestingly, treatment of cells with cytochalasin D blocked formation of SDS-resistant (but not SDS- sensitive) oligomers. Our data suggest that SDS-resistant oligomers form as newly synthesized molecules of Gm1 arrive at the Golgi complex and may interact (directly or indirectly) with an actin-based cytoskeletal matrix. The oligomerization of Gm1 and other resident proteins could serve as a mechanism for their retention in the Golgi complex.


id: cord-316589-f1hq0xl5
author: Nagalo, Bolni Marius
date: 2020-08-19
txt: ./txt/cord-316589-f1hq0xl5.txt
cache: ./cache/cord-316589-f1hq0xl5.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1016/j.omto.2020.08.007
doi: 10.1016/j.omto.2020.08.007
summary:
  Our results indicate that high intrahepatic doses of VSV-FH did not result in any significant toxicity and were well tolerated by transgenic mice expressing the measles virus receptor CD46. Furthermore, single intratumoral treatments with VSV-FH yielded improved survival and complete tumor regressions in a proportion of mice in the Hep3B hepatocellular carcinoma model, but not in mice xenografted with BxPC3 pancreatic cancer cells. In this study, we evaluated whether treatment with oncolytic VSV-FH could trigger a potent cytotoxicity effect in HBPC cell lines in vitro and in vivo using animal models. Safety studies on intrahepatic or intratumoral injection of oncolytic vesicular stomatitis virus expressing interferon-beta in rodents and nonhuman primates High CD46 receptor density determines preferential killing of tumor cells by oncolytic measles virus Vesicular stomatitis virus expressing interferon-beta is oncolytic and promotes antitumor immune responses in a syngeneic murine model of non-small cell lung cancer

title:
  Oncolytic Virus With Attributes of Vesicular Stomatitis Virus and Measles Virus in Hepatobiliary and Pancreatic Cancers

abstract:
  Abstract Recombinant vesicular stomatitis virus (VSV)-fusion and hemagglutinin (FH) was developed by substituting the promiscuous VSV-G glycoprotein (G) gene in the backbone of VSV with genes encoding for the measles virus envelope proteins F and H. Hybrid VSV-FH exhibited a multifaceted mechanism of cancer-cell killing, and improved neurotolerability over parental VSV in preclinical studies. In this study, we evaluated VSV-FH in vitro and in vivo in models of hepatobiliary and pancreatic cancers. Our results indicate that high intrahepatic doses of VSV-FH did not result in any significant toxicity and were well tolerated by transgenic mice expressing the measles virus receptor CD46. Furthermore, single intratumoral treatments with VSV-FH yielded improved survival and complete tumor regressions in a proportion of mice in the Hep3B hepatocellular carcinoma model, but not in mice xenografted with BxPC3 pancreatic cancer cells. Our preliminary findings indicate that VSV-FH can induce potent oncolysis in hepatocellular and pancreatic cancer cell lines with concordant results in vivo in hepatocellular cancer and discordant in pancreatic cancer, without the VSV-mediated toxic effects previously observed in laboratory animals. Further study of VSV-FH as an oncolytic virotherapy is warranted in hepatocellular carcinoma and pancreatic cancer, to understand broader applicability and mechanisms of sensitivity and resistance.


id: cord-318587-ewvnkdr2
author: Steeds, Kimberley
date: 2020-08-31
url: https://www.ncbi.nlm.nih.gov/pubmed/32868837/
doi: 10.1038/s41598-020-71225-1
title:
  Pseudotyping of VSV with Ebola virus glycoprotein is superior to HIV-1 for the assessment of neutralising antibodies

abstract:
  Ebola virus (EBOV) is an enveloped, single-stranded RNA virus that can cause Ebola virus disease (EVD). It is thought that EVD survivors are protected against subsequent infection with EBOV and that neutralising antibodies to the viral surface glycoprotein (GP) are potential correlates of protection. Serological studies are vital to assess neutralising antibodies targeted to EBOV GP; however, handling of EBOV is limited to containment level 4 laboratories. Pseudotyped viruses can be used as alternatives to live viruses, which require high levels of bio-containment, in serological and viral entry assays. However, neutralisation capacity can differ among pseudotyped virus platforms. We evaluated the suitability of EBOV GP pseudotyped human immunodeficiency virus type 1 (HIV-1) and vesicular stomatitis virus (VSV) to measure the neutralising ability of plasma from EVD survivors, when compared to results from a live EBOV neutralisation assay. The sensitivity, specificity and correlation with live EBOV neutralisation were greater for the VSV-based pseudotyped virus system, which is particularly important when evaluating EBOV vaccine responses and immuno-therapeutics. Therefore, the EBOV GP pseudotyped VSV neutralisation assay reported here could be used to provide a better understanding of the putative correlates of protection against EBOV.


