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Serological Relationships between Different Strains of Vesicular Stomatis Virus

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Abstract

SUMMARY The five vesicular stomatitis viruses Indiana, New Jersey, Cocal, Argentina and Brazil have been examined for cross-relationships by complement fixing and neutralization tests. With the exception of the Cocal and Argentina strains, intact virus particles showed little cross-reaction in either test. However, the infective skeleton-like structure produced by uncoating the virus with Tween-ether or Nonidet, and the infective ribonucleoprotein (RNP) produced by deoxycholate disruption of the virus, were neutralized by either homo- or heterotypic antisera and showed considerable cross-reactivity in the complement fixation test. Experi- ments with the isolated sub-units of the virus indicated that the major part of the cross-reactivity was associated with the RNP. The morphologically similar Piry and Chandipura viruses appeared to be unrelated to the vesicular stomatitis group in both complement fixation and neutralization tests.

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... Based on these viruses' natural history, different cell tropisms are expected, with Chandipura virus to be more neurotropic. Another advantage with the use of these new envelopes is that they are immunologically non-cross reactive with VSV-G and thus permitting their use in vivo in a sequential manner as an alternative to VSV-G pseudotyped vectors for boosting gene delivery and dosage as becomes necessary (Brun et al., 1995;Cartwright and Brown, 1972). Here our results show that lentiviral vectors could be efficiently pseudotyped with the envelopes from both these viruses, there are tropism and transduction efficiency differences based on cell types, that the new envelopes display stability for freeze thawing and finally, they are relatively more resistant to exposure to human serum. ...
... Here in an effort to identify alternative heterologous viral envelopes for vector pseudotyping, we evaluated two related viral envelopes from Chandipura and Piry viruses that belong to the same Vesiculovirus genus like VSV. While both these viruses are phylogenetically related to VSV, they are serologically distinct and with different cell tropisms (Brun et al., 1995;Cartwright and Brown, 1972). ...
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While the envelope glycoprotein of vesicular stomatitis virus (VSV-G) is widely used for pseudotyping of lentiviral vectors, sub-optimal gene transfer into certain cell types and its sensitivity to inactivation by human complement hinders its broader applications. To find alternative candidates, here we evaluated two serologically distinct novel viral envelopes derived from Chandipura (CNV-G) and Piry (PRV-G) vesiculoviruses. Both permitted generation of high titer psuedotyped lentiviral vectors with a capacity for high efficiency gene transfer into various cell types from different species. In human lymphoid and hematopoietic stem cells, their transduction efficiency was significantly lower than that of VSV-G. However, both novel envelopes were found to be more resistant to inactivation by human serum complement compared to VSV-G. Thus CNV-G and PRV-G envelopes can be harnessed for multiple uses in the future based on the cell type that needs to be gene transduced and possibly for in vivo gene transfer.
... Vesicular stomatitis (VS) is a viral disease caused by a Rhabdovirus that infects cattle, horses, and swine [1]. VS viruses (VSV) are classified by serotypes, Indiana (VSV-IN) and New Jersey (VSV-NJ), based on the distinct neutralizing antibodies generated in infected animals [2,3]. Clinical signs include vesicular lesions in the gums, tongue, oro-nasal mucosa, teats, and coronary bands [4,5]. ...
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Vesicular stomatitis virus (VSV) primarily infects livestock and is transmitted by direct contact and vectored by Culicoides midges (Diptera: Ceratopogonidae). Endemic to Central and South America, specific VSV lineages spread northward out of endemic regions of Mexico and into the U.S. sporadically every five to ten years. In 2012, a monophyletic epidemic lineage 1.1 successfully spread northward into the U.S. In contrast, the closest endemic ancestor, lineage 1.2, remained circulating exclusively in endemic regions in Mexico. It is not clear what roles virus-animal interactions and/or virus-vector interactions play in the ability of specific viral lineages to escape endemic regions in Mexico and successfully cause outbreaks in the U.S., nor the genetic basis for such incursions. Whole-genome sequencing of epidemic VSV 1.1 and endemic VSV 1.2 revealed significant differences in just seven amino acids. Previous studies in swine showed that VSV 1.1 was more virulent than VSV 1.2. Here, we compared the efficiency of these two viral lineages to infect the vector Culicoides sonorensis (Wirth and Jones) and disseminate to salivary glands for subsequent transmission. Our results showed that midges orally infected with the epidemic VSV 1.1 lineage had significantly higher infection dissemination rates compared to those infected with the endemic VSV 1.2 lineage. Thus, in addition to affecting virus-animal interactions, as seen with higher virulence in pigs, small genetic changes may also affect virus-vector interactions, contributing to the ability of specific viral lineages to escape endemic regions via vector-borne transmission.
... In spite of the antigenic differences between VSV serotypes, variable levels of cross-neutralization are observed among VSIV-1, 2 e 3 (PAUSZEK et al., 2011). In contrast, crossneutralization between VSIV and VSNJV occurs at very low levels due to their antigenic differences (CARTWRIGHT & BROWN, 1972). Positivity to VSNJV is probably to this cross-reactivity because VSNJV is considered exotic in Brazil (OIE, 2010;PANAFTOSA, 2015). ...
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Vesicular stomatitis virus (VSV) is the agent of a vesicular disease that affects many animal species and may be clinically confounded with foot-and-mouth disease in ruminant and swine. Horses are especially susceptible to VSV and may serve as sentinels for virus circulation. The present study investigated the presence of neutralizing antibodies against VSV Indiana III (VSIV-3) in serum samples of 3,626 horses from six states in three Brazilian regions: Southern (RS, n = 1,011), Midwest (GO/DF, n = 1,767) and Northeast (PB, PE, RN and CE, n = 848) collected between 2013 and 2014. Neutralizing antibodies against VSIV-3 (titers ≥40) were detected in 641 samples (positivity of 17.7%; CI95%:16.5-19.0%), being 317 samples from CE (87.3%; CI95%: 83.4-90.5 %); 109 from RN (65.7%; CI95%: 57.8 -72.7%); 124 from PB (45.4%; CI95%: 39.4-51.5%); 78 from GO/DF (4.4%; CI95%: 3.5-5.5%) and nine samples of RS (0.9%; CI95%: 0.4-1.7%). Several samples from the Northeast and Midwest harbored high neutralizing titers, indicating a recent exposure to the virus. In contrast, samples from RS had low titers, possibly due to a past remote exposure. Several positive samples presented neutralizing activity against other VSV serotypes (Indiana I and New Jersey), yet in lower titers, indicating the specificity of the response to VSIV-3. These results demonstrated a relatively recent circulation of VSIV-3 in northeastern Brazilian States, confirming clinical findings and demonstrating the sanitary importance of this infection.
... VSVNJ and VSVInd do not share appreciable Finally, the presence of common 5'-terminal sequence homologies. Nevertheless, both the se-sequences of the mRNA's and the leader RNAs rotypes have a cross-reacting group-specific an-of VSVNJ and VSVInd indicate that these setigen in the ribonucleoprotein cores (9). This quences may have been conserved if these isoapparent contradiction has not yet been re-lates have evolved from a common ancestor. ...
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The New Jersey serotype of vesicular stomatitis virus (VSV) was able to synthesize a small RNA (leader RNA) approximately 70 bases in length similar to the leader RNA synthesized in vitro by the genetically distinct Indiana serotype of VSV. Also, the New Jersey leader RNA contained the same 5'-terminal sequence, ppA-C-G, as the Indiana leader RNA and had a very similar base composition, with 42% AMP, 16% CMP, 18.6% GMP, and 23.4% UMP. The 3'-terminal sequence of the VSV New Jersey genome RNA was detemined and found to contain the sequence- Py-G-UOH, again the same as that of the Indiana serotype of VSV. Evidence that the New Jersey leader RNA is transcribed from the 3' end of the genome RNA was obtained from the fact that it can protect the 3'-terminal base of [3H]borohydride-labeled New Jersey genome RNA from RNase digestion. Although the New Jersey and Indiana leader RNAs were similar in many respects, they were unable to form RNase-resistant hybrids when annealed to heterologous genome RNA.
