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A new grass carp reovirus (GCRV), healthy grass carp reovirus (HGCRV), was isolated from grass carp in 2019. Its complete genome sequence was determined and contained 11 dsRNAs with a total size of 23,688 bp and 57.2 mol% G+C content, encoding 12 proteins. All segments had conserved 5' and 3' termini. Sequence comparisons showed that HGCRV was closely related to GCRV-873 (GCRV-I; 69.57–96.71% protein sequence identity) but shared only 22.65–45.85% and 23.37–43.39% identities with GCRV-HZ08 and Hubei grass carp disease reovirus (HGDRV), respectively. RNA-dependent RNA-polymerase (RdRp) protein-based phylogenetic analysis showed that HGCRV clustered with Aquareovirus-C (AqRV-C) prior to joining a branch common with other aquareoviruses. Further analysis using VP6 amino acid sequences from Chinese GCRV strains showed that HGCRV was in the same evolutionary cluster as GCRV-I. Thus, HGCRV could be a new GCRV isolate of GCRV-I but is distantly related to other known GCRVs. Grass carp infected with HGCRV did not exhibit signs of hemorrhage. Interestingly, the isolate induced a typical cytopathic effect in fish cell lines, such as infected cell shrank, apoptosis, and plague-like syncytia. Further analysis showed that HGCRV could proliferate in grass carp liver (L28824), gibel carp brain (GiCB), and other fish cell lines, reaching a titer of up to 7.5 × 104 copies/μL.
Ke Zhang; Wenzhi Liu; Yiqun Li; Yong Zhou; Yan Meng; Lingbing Zeng; Vikram Vakharia; Yuding Fan. Isolation, Identification, and Genomic Analysis of a Novel Reovirus from Healthy Grass Carp and Its Dynamic Proliferation In Vitro and In Vivo. Viruses 2021, 13, 690 .
AMA StyleKe Zhang, Wenzhi Liu, Yiqun Li, Yong Zhou, Yan Meng, Lingbing Zeng, Vikram Vakharia, Yuding Fan. Isolation, Identification, and Genomic Analysis of a Novel Reovirus from Healthy Grass Carp and Its Dynamic Proliferation In Vitro and In Vivo. Viruses. 2021; 13 (4):690.
Chicago/Turabian StyleKe Zhang; Wenzhi Liu; Yiqun Li; Yong Zhou; Yan Meng; Lingbing Zeng; Vikram Vakharia; Yuding Fan. 2021. "Isolation, Identification, and Genomic Analysis of a Novel Reovirus from Healthy Grass Carp and Its Dynamic Proliferation In Vitro and In Vivo." Viruses 13, no. 4: 690.
The grass carp reovirus (GCRV) causes severe hemorrhagic disease with high mortality and leads to serious economic losses in the grass carp (Ctenopharyngodon idella) industry in China. Oral vaccine has been proven to be an effective method to provide protection against fish viruses. In this study, a recombinant baculovirus BmNPV-VP35-VP4 was generated to express VP35 and VP4 proteins from GCRV type Ⅱ via Bac-to-Bac baculovirus expression system. The expression of recombinant VP35-VP4 protein (rVP35-VP4) in Bombyx mori embryo cells (BmE) and silkworm pupae was confirmed by Western blotting and immunofluorescence assay (IFA) after infection with BmNPV-VP35-VP4. To vaccinate the grass carp by oral route, the silkworm pupae expressing the rVP35-VP4 proteins were converted into a powder after freeze-drying, added to artificial feed at 5% and fed to grass carp (18 ± 1.5 g) for six weeks, and the immune response and protective efficacy in grass carp after oral vaccination trial was thoroughly investigated. This included blood cell counting and classification, serum antibody titer detection, immune-related gene expression and the relative percent survival rate in immunized grass carp. The results of blood cell counts show that the number of white blood cells in the peripheral blood of immunized grass carp increased significantly from 14 to 28 days post-immunization (dpi). The differential leukocyte count of neutrophils and monocytes were significantly higher than those in the control group at 14 dpi. Additionally, the number of lymphocytes increased significantly and reached a peak at 28 dpi. The serum antibody levels were significantly increased at Day 14 and continued until 42 days post-vaccination. The mRNA expression levels of immune-related genes (IFN-1, TLR22, IL-1β, MHC I, Mx and IgM) were significantly upregulated in liver, spleen, kidney and hindgut after immunization. Four weeks post-immunization, fish were challenged with virulent GCRV by intraperitoneal injection. The results of this challenge study show that orally immunized group exhibited a survival rate of 60% and relative percent survival (RPS) of 56%, whereas the control group had a survival rate of 13% and RPS of 4%. Taken together, our results demonstrate that the silkworm pupae powder containing baculovirus-expressed VP35-VP4 proteins could induce both non-specific and specific immune responses and protect grass carp against GCRV infection, suggesting it could be used as an oral vaccine.