id: cord-318686-we6pveus
author: Ehlen, Lukas
date: 2016-05-04
txt: ./txt/cord-318686-we6pveus.txt
cache: ./cache/cord-318686-we6pveus.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/27142375/
doi: 10.1186/s12985-016-0531-5
summary:
  CONCLUSION: In the current study, we showed that newly established cell lines from the cotton rat can serve as host-specific in vitro models for viral infection experiments. The cotton rat (Sigmodon hispidus) is a unique example of a rodent species that is a well-established animal model to study viral pathogenesis and is also associated with a large range of zoonotic viruses in the wild [20] [21] [22] . To evaluate whether the broad viral susceptibility seen in both animalmodel and wild cotton rats was also reflected in in vitro cell culture models, we generated continuous cell lines from the respiratory and renal tracts of a cotton rat, and assessed their use for virus replication studies of known and potentially novel zoonotic viruses. In the work presented herein, we generated epithelial cell lines from the respiratory and renal tracts of a cotton rat due to its susceptibility to a broad range of human viruses, as well as the association of multiple important and emerging zoonotic viruses with this species.

title:
  Epithelial cell lines of the cotton rat (Sigmodon hispidus) are highly susceptible in vitro models to zoonotic Bunya-, Rhabdo-, and Flaviviruses

abstract:
  BACKGROUND: Small mammals such as bats and rodents have been increasingly recognized as reservoirs of novel potentially zoonotic pathogens. However, few in vitro model systems to date allow assessment of zoonotic viruses in a relevant host context. The cotton rat (Sigmodon hispidus) is a New World rodent species that has a long-standing history as an experimental animal model due to its unique susceptibility to human viruses. Furthermore, wild cotton rats are associated with a large variety of known or potentially zoonotic pathogens. METHODS: A method for the isolation and culture of airway epithelial cell lines recently developed for bats was applied for the generation of rodent airway and renal epithelial cell lines from the cotton rat. Continuous cell lines were characterized for their epithelial properties as well as for their interferon competence. Susceptibility to members of zoonotic Bunya-, Rhabdo-, and Flaviviridae, in particular Rift Valley fever virus (RVFV), vesicular stomatitis virus (VSV), West Nile virus (WNV), and tick-borne encephalitis virus (TBEV) was tested. Furthermore, novel arthropod-derived viruses belonging to the families Bunya-, Rhabdo-, and Mesoniviridae were tested. RESULTS: We successfully established airway and kidney epithelial cell lines from the cotton rat, and characterized their epithelial properties. Cells were shown to be interferon-competent. Viral infection assays showed high-titre viral replication of RVFV, VSV, WNV, and TBEV, as well as production of infectious virus particles. No viral replication was observed for novel arthropod-derived members of the Bunya-, Rhabdo-, and Mesoniviridae families in these cell lines. CONCLUSION: In the current study, we showed that newly established cell lines from the cotton rat can serve as host-specific in vitro models for viral infection experiments. These cell lines may also serve as novel tools for virus isolation, as well as for the investigation of virus-host interactions in a relevant host species.


id: cord-324674-yd7idp90
author: Zhang, Chengfei
date: 2018-08-22
txt: ./txt/cord-324674-yd7idp90.txt
cache: ./cache/cord-324674-yd7idp90.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1093/jmcb/mjy045
doi: 10.1093/jmcb/mjy045
summary:
  ADP/P2Y 13 -mediated protection against viral infection operates by suppressing the expression of exchange protein activated by cAMP 1 (EPAC1), which is an alternative key intracellular sensor for cAMP. To our surprise, the RNA replication of VSV in ADP-treated RAW264.7 cells was reduced significantly in a time-( Figure 2D ) and concentration-( Figure 2E ) dependent manner. To explore the key receptors involved in ADP-mediated antiviral activities, we detected the expression of P2Y 1 , P2Y 12 , and P2Y 13 after VSV infection. ADP/P2Y 13 restricts viral replication by inhibiting cAMP signaling Type I IFN plays pivotal roles in fighting against the invaded virus, so we tested whether it was involved in ADP/P2Y 13mediated antiviral activities. As shown in Figure 7A , when infected RAW264.7 cells with VSV, NDV, and HSV-1, RNA expression of EPAC1 was increased significantly.