... These relationships were initially characterized by two-way serological cross-reactions (R values) based on either complement fixation or neutralization tests using hyperimmune sera generated in guinea pigs [1]. Complement fixation tests allow for more cross-reactions, since they are based on more conserved viral proteins (e.g., N) [1,5,13]. Neutralization tests are serotype-specific and reflect differences in the sequence of the viral glycoprotein, since neutralizing epitopes are located exclusively on this protein [4,15,17]. ...
Article
Vesicular stomatitis (VS) viruses have been classified into two serotypes: New Jersey (VSNJV) and Indiana (VSIV). Here, we have characterized field isolates causing vesicular stomatitis in Brazil and Argentina over a 35-year span. Cluster analysis based on either serological relatedness, as inferred from virus neutralization and complement fixation assays, or nucleotide sequences of two separate genes (phosphoprotein or glycoprotein) grouped the field isolates into two distinct monophyletic groups within the Indiana serogroup. One group included seven viruses from Brazil and Argentina that were serologically classified as Indiana-2 and Cocal virus (COCV). The other group contained three viruses from Brazil that were serologically classified as Indiana-3 and the prototype of this group, Alagoas virus (VSAV). Interestingly, two vesiculoviruses that were isolated from insects but do not cause disease in animals, one from Brazil (Maraba virus; MARAV) and the other from Colombia (CoAr 171638), grouped into two separate genetic lineages within the Indiana serotype. Our data provide support for the classification of viruses causing clinical VS in livestock in Brazil and Argentina into two distinct groups: Indiana-2 (VSIV-2) and Indiana-3 (VSIV-3). We suggest using nomenclature for these viruses that includes the serotype, year and place of occurrence, and affected host. This nomenclature is consistent with that currently utilized to describe field isolates of VSNJV or VSIV in scientific literature.
... Two different serotypes of VSV, VSV-Indiana (VSV Ind ) and VSV-New Jersey (VSV NJ ), show 50% amino acid homologies in glycoprotein G (4). Antibodies raised against one serotype of VSV do not neutralize the other serotype of VSV (5). Accordingly, other investigators have used VSV Ind with its own surface glycoprotein G and VSV Ind carrying the G gene from other serotypes as a vaccine vector (6,7). ...
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Unlabelled: To take advantage of live recombinant vesicular stomatitis viruses (rVSVs) as vaccine vectors for their high yield and for their induction of strong and long-lasting immune responses, it is necessary to make live vaccine vectors safe for use without losing their immunogenicity. We have generated safer and highly efficient recombinant VSV vaccine vectors by combining the M51R mutation in the M gene of serotype VSV-Indiana (VSVInd) with a temperature-sensitive mutation (tsO23) of the VSVInd Orsay strain. In addition, we have generated two new serotype VSV-New Jersey (VSVNJ) vaccine vectors by combining M48R and M51R mutations with G22E and L110F mutations in the M gene, rVSVNJ(G22E M48R M51R) [rVSVNJ(GMM)] and VSVNJ(G22E M48R M51R L110F) [rVSVNJ(GMML)]. The combined mutations G21E, M51R, and L111F in the M protein of VSVInd significantly reduced the burst size of the virus by up to 10,000-fold at 37°C without affecting the level of protein expression. BHK21 cells and SH-SY5Y human neuroblastoma cells infected with rVSVInd(GML), rVSVNJ(GMM), and rVSVNJ(GMML) showed significantly reduced cytopathic effects in vitro at 37°C, and mice injected with 1 million infectious virus particles of these mutants into the brain showed no neurological dysfunctions or any other adverse effects. In order to increase the stability of the temperature-sensitive mutant, we have replaced the phenylalanine with alanine. This will change all three nucleotides from UUG (leucine) to GCA (alanine). The resulting L111A mutant showed the temperature-sensitive phenotype of rVSVInd(GML) and increased stability. Twenty consecutive passages of rVSVInd(GML) with an L111A mutation did not convert back to leucine (UUG) at position 111 in the M protein gene. Importance: Recombinant vesicular stomatitis viruses as live vaccine vectors are very effective in expressing foreign genes and inducing adaptive T cell and B cell immune responses. As with any other live viruses in humans or animals, the use of live rVSVs as vaccine vectors demands the utmost safety. Our strategy to attenuate rVSVInd by utilizing a temperature-sensitive assembly-defective mutation of L111A and combining it with an M51R mutation in the M protein of rVSVInd significantly reduced the pathogenicity of the virus while maintaining highly effective virus production. We believe our new temperature-sensitive M gene mutant of rVSVInd(GML) and M gene mutants of rVSVNJ(GMM) and rVSVNJ(GMML) add excellent vaccine vectors to the pool of live viral vectors.
... Existen dos serotipos virales Indiana (I) y New Jersey (NJ) los cuales son clasificados basados en los anticuerpos neutralizantes contra la glicoproteína G. Lo anterior es debido a que solamente hay un 50% de similitud a nivel de aminoácidos entre la glicoproteína de I y NJ (House et al., 2003). Del serotipo I se conocen tres subtipos I1 ó clásico, I2 ó Cocal e I3 ó Alagoas (Cartwright et al., 1972). En Colombia se presentan los serotipos NJ e I1 . ...
... For rhabdoviruses, the viral G protein is the only viral protein on the external surface of virions, and it mediates attachment to cellular receptors. In the well-studied mammalian rhabdovirus vesicular stomatitis virus (VSV), two major serotypes, New Jersey (VSNJV) and Indiana (VSIV), have been described based on neutralizing antibodies to the surface glycoprotein (G) (Cartwright & Brown 1972 ), and these serotypes differ in virulence for swine (Bridges, McCluskey, Salman, Hurd & Dick 1997). By using recombinant VSV viruses that differed only in their G genes, Martinez, Rodriguez, Jimenez, Pauszek & Wertz (2003) demonstrated that the G protein was a determinant of VSV virulence in this natural host. ...
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Previously, we demonstrated that a representative M genogroup type strain of infectious haematopoietic necrosis virus (IHNV) from rainbow trout grows well in rainbow trout-derived RTG-2 cells, but a U genogroup type strain from sockeye salmon has restricted growth, associated with reduced genome replication and mRNA transcription. Here, we analysed further the mechanisms for this growth restriction of U-type IHNV in RTG-2 cells, using strategies that assessed differences in viral genes, host immune regulation and phosphorylation. To determine whether the viral glycoprotein (G) or non-virion (NV) protein was responsible for the growth restriction, four recombinant IHNV viruses were generated in which the G gene of an infectious IHNV clone was replaced by the G gene of U- or M-type IHNV and the NV gene was replaced by NV of U- or M-type IHNV. There was no significant difference in the growth of these recombinants in RTG-2 cells, indicating that G and NV proteins are not major factors responsible for the differential growth of the U- and M-type strains. Poly I:C pretreatment of RTG-2 cells suppressed the growth of both U- and M-type IHNV, although the M virus continued to replicate at a reduced level. Both viruses induced type 1 interferon (IFN1) and the IFN1 stimulated gene Mx1, but the expression levels in M-infected cells were significantly higher than in U-infected cells and an inhibitor of the IFN1-inducible protein kinase PKR, 2-aminopurine (2-AP), did not affect the growth of U- or M-type IHNV in RTG-2 cells. These data did not indicate a role for the IFN1 system in the restricted growth of U-type IHNV in RTG-2 cells. Prediction of kinase-specific phosphorylation sites in the viral phosphoprotein (P) using the NetPhosK program revealed differences between U- and M-type P genes at five phosphorylation sites. Pretreatment of RTG-2 cells with a PKC inhibitor or a p38MAPK inhibitor did not affect the growth of the U- and M-type viruses. However, 100 μm of the casein kinase II (CKII) inhibitor, 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole (DRB), reduced the titre of the U type 8.3-fold at 24 h post-infection. In contrast, 100 μm of the CKII inhibitor reduced the titre of the M type only 1.3-fold at 48 h post-infection. Our data suggest that the different growth of U- and M-type IHNV in RTG-2 cells may be linked to a differential requirement for cellular protein kinases such as CKII for their growth.