Changyong Mu; Qiwang Zhong; Yan Meng; Yong Zhou; Nan Jiang; Wenzhi Liu; Yiqun Li; Mingyang Xue; Lingbing Zeng; Vikram N. Vakharia; Yuding Fan. Oral Vaccination of Grass Carp (Ctenopharyngodon idella) with Baculovirus-Expressed Grass Carp Reovirus (GCRV) Proteins Induces Protective Immunity against GCRV Infection. Vaccines 2021, 9, 41 .
AMA StyleChangyong Mu, Qiwang Zhong, Yan Meng, Yong Zhou, Nan Jiang, Wenzhi Liu, Yiqun Li, Mingyang Xue, Lingbing Zeng, Vikram N. Vakharia, Yuding Fan. Oral Vaccination of Grass Carp (Ctenopharyngodon idella) with Baculovirus-Expressed Grass Carp Reovirus (GCRV) Proteins Induces Protective Immunity against GCRV Infection. Vaccines. 2021; 9 (1):41.
Chicago/Turabian StyleChangyong Mu; Qiwang Zhong; Yan Meng; Yong Zhou; Nan Jiang; Wenzhi Liu; Yiqun Li; Mingyang Xue; Lingbing Zeng; Vikram N. Vakharia; Yuding Fan. 2021. "Oral Vaccination of Grass Carp (Ctenopharyngodon idella) with Baculovirus-Expressed Grass Carp Reovirus (GCRV) Proteins Induces Protective Immunity against GCRV Infection." Vaccines 9, no. 1: 41.
The grass carp hemorrhagic disease, caused by the grass carp reovirus (GCRV), has resulted in severe economic losses in the aquaculture industry in China. VP4 and VP35 are outer capsid proteins of GCRV and can induce an immune response in the host. Here, three recombinant baculoviruses, AcMNPV-VP35, AcMNPV-VP4, and AcMNPV-VP35-VP4, were generated to express recombinant VP4 and VP35 proteins from GCRV type II in insect cells by using the Bac-to-Bac baculovirus expression system to create a novel subunit vaccine. The expression of recombinant VP35, VP4, and VP35-VP4 proteins in Sf-9 cells were confirmed by Western blotting and immunofluorescence. Recombinant VP35, VP4, and VP35-VP4 were purified from baculovirus-infected cell lysates and injected intraperitoneally (3 μg/fish) into the model rare minnow, Gobiocypris rarus. After 21 days, the immunized fish were challenged with virulent GCRV. Liver, spleen, and kidney samples were collected at different time intervals to evaluate the protective efficacy of the subunit vaccines. The mRNA expression levels of some immune-related genes detected by using quantitative real-time PCR (qRT-PCR) were significantly upregulated in the liver, spleen, and kidney, with higher expression levels in the VP35-VP4 group. The nonvaccinated fish group showed 100% mortality, whereas the VP35-VP4, VP4, and VP35 groups exhibited 67%, 60%, and 33% survival, respectively. In conclusion, our results revealed that recombinant VP35 and VP4 can induce immunity and protect against GCRV infection, with their combined use providing the best effect. Therefore, VP35 and VP4 proteins can be used as a novel subunit vaccine against GCRV infection.
Changyong Mu; Vikram N. Vakharia; Yong Zhou; Nan Jiang; Wenzhi Liu; Yan Meng; Yiqun Li; Mingyang Xue; Jieming Zhang; Lingbing Zeng; Qiwang Zhong; Yuding Fan. A Novel Subunit Vaccine Based on Outer Capsid Proteins of Grass Carp Reovirus (GCRV) Provides Protective Immunity against GCRV Infection in Rare Minnow (Gobiocypris rarus). Pathogens 2020, 9, 945 .
AMA StyleChangyong Mu, Vikram N. Vakharia, Yong Zhou, Nan Jiang, Wenzhi Liu, Yan Meng, Yiqun Li, Mingyang Xue, Jieming Zhang, Lingbing Zeng, Qiwang Zhong, Yuding Fan. A Novel Subunit Vaccine Based on Outer Capsid Proteins of Grass Carp Reovirus (GCRV) Provides Protective Immunity against GCRV Infection in Rare Minnow (Gobiocypris rarus). Pathogens. 2020; 9 (11):945.