title:
  IFN-stimulated P2Y(13) protects mice from viral infection by suppressing the cAMP/EPAC1 signaling pathway

abstract:
  Among the most important sensors of extracellular danger signals, purinergic receptors have been demonstrated to play crucial roles in host defense against infection. However, the function of P2 receptors in viral infection has been little explored. Here we demonstrated that P2Y(13) and its ligand ADP play an important role in protecting hosts from viral infections. First, we demonstrate that P2Y(13), as a typical interferon-stimulated gene, is induced together with extracellular ADP during viral infection. Most importantly, extracellular ADP restricts the replication of different kinds of viruses, including vesicular stomatitis virus, Newcastle disease virus, herpes simplex virus 1, and murine leukemia virus. This kind of protection is dependent on P2Y(13) but not P2Y(1) or P2Y(12), which are also considered as receptors for ADP. Furthermore, cyclic adenosine monophosphate and EPAC1 are downregulated by extracellular ADP through the P2Y(13)-coupled Gi alpha subunit. Accordingly, inhibition or deletion of EPAC1 significantly eliminates ADP/P2Y(13)-mediated antiviral activities. Taken together, our results show that P2Y(13) and ADP play pivotal roles in the clearance of invaded virus and have the potential as antiviral targets.


id: cord-326013-5i35zdmv
author: Carpinteiro, Alexander
date: 2020-10-29
txt: ./txt/cord-326013-5i35zdmv.txt
cache: ./cache/cord-326013-5i35zdmv.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/33163980/
doi: 10.1016/j.xcrm.2020.100142
summary:
  The data justify clinical studies investigating whether amitriptyline, a safe drug used clinically for almost 60 years, or other antidepressants that functionally block acid sphingomyelinase prevent SARS-CoV-2 infection. Pretreatment of the cells with 5, 10, 20, or 25 µM amitriptyline prevented the activation of acid sphingomyelinase and the release of ceramide upon infection with pp-VSV-SARS-CoV-2 spike for 30 min (Fig. 3B, Fig. 4A ). Treating Vero cells with neutralizing antibodies to spike or with recombinant ACE2 protein prevented the activation of acid sphingomyelinase and the release of ceramide upon infection with pp-VSV-SARS-CoV-2 spike (Fig. 3B, Fig. 4A Amitriptyline and other drugs with similar structure and properties have been clinically used for many years (since 1962) to treat patients with depressive disorder. Best results were obtained with venlafaxin, fluoxetine, escitalopram and mirtazipine, drugs that were also shown in the present study to inhibit acid sphingomyelinase and ceramide release upon pp-VSV-SARS-CoV-2 spike infection.

title:
  Pharmacological inhibition of acid sphingomyelinase prevents uptake of SARS-CoV-2 by epithelial cells

abstract:
  The acid sphingomyelinase/ceramide system plays an important role in bacterial and viral infections. Here we report that either pharmacological inhibition of acid sphingomyelinase with amitriptyline, imipramine, fluoxetine, sertraline, escitalopram, or maprotiline, or genetic downregulation of the enzyme prevents infection of cultured cells or freshy isolated human nasal epithelial cells with SARS-CoV-2 or pseudoviral pp-VSV-SARS-CoV-2 particles expressing spike, a bona fide system mimicking SARS-CoV-2 infection. Infection activates acid sphingomyelinase and triggers a release of ceramide on the cell surface. Neutralization or consumption of surface ceramide reduces infection with pp-VSV-SARS-CoV-2 spike. Treating volunteers with a low dose of amitriptyline prevents infection of freshly isolated nasal epithelial cells with pp-VSV-SARS-CoV-2 spike. The data justify clinical studies investigating whether amitriptyline, a safe drug used clinically for almost 60 years, or other antidepressants that functionally block acid sphingomyelinase prevent SARS-CoV-2 infection.


id: cord-327199-ggomuomb
title: Cellular Proteins Associated with the Interior and Exterior of Vesicular Stomatitis Virus Virions
author: Moerdyk-Schauwecker, Megan
date: 2014-08-08
url: https://www.ncbi.nlm.nih.gov/pubmed/25105980/
doi: 10.1371/journal.pone.0104688
abstract:
  Virus particles (virions) often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact (“whole”) virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i) whole VSV virions; and ii) whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y), was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.