... In spite of the antigenic differences between VSV serotypes, variable levels of cross-neutralization are observed among VSIV-1, 2 e 3 (PAUSZEK et al., 2011). In contrast, cross-neutralization between VSIV and VSNJV occurs in very low levels due to their antigenic differences (CARTWRIGHT & BROWN, 1972). The positivity to VSNJV is probably to this cross-reactivity since VSNJV is considered exotic in Brazil PANAFTOSA, 2015). ...
... Genetic fitness studies designed to investigate evolutionary pressure on the genome of VSV alternating between insect and mammalian cellular environments suggest that the stability of field populations is not due to the need of the virus to constrain adaptation between host cell types [38]. VS viruses are classified by serotypes, which are similar in size and morphology, but generate distinct neutralizing antibodies in infected animals [39,40]. There are two distinct serotypes of VSV: Indiana (VSV-IN) and New Jersey (VSV-NJ), with the latter causing the majority of outbreaks in the U.S. [2,7,41,42]. ...
... This induced robust adaptive immune responses against the inserted gene products [20,21]. Antibodies raised against one serotype of rVSV did not neutralize the other serotype [22]. Therefore, the impact of anti-vector antibodies arising after the priming immunization was avoided or minimized, resulting in a stronger immune response to the inserted immunogen. ...
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The development of safe and effective vaccines to prevent SARS-CoV-2 infections remains an urgent priority worldwide. We have used a recombinant vesicular stomatitis virus (rVSV)-based prime-boost immunization strategy to develop an effective COVID-19 vaccine candidate. We have constructed VSV genomes carrying exogenous genes resulting in the production of avirulent rVSV carrying the full-length spike protein (S F ), the S1 subunit, or the receptor-binding domain (RBD) plus envelope (E) protein of SARS-CoV-2. Adding the honeybee melittin signal peptide (msp) to the N-terminus enhanced the protein expression, and adding the VSV G protein transmembrane domain and the cytoplasmic tail (Gtc) enhanced protein incorporation into pseudotype VSV. All rVSVs expressed three different forms of SARS-CoV-2 spike proteins, but chimeras with VSV-Gtc demonstrated the highest rVSV-associated expression. In immunized mice, rVSV with chimeric S protein-Gtc derivatives induced the highest level of potent neutralizing antibodies and T cell responses, and rVSV harboring the full-length msp-S F -Gtc proved to be the superior immunogen. More importantly, rVSV-msp-S F -Gtc vaccinated animals were completely protected from a subsequent SARS-CoV-2 challenge. Overall, we have developed an efficient strategy to induce a protective response in SARS-CoV-2 challenged immunized mice. Vaccination with our rVSV-based vector may be an effective solution in the global fight against COVID-19.
... Genetic fitness studies designed to investigate evolutionary pressure on the genome of VSV alternating between insect and mammalian cellular environments suggest that the stability of field populations is not due to the need of the virus to constrain adaptation between host cell types [38]. VS viruses are classified by serotypes, which are similar in size and morphology, but generate distinct neutralizing antibodies in infected animals [39,40]. There are two distinct serotypes of VSV: Indiana (VSV-IN) and New Jersey (VSV-NJ), with the latter causing the majority of outbreaks in the U.S. [2,7,41,42]. ...
Article
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Vesicular stomatitis (VS) is a viral disease of veterinary importance, enzootic in tropical and subtropical regions of the Americas. In the U.S., VS produces devastating economic losses, particularly in the southwestern states where the outbreaks display an occurrence pattern of 10-year intervals. To date, the mechanisms of the geographic spread and maintenance cycles during epizootics remain unclear. This is due, in part, to the fact that VS epidemiology has a complex of variables to consider, including a broad range of vertebrate hosts, multiple routes of transmission, and an extensive diversity of suspected vector species acting as both mechanical and biological vectors. Infection and viral progression within vector species are highly influenced by virus serotype, as well as environmental factors, including temperature and seasonality; however, the mechanisms of viral transmission, including non-conventional pathways, are yet to be fully studied. Here, we review VS epidemiology and transmission mechanisms, with comparisons of transmission evidence for the four most incriminated hematophagous dipteran taxa: Aedes mosquitoes, Lutzomyia sand flies, Simulium black flies, and Culicoides biting midges.
... VSV Ind and VSV NJ show 50% amino acid homology in their glycoprotein G [13]. However, antibodies raised against VSV Ind do not neutralize VSV NJ [14]. Therefore, other investigators have used VSV Ind as a vaccine vector in which the glycoprotein gene of VSV Ind was replaced with that of VSV NJ to minimize the problems arising from this immune response against the same viral vectors [8,15]. ...
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Recently, we developed attenuated VSV vectors by introducing temperature sensitive (ts) mutations in the M gene in both VSV Indiana and New Jersey serotypes. The newly generated M gene mutants of rVSV vectors are rVSVInd(GML) with mutations of G21E, M51R, and L111F, rVSVNJ(GMM) with mutations of G22E, M48R, and M51R, and rVSVNJ(GMML) with mutations of G22E, M48R, M51R, and L110F. Our purpose was to examine the immunogenicity of the new ts M gene mutant of rVSVInd and attenuated M gene mutants ofrVSVNJas vaccine vectors against HIV-1 proteins. We generated attenuated rVSVs carrying HIV-1 gag, pol, and env genes. We immunized mice with various prime-boost vaccination regimens. CD8+ T cell responses and humoral immune responses in the vaccinated mice were examined. Priming with rVSVInd(GML)-gag, pol, or env gene of HIV-1 and boosting with rVSVNJ(GMM)-gag, pol, or env gene or rVSVNJ(GMML)-gag, pol, or env gene induced the strongest CD8+ cytotoxic T cell responses against the HIV-1 Gag, RT, and Env proteins. The same vaccination regimen also induced strong humoral immune responses against the HIV-1 Gag and Env proteins. We conclude that rVSVInd(GML) priming followed by rVSVNJ(GMM) boosting is the best vaccination regimen for optimum B cell and T cell adaptive immune responses against inserted foreign gene products when the newly attenuated rVSVInd and rVSVNJ are used.
Article
This overview attempts to classify and bring more order to the vast information available on animal virus replicative processes. Figure 1 presents the basic replicative pathways. The polarity convention for strandedness in the figure and text allows comparisons to be made across the virus classes, whereas vRNA, cRNA and cDNA designations do not. The IUR concept is obviously useful in classification, and mRNA with negative polarity is needed to accurately depict the symmetrical transcriptive processes of class I viruses. Because of the breadth of the subject and rapidity of new developments, it is inevitable that certain aspects of replication presented here may not be considered the latest word by every expert. To keep the article within manageable limits, little information has been presented on defective viruses and nothing on virus population genetics. It should be remembered, too, that an overview can include only a fraction of the detailed information on viral replication available in original articles and comprehensive reviews.
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The individual structural polypeptides of vesicular stomatitis virus have been examined by tryptic peptide analysis of 35S-methionine preparations labelled in vivo and 125I-preparations labelled in vitro. Isolates of the two classical serotypes of the virus (Indiana and New Jersey) and of a sub-type of the Indiana serotype, Brazil virus, were compared. The study showed that the major internal proteins of all three viruses gave similar maps, whereas the surface glycoproteins gave distinct maps that had very few spots in common. The map of the glycoprotein of Brazil virus, which has been shown previously to be more closely related serologically to Indiana virus than to New Jersey virus, did not show any greater similarity to the Indiana virus than to the New Jersey virus glycoprotein. On the other hand, peptide maps of the nucleoprotein and matrix protein showed Indiana and Brazil viruses to be more closely related to each other than to New Jersey virus.