Chicago/Turabian StyleChangyong Mu; Vikram N. Vakharia; Yong Zhou; Nan Jiang; Wenzhi Liu; Yan Meng; Yiqun Li; Mingyang Xue; Jieming Zhang; Lingbing Zeng; Qiwang Zhong; Yuding Fan. 2020. "A Novel Subunit Vaccine Based on Outer Capsid Proteins of Grass Carp Reovirus (GCRV) Provides Protective Immunity against GCRV Infection in Rare Minnow (Gobiocypris rarus)." Pathogens 9, no. 11: 945.
Viral hemorrhagic septicemia virus (VHSV) is one of the most deadly infectious fish pathogens, posing a serious threat to the aquaculture industry and freshwater ecosystems worldwide. Previous work showed that VHSV sub-genotype IVb suppresses host innate immune responses, but the exact mechanism by which VHSV IVb inhibits antiviral response remains incompletely characterized. As with other novirhabdoviruses, VHSV IVb contains a unique and highly variable nonvirion (NV) gene, which is implicated in viral replication, virus-induced apoptosis and regulating interferon (IFN) production. However, the molecular mechanisms underlying the role of IVb NV gene in regulating viral or cellular processes is poorly understood. Compared to the wild-type recombinant (rWT) VHSV, mutant VHSV lacking a functional IVb NV reduced IFN expression and compromised innate immune response of the host cells by inhibiting translation. VHSV IVb infection increased phosphorylated eukaryotic initiation factor 2α (p-eIF2α), resulting in host translation shutoff. However, VHSV IVb protein synthesis proceeds despite increasing phosphorylation of eIF2α. During VHSV IVb infection, eIF2α phosphorylation was mediated via PKR-like endoplasmic reticulum kinase (PERK) and was required for efficient viral protein synthesis, but shutoff of host translation and IFN signaling was independent of p-eIF2α. Similarly, IVb NV null VHSV infection induced less p-eIF2α, but exhibited decreased viral protein synthesis despite increased levels of viral mRNA. These findings show a role for IVb NV in VHSV pathogenesis by utilizing the PERK-eIF2α pathway for viral-mediated host shutoff and interferon signaling to regulate host cell response.
Shelby Powell Kesterson; Jeffery Ringiesn; Vikram N. Vakharia; Brian S. Shepherd; Douglas W. Leaman; Krishnamurthy Malathi. Effect of the Viral Hemorrhagic Septicemia Virus Nonvirion Protein on Translation via PERK-eIF2α Pathway. Viruses 2020, 12, 499 .
AMA StyleShelby Powell Kesterson, Jeffery Ringiesn, Vikram N. Vakharia, Brian S. Shepherd, Douglas W. Leaman, Krishnamurthy Malathi. Effect of the Viral Hemorrhagic Septicemia Virus Nonvirion Protein on Translation via PERK-eIF2α Pathway. Viruses. 2020; 12 (5):499.
Chicago/Turabian StyleShelby Powell Kesterson; Jeffery Ringiesn; Vikram N. Vakharia; Brian S. Shepherd; Douglas W. Leaman; Krishnamurthy Malathi. 2020. "Effect of the Viral Hemorrhagic Septicemia Virus Nonvirion Protein on Translation via PERK-eIF2α Pathway." Viruses 12, no. 5: 499.
Rainbow trout farming is a major food source industry worldwide that has suffered great economic losses due to host jumps of fish rhabdovirus pathogens, followed by evolution of dramatic increases in trout-specific virulence. However, the genetic determinants of host jumps and increased virulence in rainbow trout are unknown for any fish rhabdovirus. Previous attempts to identify the viral genes containing trout virulence determinants of viral hemorrhagic septicemia virus (VHSV) have not been successful. We show here that, somewhat surprisingly, the viral nucleocapsid (N) and phosphoprotein (P) genes together contain the determinants responsible for trout virulence in VHSV. This suggests a novel host-specific virulence mechanism involving the viral polymerase and a host component. This differs from the known virulence mechanisms of mammalian rhabdoviruses based on the viral P or M (matrix) protein.
Vikram N. Vakharia; Jie Li; Douglas G. McKenney; Gael Kurath. The Nucleoprotein and Phosphoprotein Are Major Determinants of the Virulence of Viral Hemorrhagic Septicemia Virus in Rainbow Trout. Journal of Virology 2019, 93, 1 .
AMA StyleVikram N. Vakharia, Jie Li, Douglas G. McKenney, Gael Kurath. The Nucleoprotein and Phosphoprotein Are Major Determinants of the Virulence of Viral Hemorrhagic Septicemia Virus in Rainbow Trout. Journal of Virology. 2019; 93 (18):1.
Chicago/Turabian StyleVikram N. Vakharia; Jie Li; Douglas G. McKenney; Gael Kurath. 2019. "The Nucleoprotein and Phosphoprotein Are Major Determinants of the Virulence of Viral Hemorrhagic Septicemia Virus in Rainbow Trout." Journal of Virology 93, no. 18: 1.