id: cord-339854-scb7pz87
title: The synthetic futures of vesicular stomatitis virus
author: Overend, Christopher
date: 2012-07-11
txt: ./txt/cord-339854-scb7pz87.txt
cache: ./cache/cord-339854-scb7pz87.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1016/j.tibtech.2012.06.002
doi: 10.1016/j.tibtech.2012.06.002
abstract: nan
summary:
  Vesicular stomatitis virus (VSV) is one of the most promising viruses for engineering vaccines and oncolytic therapies [2] . Of particular interest is a study in which VSV expressing the H5 antigen from highly pathogenic avian influenza induced sterilizing immunity against heterologous challenge in mice [4] . This demonstrates the safety and efficacy potential of VSV when live virus vaccination would otherwise be contraindicated. Even more promising, recombinant VSV expressing a secreted form of a virulence factor protein for Yersinia pestis, LcrV, induced high levels of LcrV-specific antibodies, protecting 90% of the mice challenged with 10 LD 50 [6] . Vesicular stomatitis virus-based Ebola vaccine is well-tolerated and protects immunocompromised nonhuman primates Potent vesicular stomatitis virus-based avian influenza vaccines provide long-term sterilizing immunity against heterologous challenge Heterologous boosting of recombinant adenoviral prime immunization with a novel vesicular stomatitis virus-vectored tuberculosis vaccine SARS vaccine based on a replicationdefective recombinant vesicular stomatitis virus is more potent than one based on a replication-competent vector


id: cord-345299-4k7qymqd
title: Several FDA-approved drugs effectively inhibit SARS-CoV-2 infection in vitro
author: Xiong, Hua-Long
date: 2020-06-05
txt: ./txt/cord-345299-4k7qymqd.txt
cache: ./cache/cord-345299-4k7qymqd.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.06.05.135996
doi: 10.1101/2020.06.05.135996
summary:
  To identify drugs that are potentially used for the treatment of COVID-19, the potency of 1403 FDA-approved drugs were evaluated using a robust pseudovirus assay and the candidates were further confirmed by authentic SARS-CoV-2 assay. Four compounds, Clomiphene (citrate), Vortioxetine, Vortioxetine (hydrobromide) and Asenapine (hydrochloride), showed potent inhibitory effects in both pseudovirus and authentic virus assay. In this study, the anti-SARS-Cov-2 potentiality of 1403 FDA approved drugs were quantitatively evaluated by the pseudovirus-based assay. In the second round of screening, inhibition of VSV-SARS-CoV-2-Sdel18 virus infection and cell cytotoxicity were both detected (Supplementary Figure 1) . The robust assay based on VSV-SARS-CoV-2-Sdel18 pseudovirus screened out the potential drugs with high efficiency, then the inhibitory effect was confirmed by authentic SARS-CoV-2 assay. The relative value or inhibition rate of candidate drugs were calculated according to the decrease of GFP positive cell number (for pseudovirus-based assay) or cytopathic effect (for authentic SARS-CoV-2-based assay).

abstract:
  To identify drugs that are potentially used for the treatment of COVID-19, the potency of 1403 FDA-approved drugs were evaluated using a robust pseudovirus assay and the candidates were further confirmed by authentic SARS-CoV-2 assay. Four compounds, Clomiphene (citrate), Vortioxetine, Vortioxetine (hydrobromide) and Asenapine (hydrochloride), showed potent inhibitory effects in both pseudovirus and authentic virus assay. The combination of Clomiphene (citrate), Vortioxetine and Asenapine (hydrochloride) is much more potent than used alone, with IC50 of 0.34 μM.