Article
Vesicular stomatitis virus (VS) is an enveloped Rhabdovirus which is capable of productively infecting a multitude of hosts including many mammalian species, fish, and insects (Howatson, 1970; Bussereau, 1973). Two serotypes of VS virus have been identified: VSIndiana and VSNew Jersey (Pringle, 1975). The standard infectious virions of either serotype are rods (rhabdos = rod) of approximately 175 × 68 nm which have one hemispherical and one planar end and contain a helical nucleocapsid of approximately 30 equal turns followed by 4 or 5 diminishing coils at the rounded end (Howatson, 1970) (Fig. 1). Thus, virions appear bullet-shaped. Numerous “spikes” measuring 10 nm in length project externally from the entire surface of the viral envelope. Defective interfering (DI) particles which have the same morphology but are shorter and contain incomplete genomes are also produced at high multiplicities of infection. The viral envelope is acquired during maturation when nucleocapsids bud through pre-existing cell membranes; the site of budding varies according to the host species and can be on either the plasma membrane or the membranes of intracytoplasmic vesicles or on both (Zee et al., 1970). Fig. 1. Schematic representation of the vesicular stomatitis virion. Spikes (G protein) are shown attached to the planar end although there is no direct evidence that they are normally located there
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The ability of certain vesicular stomatitis virus (VSV; Indiana serotype) temperature-sensitive (ts) mutants to synthesize intracellular viral complementary RNA (vcRNA) at permissive or nonpermissive temperatures for productive infections has been investigated. Mutants belonging to complementation groups II, III, and V synthesize RNA at nonpermissive temperature in amounts essentially equivalent to that obtained at permissive temperatures. Mutant ts G I-114 possesses a thermolabile transcriptase and does not synthesize vcRNA at 40 degrees C; however, mutants ts O I-5, O I-53, O I-78, and O I-80 possess thermostabile transcriptases that are capable of some vcRNA synthesis at 40 degrees C. All five group I mutants are defective in their secondary transcription ability at 40 degrees C. Wild-type VSV New Jersey virus is able to complement the transcription defect of ts G I-114 at 40 degrees C. This complementation is inhibited by puromycin, suggesting that a viral gene product of VSV New Jersey (e.g., its transcriptase or a transcriptase component) is involved. Mokola virus is not able to complement the ts G I-114 defect, although Mokola does synthesize vcRNA in infected cells (in the presence or absence of cycloheximide).
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Inactivated defective interfering and complete particles of vesicular stomatitis virus given intracerebrally to adult mice protect them against challenge with homologous virus whether this is given at the same time or several days later. Two separate protective processes appear to be involved. The first, which comes into operation immediately after inoculation, is also effective against heterologous strains of vesicular stomatitis virus, rabies (another rhabdovirus), and a neurotropic strain of foot-and-mouth disease virus. The second, later effect, which is strain specific, appears to be correlated with the appearance of circulating neutralizing antibody. Our results suggest that the protective effect that Holland and his colleagues described using defective interfering particles of vesicular stomatitis virus may also be accounted for by an immunological mechanism rather than one involving interference.
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Three New Jersey serotype isolates of vesicular stomatitis virus (VSV) were found to produce a population of defective interfering (DI) particles heterogeneous in size and in their RNAs. Only Ogden and Glasgow isolates produced DI particles containing a well-defined major RNA component. Annealing of Ogden mRNAs to Glasgow virion RNA revealed that the two isolates had approximately 24% homologies in their nucleotide sequences. One short DI particle, generated by the Glasgow isolate, contained only approximately 230 nucleotides homologous to the Ogden virion RNA. In spite of this lack of homology, the particle fully interfered with infections by Ogden virions. A previously described Indiana serotype DI particle (HR), which was shown to interfere heterotypically with Prevec's New Jersey virion, also contained only approximately 260 nucleotides homologous to the RNA of this isolate. None of the New Jersey DI particles exhibited the ability to fully interfere heterotypically with Indiana serotype virion infections. DI particles, generated by a given viral isolate, contained nucleotide sequences complementary to a part of their 30 S mRNAs, and the sequences of shorter particle RNAs were always contained in the RNA of the larger particles. In this respect, they resembled the nonheterotypically interfering DI particles of the Indiana serotype.
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The relationships among the genomes of various rhabdoviruses belonging to the vesicular stomatitis virus subgroup were analyzed by an oligonucleotide fingerprinting technique. Of 10 vesicular stomatitis viruses, Indiana serotype (VSV Indiana), obtained from various sources, either no, few, or many differences were observed in the oligonucleotide fingerprints of the 42S RNA species extracted from standard B virions. Analyses of the oligonucleotides obtained from RNA extracted from three separate preparations of VSV Indiana defective T particles showed that their RNAs contain fewer oligonucleotides than the corresponding B particle RNA species. The fingerprints of RNA obtained from five VSV New Jersey serotype viruses were easily distinguished from those of the VSV Indiana isolates. Three of the VSV New Jersey RNA fingerprints were similar to each other but quite different from those of the other two viruses. The RNA fingerprints of two Chandipura virus isolates (one obtained from India and one from Nigeria) were also unique, whereas the fingerprint of Cocal virus RNA was unlike that of the serologically related VSV Indiana.
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Specific antisera were raised by immunization of rabbits with purified nucleocapsids containing only RNA and N protein (ribonucleoprotein, RNP) obtained from vesicular stomatitis (VS) virions of the Indiana (VSInd) and the New Jersey (VSNJ) serotypes. The specificity of anti-RNPInd serum was demonstrated by selective precipitation of homotypic RNPInd devoid of L and NS proteins; anti-RNPInd serum also selectively precipitated soluble N protein present in cytoplasm of infected cells, but co-precipitated a limited amount of contaminating soluble NS protein. Immunoglobulins prepared from each homotypic antiserum markedly inhibited in vitro transcription of VSInd and VSNJ virions. Anti-RNPInd and anti-RNPNJ immunoglobulins also exhibited cross-reactivity by inhibiting transcription of heterotypic virions, but only to a much lesser degree than in the homotypic reaction. Anti-RNPInd immunoglobulin did not inhibit transcription of the antigenically unrelated Chandipura rhabdovirus, but anti-RNPNJ immunoglobulin did to a very limited extent. The transcription inhibitory activity of anti-RNPInd immunoglobulin was not dependent on RNP immunoprecipitation activity, which could be diluted out well before loss of antitranscriptase activity. Anti-RNPind immunoglobulin appeared to exert its effect on transcription by blocking elongation rather than initiation or reinitiation of RNA transcripts.
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Vesicular stomatitis New Jersey (VS NJ) virus is capable of undergoing rapid evolution in nature and therefore has the potential for antigenic variation. We selected an area of Costa Rica where VS NJ virus is enzootic to study whether this virus used the mechanism of antigenic variation to persist in nature. Three sentinel herds and three nonsentinel herds were observed from 1986 to 1988. Eleven VS NJ virus isolates were collected from naturally infected cattle. Remarkably, nine animals that were bled prior to reinfection with VS NJ virus had neutralizing antibody titers up to 1: 102,400 yet virus was isolated from, and disease was observed in, these animals. Sequence analysis of the portion of the glycoprotein gene coding for the neutralizing epitopes demonstrated that all virus isolates were 98-100% similar with no indication of specific genetic variation. The 3' end of the glycoprotein gene also remained stable in that all isolates were again 98-100% similar in nucleotide sequence. Each isolate was neutralized to equivalent titers with monoclonal antibodies directed against four neutralizing epitopes on the glycoprotein. Additionally, preisolation sera from each animal were able to neutralize the virus that caused the subsequent disease. These results provide evidence that antigenic variation is not a mechanism used by VS NJ virus to persist in an enzootic focus of Costa Rica.
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The nucleotide sequences at the 5' and 3' termini of RNA isolated from the New Jersey serotype of vesicular stomatitis virus [vsV(NJ)] and two of its defective interfering (DI) particles have been determined. The sequence differs from that previously demonstrated for the RNA from the Indiana serotype of VSV at only 1 of the first 17 positions from the 3' terminus and at only 2 of the first 17 positions from the 5' terminus. The 5'-terminal sequence of VSV(NJ) RNA is the complement of the 3'-terminal sequence, and duplexes which are 20 bases long and contain the 3' and 5' termini have been isolated from this RNA. The RNAs isolated from DI particles of VSV(NJ) have the same base sequences as do the RNAs from the parental virus. These results are in sharp contrast to those obtained with the Indiana serotype of VSV and its DI particles, in which the 3'-terminal sequences differ in 3 positions within the first 17. However, with both serotypes, the 3'-terminal sequence of the DI RNA is the complement of the 5'-terminal sequence of the RNA from the infectious virus. These findings suggest that the 3' and 5' RNA termini are highly conserved in both serotypes and that the 3' terminus of DI RNA is ultimately derived by copying the 5' end of the VSV genome, as recently proposed (D. Kolakofsky, M. Leppert, and L. Kort, in B. W. J. Mahy and R. D. Barry, ed., Negative-Strand Virus and the Host Cell, 1977; M. Leppert, L. Kort, and D. Kolakofsky, Cell 12:539-552, 1977; A. S. Huang, Bacteriol. Rev. 41:811-8218 1977).