Newcastle disease (ND) and avian reovirus (ARV) infections are a serious threat to the poultry industry, which causes heavy economic losses. The mesogenic NDV strain R2B is commonly used as a booster vaccine in many Asian countries to control the disease. In this seminal work, a recombinant NDV strain R2B expressing the sigma C (σC) gene of ARV (rNDV-R2B-σC) was generated by reverse genetics, characterized in vitro and tested as a bivalent vaccine candidate in chickens. The recombinant rNDV-R2B-σC virus was attenuated as compared to the parent rNDV-R2B virus as revealed by standard pathogenicity assays. The generated vaccine candidate, rNDV-R2B-σC, could induce both humoral and cell mediated immune responses in birds and gave complete protection against virulent NDV and ARV challenges. Post-challenge virus shedding analysis revealed a drastic reduction in NDV shed, as compared to unvaccinated birds.
Deep Prakash Saikia; Kalpana Yadav; Dinesh C. Pathak; Narayan Ramamurthy; Ajai Lawrence D’Silva; Asok Kumar Marriappan; Saravanan Ramakrishnan; Vikram N. Vakharia; Madhan Mohan Chellappa; Sohini Dey. Recombinant Newcastle Disease Virus (NDV) Expressing Sigma C Protein of Avian Reovirus (ARV) Protects against Both ARV and NDV in Chickens. Pathogens 2019, 8, 145 .
AMA StyleDeep Prakash Saikia, Kalpana Yadav, Dinesh C. Pathak, Narayan Ramamurthy, Ajai Lawrence D’Silva, Asok Kumar Marriappan, Saravanan Ramakrishnan, Vikram N. Vakharia, Madhan Mohan Chellappa, Sohini Dey. Recombinant Newcastle Disease Virus (NDV) Expressing Sigma C Protein of Avian Reovirus (ARV) Protects against Both ARV and NDV in Chickens. Pathogens. 2019; 8 (3):145.
Chicago/Turabian StyleDeep Prakash Saikia; Kalpana Yadav; Dinesh C. Pathak; Narayan Ramamurthy; Ajai Lawrence D’Silva; Asok Kumar Marriappan; Saravanan Ramakrishnan; Vikram N. Vakharia; Madhan Mohan Chellappa; Sohini Dey. 2019. "Recombinant Newcastle Disease Virus (NDV) Expressing Sigma C Protein of Avian Reovirus (ARV) Protects against Both ARV and NDV in Chickens." Pathogens 8, no. 3: 145.
Viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus belonging to the Novirhabdovirus genus, causes severe disease and mortality in many marine and freshwater fish species worldwide. VHSV isolates are classified into four genotypes and each group is endemic to specific geographic regions in the north Atlantic and Pacific Oceans. Most viruses in the European VHSV genotype Ia are highly virulent for rainbow trout (Oncorhynchus mykiss), whereas, VHSV genotype IVb viruses from the Great Lakes region in the United States, which caused high mortality in wild freshwater fish species, are avirulent for trout. This study describes molecular characterization and construction of an infectious clone of the virulent VHSV-Ia strain DK-3592B from Denmark, and application of the clone in reverse genetics to investigate the role of selected VHSV protein(s) in host-specific virulence in rainbow trout (referred to as trout-virulence). Overlapping cDNA fragments of the DK-3592B genome were cloned after RT-PCR amplification, and their DNA sequenced by the di-deoxy chain termination method. A full-length cDNA copy (pVHSVdk) of the DK-3592B strain genome was constructed by assembling six overlapping cDNA fragments by using natural or artificially created unique restriction sites in the overlapping regions of the clones. Using an existing clone of the trout-avirulent VHSV-IVb strain MI03 (pVHSVmi), eight chimeric VHSV clones were constructed in which the coding region(s) of the glycoprotein (G), non-virion protein (NV), G and NV, or G, NV and L (polymerase) genes together, were exchanged between the two clones. Ten recombinant VHSVs (rVHSVs) were generated, including two parental rVHSVs, by transfecting fish cells with ten individual full-length plasmid constructs along with supporting plasmids using the established protocol. Recovered rVHSVs were characterized for viability and growth in vitro and used to challenge groups of juvenile rainbow trout by intraperitoneal injection. Complete sequence of the VHSV DK-3592B genome was determined from the cloned cDNA and deposited in GenBank under the accession no. KC778774. The trout-virulent DK-3592B genome (genotype Ia) is 11,159 nt in length and differs from the trout-avirulent MI03 genome (pVHSVmi) by 13% at the nucleotide level. When the rVHSVs were assessed for the trout-virulence phenotype in vivo, the parental rVHSVdk and rVHSVmi were virulent and avirulent, respectively, as expected. Four chimeric rVHSVdk viruses with the substitutions of the G, NV, G and NV, or G, NV and L genes from the avirulent pVHSVmi constructs were still highly virulent (100% mortality), while the reciprocal four chimeric rVHSVmi viruses with genes from pVHSVdk remained avirulent (0–10% mortality). When chimeric rVHSVs, containing all the G, NV, and L gene substitutions, were tested in vivo, they did not exhibit any change in trout-virulence relative to the background clones. These results demonstrate that the G, NV and L genes of VHSV are not, by themselves or in combination, major determinants of host-specific virulence in trout.