id: cord-346554-a98pjtxs
title: Inhibitory effects of bee venom and its components against viruses in vitro and in vivo
author: Uddin, Md Bashir
date: 2016-11-26
url: https://www.ncbi.nlm.nih.gov/pubmed/27888461/
doi: 10.1007/s12275-016-6376-1
abstract:
  Bee venom (BV) from honey bee (Apis Melifera L.) contains at least 18 pharmacologically active components including melittin (MLT), phospholipase A(2) (PLA(2)), and apamin etc. BV is safe for human treatments dose dependently and proven to possess different healing properties including antibacterial and antiparasitidal properties. Nevertheless, antiviral properties of BV have not well investigated. Hence, we identified the potential antiviral properties of BV and its component against a broad panel of viruses. Co-incubation of non-cytotoxic amounts of BV and MLT, the main component of BV, significantly inhibited the replication of enveloped viruses such as Influenza A virus (PR8), Vesicular Stomatitis Virus (VSV), Respiratory Syncytial Virus (RSV), and Herpes Simplex Virus (HSV). Additionally, BV and MLT also inhibited the replication of non-enveloped viruses such as Enterovirus-71 (EV-71) and Coxsackie Virus (H3). Such antiviral properties were mainly explained by virucidal mechanism. Moreover, MLT protected mice which were challenged with lethal doses of pathogenic influenza A H1N1 viruses. Therefore, these results provides the evidence that BV and MLT could be a potential source as a promising antiviral agent, especially to develop as a broad spectrum antiviral agent.


id: cord-346777-zmmnn9b2
author: Lester, Sandra
date: 2019-09-11
url: https://doi.org/10.1099/acmi.0.000057
doi: 10.1099/acmi.0.000057
title:
  Middle East respiratory coronavirus (MERS-CoV) spike (S) protein vesicular stomatitis virus pseudoparticle neutralization assays offer a reliable alternative to the conventional neutralization assay in human seroepidemiological studies

abstract:
  Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel zoonotic coronavirus that was identified in 2012. MERS-CoV infection in humans can result in an acute, severe respiratory disease and in some cases multi-organ failure; the global mortality rate is approximately 35 %. The MERS-CoV spike (S) protein is a major target for neutralizing antibodies in infected patients. The MERS-CoV microneutralization test (MNt) is the gold standard method for demonstrating prior infection. However, this method requires the use of live MERS-CoV in biosafety level 3 (BSL-3) containment. The present work describes the generation and validation of S protein-bearing vesicular stomatitis virus (VSV) pseudotype particles (VSV-MERS-CoV-S) in which the VSV glycoprotein G gene has been replaced by the luciferase reporter gene, followed by the establishment of a pseudoparticle-based neutralization test to detect MERS-CoV neutralizing antibodies under BSL-2 conditions. Using a panel of human sera from confirmed MERS-CoV patients, the VSV-MERS-CoV particle neutralization assay produced results that were highly comparable to those of the microneutralization test using live MERS-CoV. The results suggest that the VSV-MERS-CoV-S pseudotype neutralization assay offers a highly specific, sensitive and safer alternative method to detect MERS-CoV neutralizing antibodies in human sera.


id: cord-351881-qea4b0i5
author: Eck, Melanie
date: 2016-02-19
url: https://www.ncbi.nlm.nih.gov/pubmed/26895704/
doi: 10.1186/s13567-016-0318-0
title:
  Virus replicon particles expressing porcine reproductive and respiratory syndrome virus proteins elicit immune priming but do not confer protection from viremia in pigs

abstract:
  Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of one of the most devastating and economically significant viral disease of pigs worldwide. The vaccines currently available on the market elicit only limited protection. Recombinant vesicular stomatitis virus (VSV) replicon particles (VRP) have been used successfully to induce protection against influenza A virus (IAV) in chickens and bluetongue virus in sheep. In this study, VSV VRP expressing the PRRSV envelope proteins GP5, M, GP4, GP3, GP2 and the nucleocapsid protein N, individually or in combination, were generated and evaluated as a potential vector vaccine against PRRSV infection. High level expression of the recombinant PRRSV proteins was demonstrated in cell culture. However, none of the PRRSV antigens expressed from VRP, with the exception of the N protein, did induce any detectable antibody response in pigs before challenge infection with PRRSV. After challenge however, the antibody responses against GP5, GP4 and GP3 appeared in average 2 weeks earlier than in pigs vaccinated with the empty control VRP. No reduction of viremia was observed in the vaccinated group compared with the control group. When pigs were co-vaccinated with VRP expressing IAV antigens and VRP expressing PRRSV glycoproteins, only antibody responses to the IAV antigens were detectable. These data show that the VSV replicon vector can induce immune responses to heterologous proteins in pigs, but that the PRRSV envelope proteins expressed from VSV VRP are poorly immunogenic. Nevertheless, they prime the immune system for significantly earlier B-cell responses following PRRSV challenge infection.