Article
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The in vitro RNA synthesis by the virion-associated RNA polymerase of vesicular stomatitis virus (VSV), New Jersey serotype, was compared with that of the serologically distinct Indiana serotype of VSV. The New Jersey serotype of VSV synthesized five distinct mRNA species in vitro, three of which were smaller than the corresponding species synthesized by the Indiana serotype of VSV. These included the mRNA's coding for the G, M, and NS proteins. By hybridization experiments, virtually no sequence homology was detected between the mRNA's of the two serotypes. Despite this lack of overall homology, the 12 to 18S mRNA species of both serotype contained a common 5'-terminal hexanucleotide sequence, G(5')ppp(5')A-A-C-A-G. The signicance of this finding in light of specific interactions between the two serotypes of VSV in vivo is discussed.
Article
Rescue virions obtained after superinfection of ts O45(V) Indiana serotype-infected cells by uv-irradiated vesicular stomatitis virus (VSV) at a nonpermissive temperature do not incorporate G protein synthesized at this temperature. They apparently contain G protein in their envelope since they are neutralized by homotypic antivirion and antispike sera. Rescue of ts O45(V) mutant Indiana serotype by uv-irradiated VSV New Jersey serotype is demonstrated. Ultraviolet-inactivated VSV of New Jersey serotype has therefore been used to determine the origin of the incorporated G protein molecules. The progeny-rescue virions belong genetically to VSV Indiana complementation group V. They are however neutralized by anti-New Jersey serum and not by anti-Indiana serum. Rescue ts O45(V) virions have thus probably reincorporated G protein molecules supplied by the uv-irradiated New Jersey virus. This provides further evidence suggesting that protein G is encoded for by gene V.
Article
The methionme-containing tryptic peptides of the five constituent polypeptides of members of the vesicular stomatitis groups of rhabdoviruses were compared to the Indiana serotype by two-dimensional thin-layer chromatography-electrophoresis. While the corresponding protein of each virus serotype could be identified by its characteristic map, there were apparent common peptides shared between serologically related members. Three methionine-containing peptides of the N protein of VSV (Indiana) could not be separated from peptides of VSV (Cocal) and of these three, one peptide was also present in VSV (New Jersey). A comparison of the tryptic peptides of proteins from two members of the VSV (New Jersey) serotype revealed characteristic differences in the polypeptides from the closely related, but serologically distinguishable, New Jersey-Concan and New Jersey Missouri isolates.
Article
The partial cleavage products resulting from treatment of [35S]methionine-labeled vesiculovirus proteins with N-chlorosuccinimide were analyzed by SDS-polyacrylamide gel electrophoresis. The resultant cleavage patterns of the nucleocapsid (N) and matrix (M) proteins of five serologically distinct members of this group, while unique, showed a number of similarities in fragment pattern and position. The relative location of tryptophan in the N proteins shows considerable similarity between Indiana and Cocal serotype and between Chandipura and Piry serotypes. On the other hand, the M protein pattern demonstrates similarities between Indiana, Cocal, New Jersey, and Piry serotypes but is quite distinct for the Chandipura serotype. The cleavage pattern of N and M proteins of representative viruses of the two New Jersey subtypes showed that their tryptophan positions are essentially identical. The potential usefulness of this mapping procedure is further considered.
Article
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The effects of trypsin and chymotrypsin on the infectivity, morphology and antigenic properties of the Indiana and Brazil strains of vesicular stomatitis virus have been studied. Each enzyme reduced the infectivity of the Indiana strain by about 10,000 fold but the infectivity of the Brazil strain was unaffected. Electron microscopy of the treated particles and polyacrylamide gel electrophoresis of the viral polypeptides showed that each enzyme removed all the surface projections of the Indiana virus but about one third of the projections of the Brazil virus were resistant. Complement fixation tests showed that, in contrast to the complete removal of the antigenic activity from the surface of the Indiana virus, about one third of the surface antigenic activity of the Brazil strain was retained. The enzyme resistant projections of the Brazil virus absorbed neutralizing antibody from the homologous antiserum but were much less active in stimulating the production of neutralizing antibody in guinea pigs although the level of complement fixing antibody produced was similar to that produced by the intact virus. Amino acid analysis also gave differences between the total and enzyme resistant surface projections. These results suggest that, in contrast to the Indiana virus, the Brazil virus possesses two surface projections which differ in their resistance to proteolytic enzymes.
Article
The structural proteins of several isolates of the Indiana serotype (Orsay, Glasgow, and San Juan) of vesicular stomatitis virus (VSV) were compared by sizing peptides generated by digestion with Staphylococcus aureus V8 protease or α-chymotrypsin. These digestions yielded nearly identical peptide patterns for all the VSV proteins except for the glycoprotein, indicating that the major surface antigen, even of these closely related isolates, is subject to change. When two serologically distinct VSV isolates (Indiana and New Jersey) were similarly examined, each protein had a peptide pattern characteristic of that serotype. The glycoprotein of the two serotypes had no peptides in common, while the internal viral proteins L, N, NS, and M showed various degrees of similarity. Digests of the L (polymerase) and M (membrane) protein, in particular, generated groups of small peptides with identical mobilities, suggesting that there may be core regions in the Indiana and New Jersey strain which are conserved. These conserved regions may in turn represent essential functions of rhabdovirus proteins and are promising targets for broad-spectrum antivirals.
Article
The rescue at nonpermissive temperature of complementation group II temperature-sensitive mutants of vesicular stomatitis virus (VSV) by uv-irradiated virus is analyzed. Rescue is group-characteristic. The helper virus can be either the wt strain or a mutant belonging to any group of ts mutants except group II. With regard to genotype, the rescue progeny virus is temperature-sensitive and belongs to group II, and its ts II parent [ts O52(II)] can be characterized. As for phenotype, the in vitro thermal stability of rescue virions is intermediate between that of parental ts O52(II) and uv-irradiated wt virus suggesting incorporation of some wt protein II molecules in the rescue virions. These molecules are: first, the transcription products of the surviving genes II+ of the irradiated virus and second, the structural molecules of protein II+ supplied by the irradiated virus. This nongenetic complementation is demonstrated by action spectra analysis [Deutsch, V., Muel, B., and Brun, G. (1977). Virology77, 294–305], rescue in the absence of simultaneous protein synthesis, generally observed relationships between uv dose and effect which can be represented by fitted straight lines with a shallow slope accounting for the inactivation of a protein target, efficiency of rescue at uv doses high enough to inactivate every gene II+, rescue by irradiated ts G114(I) mutant virions and by irradiated defective-interfering short T particles, and multiplicity dependence contrasting with that evidenced when rescue could be mainly ascribed to gene survival. Different slopes (zero or different from zero) were seen in dose-effect curves representing rescue obtained by structural protein molecules, suggesting that protein II structural role could be distinguished from its functional role(s) by uv sensitivity. Differences in efficiency of the rescue of ts O52(II) by ts I mutants irradiated with low uv fluence may reflect their different transcribing capabilities at 39.6°. The results are discussed taking into account the fact that the phenotype of group II mutants is characterized by an unstable nucleocapsid.
Article
Noninfectious spikeless particles have been obtained from vesicular stomatitis virus (VSV, Indiana serotype) by bromelain or Pronase treatment. They lack the viral glycoprotein (G) but contain all the other viral components (RNA, lipid, and other structural proteins). Triton-solubilized VSV-Indiana glycoprotein preparations, containing the viral G protein as well as lipids (including phospholipids), have been extracted from whole virus preparations, freed from the majority of the detergent, and used to restore infectivity to spikeless VSV. The infectivity of such particles has been found to be enhanced by poly-L-ornithine but inhibited by Trition or homologous antiserum pretreatment. Heat-denatured glycoprotein preparations were not effective in restoring the infectivity to spikeless VSV. Heterologous glycoprotein preparations from the serologically distinct VSV-New Jersey serotype were equally capable of making infectious entities with VSV-Indiana spikeless particles, and the infectivity of these structures was inhibited by VSV-New Jersey antiserum but not by VSV-Indiana antiserum. Purified, detergent-free glycoprotein selectively solubilized from VSV-Indiana by the dialyzable detergent, octylglucoside, also restored infectivity of spikeless virions of VSV-Indiana and VSV-New Jersey.