Shamila Yusuff; Gael Kurath; Min Sun Kim; Tarin M. Tesfaye; Jie Li; Douglas G. McKenney; Vikram N. Vakharia. The glycoprotein, non-virion protein, and polymerase of viral hemorrhagic septicemia virus are not determinants of host-specific virulence in rainbow trout. Virology Journal 2019, 16, 1 -16.
AMA StyleShamila Yusuff, Gael Kurath, Min Sun Kim, Tarin M. Tesfaye, Jie Li, Douglas G. McKenney, Vikram N. Vakharia. The glycoprotein, non-virion protein, and polymerase of viral hemorrhagic septicemia virus are not determinants of host-specific virulence in rainbow trout. Virology Journal. 2019; 16 (1):1-16.
Chicago/Turabian StyleShamila Yusuff; Gael Kurath; Min Sun Kim; Tarin M. Tesfaye; Jie Li; Douglas G. McKenney; Vikram N. Vakharia. 2019. "The glycoprotein, non-virion protein, and polymerase of viral hemorrhagic septicemia virus are not determinants of host-specific virulence in rainbow trout." Virology Journal 16, no. 1: 1-16.
White Tail Disease (WTD) is one of the important viral diseases of fresh water giant prawn Macrobrachium rosenbergii, which is caused by Macrobrachium rosenbergii nodavirus (MrNV). In the present study, the capsid protein gene of MrNV containing a His-tag was cloned into a baculovirus vector pVL1393 and expressed the recombinant MrNV protein in insect cells, using a baculovirus expression system. A band corresponding to the MrNV protein of 43 kDa was characterized after fractionating the proteins of baculovirus-infected cell lysates by SDS-polyacrylamide gel, and immunostaining with His-tag monoclonal antibody. Furthermore, purified MrNV capsid protein assembled into virus-like particles (VLPs) of ∼30 nm in diameter, when examined by transmission electron microscopy (TEM). To vaccinate the larvae by oral route, the recombinant MrNV (r-MrNV) protein was coated with artificial prawn feed and fed to M. rosenbergii larvae (90 ± 10 mg) for 60 days. After 30 and 60 days of vaccine treatment, group of prawns were challenged with virulent MrNV orally. Samples were collected at different time intervals to evaluate the survival of larvae and to analyze the presence of MrNV by double-step PCR and expression of immune/ toll-like receptor (TLR) genes. Non-vaccinated group of M. rosenbergii larvae succumbed to death and had 90% mortality, whereas the r-MrNV protein treated groups exhibited 65 and 80% survival (P ≤ 0.001) for 30 and 60 days post-vaccination (dpv), respectively. Double-step PCR diagnosis revealed that there was 100% positive signals observed in non-vaccinated prawn group, whereas the infection was reduced significantly (P < 0.001) to 32 and 17% respectively in 30 and 60 dpv. Among the four different immune/ TLR genes such as antimicrobial peptide (Mramp), lysozyme (MrLY), proPhenol Oxidase (MrPPO) and Toll-Like Receptor (MrToll) expression screening, Mramp was successfully expressed in the MrNV subunit protein vaccinated prawns, whereas the non-vaccinated prawn had no immune/TLR gene expression. Taken together, our results demonstrate that oral vaccination of M. rosenbergii larvae with baculovirus-expressed MrNV capsid protein confer up to 78% protection against MrNV infection.
Thavasimuthu Citarasu; Chinnadurai Lelin; Mariavincent Michael Babu; Setty Balakrishnan Anand; Abel Arul Nathan; Vikram N. Vakharia. Oral vaccination of Macrobrachium rosenbergii with baculovirus-expressed M. rosenbergii nodavirus (MrNV) capsid protein induces protective immunity against MrNV challenge. Fish & Shellfish Immunology 2018, 86, 1123 -1129.
AMA StyleThavasimuthu Citarasu, Chinnadurai Lelin, Mariavincent Michael Babu, Setty Balakrishnan Anand, Abel Arul Nathan, Vikram N. Vakharia. Oral vaccination of Macrobrachium rosenbergii with baculovirus-expressed M. rosenbergii nodavirus (MrNV) capsid protein induces protective immunity against MrNV challenge. Fish & Shellfish Immunology. 2018; 86 ():1123-1129.
Chicago/Turabian StyleThavasimuthu Citarasu; Chinnadurai Lelin; Mariavincent Michael Babu; Setty Balakrishnan Anand; Abel Arul Nathan; Vikram N. Vakharia. 2018. "Oral vaccination of Macrobrachium rosenbergii with baculovirus-expressed M. rosenbergii nodavirus (MrNV) capsid protein induces protective immunity against MrNV challenge." Fish & Shellfish Immunology 86, no. : 1123-1129.
Although typical Newcastle disease virus (NDV) vaccines can prevent mortality, they are not effective preventing viral shedding. To overcome this, genotype-matched vaccines have been proposed. To date, this approach has never been tested against genotype XII strains. In this study, we generated and assessed the protection against genotype XII challenge of two chimeric NDV vaccine strains (rLS1-XII-1 and rLS1-XII-2). The rLS1-XII-1 virus has the complete fusion protein (F) and the hemmaglutinin-neuraminidase (HN) open reading frames replaced with those from genotype XII strain NDV/peacock/Peru/2011 (PP2011) in a recombinant LaSota (rLS1) backbone. For rLS1-XII-2 cytoplasmic tails of F and HN proteins were restored to those of rLS1. In vitro studies showed that rLS1-XII-2 and the parental rLS1 strains replicate at higher efficiencies than rLS1-XII-1. In the first vaccine/challenge experiment, SPF chickens vaccinated with rLS1-XII-1 virus showed only 71.3% protection, whereas, rLS1 and rLS1-XII-2 vaccinated chickens were fully protected. In a second experiment, both rLS1-XII-2 and the commercial vaccine strain LaSota induced 100% protection. However, rLS1-XII-2 virus significantly reduced viral shedding, both in the number of shedding birds and in quantity of shed virus. In conclusion, we have developed a vaccine candidate capable of fully protecting chickens against genotype XII challenges. Furthermore, we have shown the importance of cytoplasmic tails in virus replication and vaccine competence.
Ray Izquierdo-Lara; Ana Chumbe; Katherine Calderón; Manolo Fernández-Díaz; Vikram N Vakharia. Genotype-matched Newcastle disease virus vaccine confers improved protection against genotype XII challenge: The importance of cytoplasmic tails in viral replication and vaccine design. 2018, 492421 .
AMA StyleRay Izquierdo-Lara, Ana Chumbe, Katherine Calderón, Manolo Fernández-Díaz, Vikram N Vakharia. Genotype-matched Newcastle disease virus vaccine confers improved protection against genotype XII challenge: The importance of cytoplasmic tails in viral replication and vaccine design. . 2018; ():492421.
Chicago/Turabian StyleRay Izquierdo-Lara; Ana Chumbe; Katherine Calderón; Manolo Fernández-Díaz; Vikram N Vakharia. 2018. "Genotype-matched Newcastle disease virus vaccine confers improved protection against genotype XII challenge: The importance of cytoplasmic tails in viral replication and vaccine design." , no. : 492421.
Newcastle disease virus (NDV) strain F is a lentogenic vaccine strain used for primary vaccination in day-old chickens against Newcastle disease (ND) in India and Southeast Asian countries. Recombinant NDV-F virus and another recombinant NDV harboring the major capsid protein VP2 gene of a very virulent infectious bursal disease virus (IBDV); namely rNDV-F and rNDV-F/VP2, respectively, were generated using the NDV F strain. The rNDV-F/VP2 virus was slightly attenuated, as compared to the rNDV-F virus, as evidenced from the mean death time and intracerebral pathogenicity index analysis. This result indicates that rNDV-F/VP2 behaves as a lentogenic virus and it is stable even after 10 serial passages in embryonated chicken eggs. When chickens were vaccinated with the rNDV F/VP2, it induced both humoral and cell mediated immunity, and was able to confer complete protection against very virulent IBDV challenge and 80% protection against virulent NDV challenge. These results suggest that rNDV-F could be an effective and inherently safe vaccine vector. Here, we demonstrate that a bivalent NDV-IBDV vaccine candidate generated by reverse genetics method is safe, efficacious and cost-effective, which will greatly aid the poultry industry in developing countries.
Sohini Dey; Madhan Mohan Chellappa; Dinesh C. Pathak; Satish Gaikwad; Kalpana Yadav; Saravanan Ramakrishnan; Vikram N. Vakharia. Newcastle Disease Virus Vectored Bivalent Vaccine against Virulent Infectious Bursal Disease and Newcastle Disease of Chickens. Vaccines 2017, 5, 31 .