Article
Recently accumulated knowledge allows more precise comparison of the structural (and possibly evolutionary) relationships of several different animal rhabdoviruses: vesicular stomatitis virus, rabies virus, Kern Canyon virus, and spring viremia of carp virus. Each virus is composed primarily of a glycoprotein, an RNA-associated nucleoprotein, and one or two membrane proteins. Vesicular stomatitis virus group viruses contain lesser amounts of two additional distinct polypeptides, NS and L. The separate viruses undergo structural polypeptide phosphorylation in vivo according to characteristic patterns. In vesicular stomatitis virus the NS protein is selectively phosphorylated. In rabies group viruses and in spring viremia of carp virus, the nucleoprotein is the predominant phosphoprotein; in these viruses only the phosphorylated moiety is selectively cleaved off with trypsin. In Kern Canyon virus, only membrane protein and glycoprotein are weakly phosphorylated. Each virus possesses a virion-bound protein kinase. Vesicular stomatitis virus group viruses, Kern Canyon virus, and spring viremia of carp virus only contain virion-bound transcriptases of respectively decreasing levels of activity demonstrable in vitro. Vesicular stomatitis and Kern Canyon viruses replicate efficiently in enucleated cells; rabies virus does not. Based upon these observations, it is suggested that vesicular stomatitis virus may represent the most highly evolved of these rhabdoviruses, whereas spring viremia of carp and Kern Canyon viruses may represent "evolutionary links" between the vesicular stomatitis and rabies virus groups.
Article
This experiment assess different MI for Vesicular Stomatitis VS virus industrial culture production and evaluated the effect of glycoprotein G concentration in relation to antibodies induction against VS on guinea pigs vaccinated with oil bivalent vaccine (Indiana I and New Jersey NJ). With efficient MI it was possible to get better kinetic of infection at industrial cultures, reducing time of culture and improving viral titers. In addition, it was found that neutralizing titers of guinea pigs immunized with an EV vaccine containing 5 micrograms of glycoprotein G, were 3.66 log10 for I and 4.06 log10 for NJ, which are correlated to protection titers in cattle. About this study can be concluded that selecting a superior MI, efficiency of industrial VE virus production can be improved; on the other hand, glycoprotein G quantification methodology can be useful for a good quality VS Vaccine industrial manufacture.
Chapter
Vesicular stomatitis virus (VSV) belongs to the family of rhabdoviruses. This group of viruses includes VSV, rabies virus, Marburg virus, and several other viruses that replicate in anthropoids as well as in other mammals.(1,2) The natural hosts of these viruses are listed in Table I. One of the major mor phologic features of this group of viruses is their bullet-shaped virion. These bullet-shaped rods are cylinders approximately 170 nm in length and 70 nm in diameter. Each particle consists of a nucleoprotein (RNA) core helically wound around a central axial hollow. This is enclosed in an envelope consisting of a lipid bilayer and associated structural proteins. In the VSV group, there are distinct strains, including Indiana, New Jersey, Brazil, Argentina, Cocal, Piry, and Chandipura. All the strains except Piry and Chandipura share some common antigens; those two strains appear to be totally unrelated antigenically to others.(3) According to the nomenclature of Wagner et al.,(4) proteins of the Indiana serotype are designated L, G, N, NS, and M and have molecular weights of 190, 69, 50, 45, and 29 kDa, respectively.(5)
Chapter
The various categories of infectious agents that confront higher vertebrate hosts are quite different with respect to acuteness and rapidity of the disease process they incite. Consequently, the evolutionary or selective pressure that ultimately determines whether the relationship between parasite and host becomes symbiotic or parasitic is also decisive in shaping a particular part of the host physiology specifically for that function. Since most parasites have an enormously faster generation cycle than the vertebrate host, the parasites’ adaptive capacity is much the greater of the two. The ultimate equilibrium of the host-parasite relationship and their relative capacity to adapt mutually are obviously determined by both parties. What establishes this balance is poorly understood, but, viewed teleologically, survival of both parasite and host seems to keep the relationship in check. Within the vertebrate host whose reactive systems are called into play by such infections, immune responses by antibodies and cells (reviewed in Notkins, 1975, Bloom and Rager-Zisman, 1975; Blanden et al., 1976b), nonspecific resistance by macrophages (Lindenmann et al., 1978; Cheers and McKenzie, 1978) or other cells, past and present experience with infection, as well as concurrent infections all play some part.
Chapter
Chapter 1 describes the general features of the family Rhabdoviridae, and Chapter 3 provides a moderately detailed analysis of the rhabdovirus membrane. This chapter is designed to set the stage for the in-depth chapters that follow by providing a general description of the structural components, infectivity, host reactivity, and immunology of rhabdoviruses. As is true of all but Chapters 1 and 9–11, this chapter is concerned primarily with the best-studied and prototypic rhabdovirus, vesicular stomatitis virus (VSV). It seems reasonable to assume that the structural components that comprise the VSV virion are similar to those of other rhabdoviruses, but comparisons will be made, where appropriate, with the rabies subgroup and rhabdoviruses of other animals. It should be stated from the outset that productive infection by VSV of mammalian and avian cells far exceeds that of rabies and other animal rhabdoviruses, such as those of fish. Nothing will be said in this chapter about the structure and infectivity of the plant rhabdoviruses, which are discussed in Chapter 10.
Article
Following the discovery of the androgens in the early nineteen thirties and the first demonstration of their anabolic effects in 1935, evidence rapidly accumulated to show that practically no tissue in the body was immune from possible androgen stimulation, whether assessed on histological or on biochemical criteria. Considering the diversity of the biochemical effects attributable to the androgens, it is not surprising that an equal diversity is to be seen in the experiments designed to study these effects. Arbitrarily, although with some justification on historical and biochemical grounds, these studies can be said to fall into two main groups.
Chapter
This chapter describes the biological characteristics of two viruses coming from two very different hosts. The starting point was the finding of sigma virus of Drosophila and vesicular stomatitis virus of mammals are very similar morphologically. Such comparisons are not frequently made, but may be of interest as contributions to a phylogenetic classification of viruses. Such researches may also be useful in relating poorly known viruses to well described ones. Sigma virus is a hereditary infectious agent relatively frequently present in natural populations of Drosophila. It confers on infected flies, a peculiar symptom, called carbon dioxide (CO2) sensitivity. VS virus is the causative agent of a rather mild disease of horses and cattle, which is widespread in the New World. It has been extensively investigated in many laboratories and is accepted as the prototype virus of the Rhabdovirus group. Two main serotypes exist, called Indiana and New Jersey. A recently observed third strain, the Coccal virus, is serologically related to the Indiana serotype. In the work described in this chapter, the Indiana serotype of VS virus is used. Recently, studies with other serotypes have been made.
Article
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Vesicular stomatitis (VS) is a viral disease of horses, cattle and swine caused by vesiculoviruses (VSV) of the Rhabdoviridae family. The disease is a zoonosis. VS is endemic in the Western Hemisphere, where its incidence is confined to the warmer regions of North America, as well as Central and South America with tropical climate. The article presents the most important issues concerning the historical background and current status of the disease and its etiological agent. The impact of climatic and ecological conditions on the emergence of new outbreaks and virus strain evolution is discussed, as well as the role of insects in the epidemiology of infections and limited possibilities of specific prevention. The importance of VS virus as a vaccine and oncolytic vector and prospects for its use in the prevention of human infectious diseases and cancer therapy are also highlighted.
Chapter
Vesicular stomatitis viruses (VSVs) and rabies virus are members of the family Rhabdoviridae. Vesicular stomatitis (VS) in cattle and pigs resembles foot‐and‐mouth disease and in pigs swine vesicular disease or Seneca Valley virus. The presence of VS in livestock results in export restrictions on livestock from VS‐affected to VS‐free zones. When vesicular disease is observed in swine, steps should be taken to stop the movement of animals and materials onto or off the premises until a diagnosis is made. The appropriate animal health authorities should be notified immediately. Preventive measures include insect control, housing animals indoors during peak insect feeding times, and implementation of biosecurity procedures designed to avoid the introduction of VSV from affected premises via equipment, personnel, or animals. The best way to prevent rabies is to control the disease in wildlife reservoirs and use physical barriers to protect swine from wildlife.
Article
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There are two major serotypes of vesicular stomatitis virus (VSV), Indiana (VSIV) and New Jersey (VSNJV). We recovered recombinant VSIVs from engineered cDNAs that contained either (i) one copy of the VSIV G gene (VSIV-GI); (ii) two copies of the G gene, one from each serotype (VSIV-GNJGI); or (iii) a single copy of the GNJ gene instead of the GI gene (VSIV-GNJ). The recombinant viruses expressed the appropriate glycoproteins, incorporated them into virions, and were neutralized by antibodies specific for VSIV (VSIV-GI), VSNJV (VSIV-GNJ), or both (VSIV-GNJGI), according to the glycoprotein(s) they expressed. All recombinant viruses grew to similar titers in cell culture. In mice, VSIV-GNJ and VSIV-GNJGI were attenuated. However, in swine, a natural host for VSV, the GNJ glycoprotein-containing viruses caused more severe lesions and replicated to higher titers than the parental virus, VSIV-GI. These observations implicate the glycoprotein as a determinant of VSV virulence in a natural host and emphasize the differences in VSV pathogenesis between mice and swine.
Article
This chapter focuses on the important discovery that virus-specific cytotoxic T cells are dually specific for virus and for a self cell surface antigen encoded by the major histocompatibility complex (MHC). The initial work was carried out on the lymphocytic choriomeningitis virus system but it soon became evident that the same phenomenon applied to many other viruses. In addition, the same principle has been found to hold for other antigenic systems, such as trinitrophenyl coupled to cells, minor histocompatibility antigens, and the H-Y model. Graft rejection and the need for genetically homogeneous inbred mouse strains for cancer research led to the development of transplantation immunology and immunogenetics. The result is that the gene complex coding for major transplantation antigens is one of the better understood mammalian genetic regions. Cytotoxic T-cell specificity is comparable to serological specificity. Because quantification of specificity or cross-reactivity is difficult, and because of the technical limitations of these cytotoxic T-cell assays, results are interpreted with great reservation. MHC restriction reflects the fact that the effector function of T cells is determined by the kind of Self-H recognized together with the foreign antigen on cell surfaces: K and D are receptors for lytic signals, I determinants are receptors for cell differentiation signals that are delivered antigen-specifically by T cells.
Article
Monoclonal antibodies reactive with the major surface glycoprotein (G-protein) of vesicular stomatitis virus serotypes Indiana and New Jersey (VSV-Ind, VSV-NJ) have been isolated and characterized. The reactivity of each monoclonal was determined by enzyme-linked immunosorbent assay (ELISA), competitive binding assay (CBA), and the ability to neutralize infectivity. It was found that the majority of the antibodies were of the IgG2a subclass. In the CBA, unlabeled monoclonal antibodies were used to compete for radiolabeled antibodies in binding to solid-phase immunoadsorbents. The VSV-NJ G-protein appears to contain four nonoverlapping epitopes by these analyses. However, the VSV-Ind G-protein is more complex since four epitopes were defined which exhibited varying degrees of overlap. In some cases, this overlap was defined by complete reciprocal competition between antibodies with different reactivity patterns. In other instances, partial or nonreciprocal competition between antibodies was observed. These results may indicate epitopes in close proximity or suggest allosteric modifications in the G-protein induced by antibody binding. A fifth epitope on the Ind G-protein was defined by a monoclonal antibody which could bind to the G-proteins of both VSV-Ind and VSV-NJ but could only neutralize infectivity of the VSV-Ind serotype.
Thesis
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Foot-and-mouth disease (FMD) is the most contagious disease of mammals and has great potential to cause severe economic losses in split-hoofed animals. In disease management, decisions often must be made in a context of uncertainty. However, epidemiological models can be a powerful tool to aid the development of animal health policies and preventive disease control. The work aimed to find priority regions for vesicular disease surveillance in Brazil. The work was conducted using data from the Continental Information and Surveillance System (SivCont) for Brazil. The data refer to Vesicular Syndromes, collecting information on FMD and Vesicular Stomatitis (VS), as well as other diseases with symptoms compatible with vesicular syndromes of diverse infectious and noninfectious sources. It is also essential to consider the timeliness involved in surveillance programs, which can demonstrate the dynamics and interaction of the activities carried out. Also, the risk-based requirements of the FMD surveillance system were assessed. For the different analyses, various techniques were implemented, such as GLM, data mining, and Bayesian network analysis. On the other hand, the results of chapter 3 revealed delays in the timeliness of each state, the different classifications of diseases and especially in conditions with symptoms equal to FMD. Also, it even showed that there is a significant variation in the timeliness of the FMD surveillance system when the Brazilian states were compared. It was also observed in chapter 4 that diseases with symptoms like FMD are the most notified events, and occur with more frequency in SC and PR. Besides this, the states of MG, PA, MS, RO, and GO are identified as being more likely to have positive FMD diagnosis and more delayed in notifying. All results obtained in this research will allow decision-makers in the official veterinary services to strengthen surveillance measures in states with extreme timeliness values. Thus, reinforces the FMD surveillance system, which supports the surveillance programs. Keywords: Foot-and-Mouth disease, risk-based surveillance, prioritization, GLM, Bayesian network
Article
Nineteen independently isolated hybridomas producing monoclonal antibodies to the glycoprotein of vesicular stomatitis virus were isolated and studied for their capacity to neutralize viral infectivity. By measuring competitive binding of 125I-labeled monoclonal antibodies in a radioimmunoassay. 11 different, non-cross-reacting antigenic determinants were identified on the vesicular stomatitis virus G protein. All monoclonal antibodies reacting with determinants 1, 2, 3, and 4 resulted in viral neutralization, whereas those binding to the other seven determinants did not neutralize infectivity. The mixture of two monoclonal antibodies binding to different determinants resulted in a more rapid neutralization than either antibody alone, suggesting that different antibodies can exert a synergistic effect on viral neutralization. Kinetic experiments revealed biphasic neutralization curves similar to those expected for heterologous antibody. No evidence could be obtained to relate biphasic kinetics of viral neutralization to heterogeneous populations either of antibody molecules or of virus. The possible significance of the kinetic data with monoclonal antibodies is discussed.
Chapter
Cell culture is an integral part of virology as the viruses are the obligate intracellular parasites that require replication inside a living cell to produce copies of themselves. Since the introduction of cell culture, a number of cell culture systems have been developed which are in use to isolate, propagate, and study growth kinetics vis-à-vis host–virus interactions of a number of virus species affecting diverse range of hosts. These systems comprise of primary cell culture and cell lines having finite or infinite life span. Owing to their immortality and other advantages, continuous cell lines are the most frequently used category of cell culture system for animal viruses. During recent years, traditional cell culture system has changed from the use of glass bottles/flasks to the use of plastic tissue culture flasks and diverse range of culture media have been developed to meet the research needs. This chapter elaborates various cell culture systems which have been developed and are most frequently used for isolation and propagation of common DNA and RNA viruses affecting pigs.Key wordsPrimary cell cultureCell linesSecondary cell culturePorcine viruses
Article
In search of an anti-transcriptase, antibody was raised in rabbits to partially purified, soluble NS protein present in cytoplasmic extracts of cells infected with the Indiana serotype of vesicular stomatitis (VSInd) virus. This antiserum gave specific reactions of identity by agar immunodiffusion with both cytoplasmic and virion NS protein. NS antiserum also preferentially precipitated NS 3-H-labeled protein from infected cytoplasmic extracts, whereas anti-whole VSInd virion serum also precipitated N 3-H-labeled protein from extracts both of infected cytoplasm and virion nucleocapsids. Transcriptase activity of VSInd cytoplasmic or virion-derived nucleocapsids was effectively inhibited by ribonuclease-free immunoglubulin prepared from homologous NSInd antiserum or from anti-whole vesicular stomatitis virus serum. Transcriptase activity of heterologous New Jersey serotype (VSNJ) nucleocapsids and virions was not appreciably affected by anti-NSInd or by anti-whole VSInd virion gamma globulin. Anti-NS gamma glubulin immediately switched off RNA synthesis by actively transcribing VSInd nucleocapsids, a finding which suggests that NS antibody inhibits RNA chain elongation.