AMA StyleSohini Dey, Madhan Mohan Chellappa, Dinesh C. Pathak, Satish Gaikwad, Kalpana Yadav, Saravanan Ramakrishnan, Vikram N. Vakharia. Newcastle Disease Virus Vectored Bivalent Vaccine against Virulent Infectious Bursal Disease and Newcastle Disease of Chickens. Vaccines. 2017; 5 (4):31.
Chicago/Turabian StyleSohini Dey; Madhan Mohan Chellappa; Dinesh C. Pathak; Satish Gaikwad; Kalpana Yadav; Saravanan Ramakrishnan; Vikram N. Vakharia. 2017. "Newcastle Disease Virus Vectored Bivalent Vaccine against Virulent Infectious Bursal Disease and Newcastle Disease of Chickens." Vaccines 5, no. 4: 31.
MicroRNAs (miRNAs) play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV) has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1) with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi), respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b) on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future.
Xiaodan Liu; Jiagang Tu; Junfa Yuan; Xueqin Liu; Lijuan Zhao; Farman Ullah Dawar; Muhammad Nasir Khan Khattak; Abeer M. Hegazy; Nan Chen; Vikram N. Vakharia; Li Lin. Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection. International Journal of Molecular Sciences 2016, 17, 154 .
AMA StyleXiaodan Liu, Jiagang Tu, Junfa Yuan, Xueqin Liu, Lijuan Zhao, Farman Ullah Dawar, Muhammad Nasir Khan Khattak, Abeer M. Hegazy, Nan Chen, Vikram N. Vakharia, Li Lin. Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection. International Journal of Molecular Sciences. 2016; 17 (2):154.
Chicago/Turabian StyleXiaodan Liu; Jiagang Tu; Junfa Yuan; Xueqin Liu; Lijuan Zhao; Farman Ullah Dawar; Muhammad Nasir Khan Khattak; Abeer M. Hegazy; Nan Chen; Vikram N. Vakharia; Li Lin. 2016. "Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection." International Journal of Molecular Sciences 17, no. 2: 154.
Infectious bursal disease (IBD) is an acute, infectious, immunosuppressive disease affecting young chicken worldwide. The etiological agent IBD virus (IBDV) is a double stranded RNA virus with outer capsid protein VP2 of IBDV is the major antigenic determinant capable of inducing neutralizing antibody. DNA vaccines encoding VP2 has been extensively studied achieving only partial protection. However, the efficacy of DNA vaccines against IBDV can be augmented by choosing a potential molecular adjuvant. The goal of the present study is to evaluate the immune response and protective efficacy of a DNA vaccine encoding the C-terminal domain of the heat shock protein 70 (cHSP70) of Mycobacterium tuberculosis gene genetically fused with the full length VP2 gene of IBDV (pCIVP2-cHSP70) in comparison to a 'DNA prime-protein boost' approach and a DNA vaccine encoding the VP2 gene (pCIVP2) alone. The results indicate that both pCIVP2-cHSP70 and 'DNA prime-protein boost' elicited humoral as well as cellular immune responses. Chickens in the pCIVP2-cHSP70 and 'DNA prime-protein boost' groups developed significantly higher levels of ELISA titer to IBDV antigen compared to the group immunized with pCIVP2 alone (p<0.01). However, significantly higher levels of lymphocyte proliferative response, IL-12 and IFN-γ production were found in the pCIVP2-cHSP70 group compared to 'DNA prime-protein boost' group. Additionally, chickens immunized with pCIVP2-cHSP70 and 'DNA prime-protein boost' vaccines were completely protected against the vvIBDV whereas pCIVP2 DNA vaccine alone was able to protect only 70%. These findings suggest that the truncated C-terminal HSP70 mediated DNA vaccine genetically fused with the VP2 gene construct stimulated both humoral and cell mediated immune responses and conferred complete protection against IBDV. This novel strategy is perhaps a seminal concept in utilizing HSP70 as an adjuvant molecule to elicit an immune response against IBD affecting chickens.
Hemanta Maity; Sohini Dey; C. Madhan Mohan; Sagar A. Khulape; Dinesh C. Pathak; Vikram N. Vakharia. Protective efficacy of a DNA vaccine construct encoding the VP2 gene of infectious bursal disease and a truncated HSP70 of Mycobacterium tuberculosis in chickens. Vaccine 2015, 33, 1033 -1039.
AMA StyleHemanta Maity, Sohini Dey, C. Madhan Mohan, Sagar A. Khulape, Dinesh C. Pathak, Vikram N. Vakharia. Protective efficacy of a DNA vaccine construct encoding the VP2 gene of infectious bursal disease and a truncated HSP70 of Mycobacterium tuberculosis in chickens. Vaccine. 2015; 33 (8):1033-1039.