Article
We propose a reclassification of five strains of the New Jersey serotype of vesicular stomatitis virus into two subtypes designated Concan and Hazelhurst. This subclassification into two subtypes is based on reciprocal differences in antibody neutralization of virion infectivity, nucleotide base sequence homology, oligonucleotide maps of virion RNA, and interference by defective-interfering particles.
Article
We report the entire glycoprotein (G) gene nucleotide sequences of 26 vesicular stomatitis virus Indiana serotype (VSV IND) type 1 isolates from North and Central America. These sequences are also compared with partial G gene sequences of VSV IND type 2 (Cocal) and type 3 (Alagoas) viruses and the complete G gene sequences of the more distantly related VSV New Jersey (NJ) and Chandipura viruses. Phylogenetic analysis of the G gene sequences by maximum parsimony revealed four major lineages or subtypes within the classical VSV IND (type 1) viruses, each with a distinct geographic distribution. A high degree of VSV genetic diversity was found in Central America, with several virus subtypes of both VSV IND and NJ serotypes existing in this mainly enzootic disease region. Nineteen percent sequence variation but no deletions or insertions were evident within the 5' noncoding and the coding regions of the VSV IND type 1 G genes. In addition to numerous base substitutions, the 3' noncoding regions of these viruses also contained numerous base insertions and deletions. This resulted in striking variation in G gene sizes, with gene lengths ranging from 1,652 to 1,868 nucleotides. As the VSV IND type 1 subtypes have diverged from the common ancestor with the NJ subtypes, their G mRNAs have accumulated more 3' noncoding sequence inserts, ranging up to 303 nucleotides in length. These primarily consist of an imprecise reiteration of the sequence UUUUUAA, apparently generated by a unique polymerase stuttering error. Analysis of the deduced amino acid sequence differences among VSV IND type 1 viruses revealed numerous substitutions within defined antigenic epitopes, suggesting that immune selection may play a role in the evolution of these viruses.
Article
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The antigens of the nucleoprotein core and the coat of vesicular stomatitis virus (VSV) particles of the Indiana serotype were prepared and purified by sucrose gradient fractionation. Antibody was prepared separately to each of the two antigen fractions. By immunological procedures, it was shown that soluble antigens of VSV preparations sedimenting at 20S and in the leading edge of the 6S region are antigenically related to VP3, the protein of the virus core, whereas the 6S soluble antigen cross-reacts only with viral coat antibodies. These results confirm previous results obtained by polyacrylamide gel analysis of the antigens. It has further been demonstrated that the 6S antigen is a glycoprotein. Comparing antigens of the New Jersey and Indiana serotype showed that the coat antigens of virus particles and the 6S antigen are immunologically distinct in the two serotypes. In complement-fixation tests, the core antigens and the soluble 20S antigens from one serotype showed a cross-reaction with antiserum prepared against core proteins of the other serotype.
Article
SUMMARY Treatment of the infective component of vesicular stomatitis virus with Nonidet P4o produces an infective skeleton-like structure which has the shape and approxi- mate size of the intact virus particle. The infectivity of the skeleton is enhanced I oo to I ooo fold by mixing with DEAE-dextran. The skeleton lacks the outer envelope and fringe structure and in consequence does not produce neutralizing antibodies in guinea pigs. The density of the skeleton is 1.22 g./ml, in potassium tartrate gradients compared with x "t4 g./ml, for the virus. Sodium deoxycholate removes protein from the skeleton and releases the filamentous ribonucleoprotein in an infective form. As with the skeleton, the infectivity of the ribonucleoprotein is enhanced by DEAE- dextran. Ribonuclease has no effect on the ribonucleoprotein but trypsin destroys its infectivity. The ribonucleoprotein has a density of ~-22 g./ml, in tartrate gradients, sediments at about ~40 s in sucrose gradients and does not produce neutralizing antibodies in guinea-pigs.
Article
Our experiments on the stepwise degradation of vesicular stomatitis virus into well characterized structural components have enabled us to relate some of the biological properties of the virus to certain of its structural units (Cartwright, Smale & Brown, 1969, 1970). For example, incubating the virus with trypsin produces a bullet-shaped structure similar to the virus except that it no longer possesses the surface projections. Unlike the virus, the projection-free component has low infectivity and does not produce neutralizing antibodies when inoculated into guinea-pigs, thus providing decisive evidence that the immunizing activity of the virus is associated with the surface projections. By treating the virus with Tween + ether or Nonidet P40 or sodium deoxycholate (Brown, Cartwright & Smale, 1967; Cartwright et al. 1970), the immunizing antigen can be released in a biologically active form, sedimenting at about 6s. An infective skeleton-like structure with the same size and shape as the virus is also produced by treating the virus with Tween + ether or with Nonidet.
Article
Sera collected from horses, swine, and cattle before inoculation with either the New Jersey or Indiana types of vesicular stomatitis virus and at periodic intervals thereafter, were tested by complement-fixation methods with antigens prepared from chick embryo tissues infected with these respective viruses. Convalescent sera from the three species differed in their behaviour in these tests. In horses as in guinea pigs, complement-fixing activity with homologous VS antigen appeared in about six to eight days after infection, reached a maximum titer in ten days to two weeks, remained relatively constant for seven to ten days then began to decline. No cross fixation between types was recorded. None of the cattle sera at any time exhibited any fixation in direct complement-fixation tests with homologous antigen. To demonstrate non-complement-fixing antibody in these convalescent bovine sera it was necessary to employ an indirect complement-fixation method using hyperimmune guinea pig or convalescent horse serum to detect residual antigen. The time of antibody development as indicated by the appearance of inhibitory activity in these indirect tests coincided with that demonstrated in horses by direct complement-fixing tests. The convalescent pig sera showed marked prozones effects (increased hemolysis) in lower serum dilutions. This increased hemolysis was traceable to certain substances present in normal as well as in convalescent swine serum and not to non-complement-fixing antibodies corresponding to those in convalescent bovine sera. Natural hemolysins for sheep red cells, third component of complement, and possibly other constituents, notably lipids, apparently contribute to this augmented hemolysis or reduced fixation of complement.
Workshop on ' Bridging Groups of Viruses Immunodiffusion studies of the antigenic relationships within and between serotypes of vesicular stomatitis virus The resin-agglutination test for vesicular stomatitis
  • F A Murpiqy
  • R E Shope
  • I Myers
  • W L Hanson
Proceedings of the Electron Microscopic Society of America, 27th meeting, p. 372. MURPIqY, F. A. & SHOPE, R. E. 097I)- Proceedings of the Second International Congress for Virology, Budapest. Workshop on ' Bridging Groups of Viruses', p. 26 I. MYERS, W.L. & HANSON, R. 1'. (t962). Immunodiffusion studies of the antigenic relationships within and between serotypes of vesicular stomatitis virus. American Journal of Veterinary Research 23, 896. ~ETO, F.M. & StoRE, D. 0958). The resin-agglutination test for vesicular stomatitis. American Journal of Veterinary Research XD, 761
The infection of cells by bullet-shaped viruses
  • R A Harrison
  • A K Whitfield
MURPHY, r. A., HARRISON, A. K. & WHITFIELD, S. G. 0969). The infection of cells by bullet-shaped viruses. Proceedings of the Electron Microscopic Society of America, 27th meeting, p. 372.
Workshop on ' Bridging Groups of Viruses
  • F A Murpiqy
  • R E Shope
MURPIqY, F. A. & SHOPE, R. E. 097I)-Proceedings of the Second International Congress for Virology, Budapest. Workshop on ' Bridging Groups of Viruses', p. 26 I.
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