Chicago/Turabian StyleHemanta Maity; Sohini Dey; C. Madhan Mohan; Sagar A. Khulape; Dinesh C. Pathak; Vikram N. Vakharia. 2015. "Protective efficacy of a DNA vaccine construct encoding the VP2 gene of infectious bursal disease and a truncated HSP70 of Mycobacterium tuberculosis in chickens." Vaccine 33, no. 8: 1033-1039.
Viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) are members of the genus Novirhabdovirus within the Rhabdoviridae family, which can cause severe hemorrhagic disease in fresh- and saltwater fish worldwide. These viruses carry an additional nonvirion (NV) gene, which codes for the nonstructural NV protein that has been implicated to play a role in viral pathogenesis. To determine the precise biological function of this NV gene and its gene product, we generated NV-deficient and NV knockout recombinant VHSVs, using reverse genetics. Comparisons of the replication kinetics and markers for virus-induced apoptosis indicated that the NV-deficient and NV knockout mutant viruses induce apoptosis earlier in cell culture than the wild-type recombinant VHSV. These results suggest that the NV protein has an antiapoptotic function at the early stage of virus infection. Furthermore, we created a chimeric VHSV, in which the NV gene of VHSV was replaced by the IHNV NV gene, which was capable of suppressing apoptosis in cell culture. These results show that the NV protein of other members of Novirhabdovirus can restore the NV protein function. In this study, we also investigated the kinetics of VHSV replication during a single round of viral replication and examined the mechanism of VHSV-induced apoptosis. Our results show that VHSV infection induced caspases 3, 8 and 9 in cell culture.
Arun Ammayappan; Vikram N. Vakharia. Nonvirion Protein of Novirhabdovirus Suppresses Apoptosis at the Early Stage of Virus Infection. Journal of Virology 2011, 85, 8393 -8402.
AMA StyleArun Ammayappan, Vikram N. Vakharia. Nonvirion Protein of Novirhabdovirus Suppresses Apoptosis at the Early Stage of Virus Infection. Journal of Virology. 2011; 85 (16):8393-8402.
Chicago/Turabian StyleArun Ammayappan; Vikram N. Vakharia. 2011. "Nonvirion Protein of Novirhabdovirus Suppresses Apoptosis at the Early Stage of Virus Infection." Journal of Virology 85, no. 16: 8393-8402.
The routine technique for detecting antibodies specific to infectious bursal disease virus is a serological evaluation by enzyme linked immunosorbent assay (ELISA) with preparations of whole virions as antigens. To avoid the use of complete virus in the standard technique, in-house VP2 and VP3 based ELISAs were developed. Accordingly, four types of indirect ELISAs viz ., a commercial IDEXX-ELISA kit, VP2 and or VP3 antigen based ELISAs and a whole virus ELISA were compared with the virus neutralization test. It was concluded that the sensitivity and specificity at receiver-operating characteristics (ROC) optimized cut-off of four ELISAs viz ., IDEXX-ELISA, VP2-ELISA and VP3-ELISA indicated similar performance whereas whole virus antigen based ELISA showed poor performance in comparison to other ELISAs. Similarly the positive and negative likelihood ratio of four ELISAs at an optimized cut-off indicated IDEXX-ELISA to be the best among all the four ELISAs while the performance of rVP3-ELISA and rVP2-ELISA is good as compared to the whole virus ELISA. Finally, the area under the ROC curve (AUC) of four ELISAs which represented a summary statistics of the overall diagnostic performance of the test also indicated that the IDEXX-ELISA, VP3-ELISA and VP2-ELISA had similar and relatively better performance when compared to whole virus antigen-ELISA.
Niraj Kumar Singh; Sohini Dey; C. Madhan Mohan; Jag Mohan Kataria; Vikram N. Vakharia. Evaluation of four enzyme linked immunosorbent assays for the detection of antibodies to infectious bursal disease in chickens. Journal of Virological Methods 2010, 165, 277 -282.
AMA StyleNiraj Kumar Singh, Sohini Dey, C. Madhan Mohan, Jag Mohan Kataria, Vikram N. Vakharia. Evaluation of four enzyme linked immunosorbent assays for the detection of antibodies to infectious bursal disease in chickens. Journal of Virological Methods. 2010; 165 (2):277-282.
Chicago/Turabian StyleNiraj Kumar Singh; Sohini Dey; C. Madhan Mohan; Jag Mohan Kataria; Vikram N. Vakharia. 2010. "Evaluation of four enzyme linked immunosorbent assays for the detection of antibodies to infectious bursal disease in chickens." Journal of Virological Methods 165, no. 2: 277-282.