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Eduardo Furtado Flores - One of the best experts on this subject based on the ideXlab platform.

  • Retraction Note to: Comparative analysis of the replication of Bovine Herpesvirus 1(BHV1) and BHV5 in Bovine-derived neuron-like cells
    Archives of virology, 2020
    Co-Authors: Tereza C. Cardoso, Helena Lage Ferreira, Roberto Gameiro, Lucas Hidenori Okamura, Bruna R. S. M. Oliveira, Ana Carolina G. Rosa, Eduardo Furtado Flores
    Abstract:

    Members of the subfamily Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as preferential sites for primary replication. However, Bovine Herpesvirus 5 (BoHV5) is neurotropic and neuroinvasive and responsible for meningoencephalitis in cattle and in animal models. A related virus, BoHV1 has also been occasionally implicated in natural cases of neurological infection and disease in cattle. The aim of the present study was to assess the in vitro effects of BoHV1 and BoHV5 replication in neuron-like cells. Overall, cytopathic effects, consisting of floating rounded cells, giant cells and monolayer lysis, induced by both viruses at 48 h postinfection (p.i.) resulted in a loss of cell viability and high virus titres (r = 0.978). The BoHV1 Cooper strain produced the lowest titres in neuron-like cells, although viral DNA was detected in infected cells during all experiments. Virus replication in infected cells was demonstrated by immunocytochemistry, flow cytometry and qPCR assays. BoHV antigens were better visualized at 48 h p.i. and flow cytometry analysis showed that SV56/90 and Los Angeles antigens were present at higher levels. In spite of the fact that BoHV titres dropped at 48 h p.i, viral DNA remained detectable until 120 h p.i. Sensitive TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and annexin V assays were used to identify apoptosis. BoHV5 induced death in approximately 50 % of cells within 24 h p.i., similar to what has been observed for BoHV1 Los Angeles. Infection with the BoHV1 Cooper strain resulted in 26.37 % of cells being in the early stages of apoptosis; 63.69 % of infected cells were considered viable. Modulation of mitochondrial function, as measured by mitochondrial membrane depolarization, was synchronous with the virus replication cycle, cell viability and virus titres at 48 h p.i. Our results indicate that apoptosis plays an important role in preventing neuronal death and provides a Bovine-derived in vitro system to study Herpesvirus-neuron interactions.

  • RETRACTED: Bovine Herpesvirus 5 promotes mitochondrial dysfunction in cultured Bovine monocyte-derived macrophages and not affect virus replication.
    Veterinary Microbiology, 2019
    Co-Authors: Tereza C. Cardoso, Lucas H. Okamura, Jamila C. Baptistella, Ana Carolina Borsanelli, Lillian Baptistiolli, Helena Lage Ferreira, Roberto Gameiro, Eduardo Furtado Flores
    Abstract:

    This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editors-in-Chief and Authors. Fig 1A is a duplicate of a figure that has already been published in da Silva SEL et al. Archives of Virology 2018;163:1043-1049; 10.1007/s00705-018-3704-2. These two papers report studies performed with cells from two different animal species (Bovine cells for the Veterinary Microbiology paper and chicken cells for the Archives of Virology paper). The reuse of the same figure in the Veterinary Microbiology paper to describe cells that were supposed to be from a different species is thus inappropriate and also puts into question the reliability of the other results presented in this paper. In addition, the Editors-in-Chief have remaining concerns about the strong similarities of other data presented in the two papers. Even if these concerns were addressed, the re-use of any data has to be clearly indicated and appropriately cited. As such this article represents a misuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

  • Cellular response markers and cytokine gene expression in the central nervous system of cattle naturally infected with Bovine Herpesvirus 5
    Veterinary journal (London England : 1997), 2016
    Co-Authors: Tereza C. Cardoso, Helena Lage Ferreira, Roberto Gameiro, Lucas Hidenori Okamura, T.p. Giroto, Bruna R. S. M. Oliveira, C.u.f. Fabri, Eduardo Furtado Flores
    Abstract:

    Abstract The present study reports an investigation on the phenotype of inflammatory and immune cells, cytokine and viral gene expression in the brains of cattle naturally infected with Bovine Herpesvirus 5 (BHV5). Brain sections of 38 affected animals were analysed for the nature and extent of perivascular cuffs in the Virchow–Robin space and parenchyma. Histopathological changes were severe in the olfactory bulbs (Obs), hippocampus, piriform, frontal, temporal and parietal cortices/lobes and were characterized by inflammatory infiltrates in Virchow–Robin spaces. The histopathological changes correlated positively with the distribution of BHV5 antigens ( r  = 0.947; P r  = 0.987; P INFG , IL2 , TNF and LTBR were expressed in the same brain areas ( P

  • Comparative analysis of the replication of Bovine Herpesvirus 1 (BHV1) and BHV5 in Bovine-derived neuron-like cells.
    Archives of virology, 2015
    Co-Authors: Tereza C. Cardoso, Helena Lage Ferreira, Roberto Gameiro, Lucas Hidenori Okamura, Bruna R. S. M. Oliveira, Ana Carolina G. Rosa, Eduardo Furtado Flores
    Abstract:

    Members of the subfamily Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as preferential sites for primary replication. However, Bovine Herpesvirus 5 (BoHV5) is neurotropic and neu- roinvasive and responsible for meningoencephalitis in cattle and in animal models. A related virus, BoHV1 has also been occasionally implicated in natural cases of neu- rological infection and disease in cattle. The aim of the present study was to assess the in vitro effects of BoHV1 and BoHV5 replication in neuron-like cells. Overall, cytopathic effects, consisting of floating rounded cells, giant cells and monolayer lysis, induced by both viruses at 48 h postinfection (p.i.) resulted in a loss of cell viability and high virus titres (r = 0.978). The BoHV1 Cooper strain produced the lowest titres in neuron-like cells, although viral DNA was detected in infected cells during all experiments. Virus replication in infected cells was demonstrated by immunocytochemistry, flow cytometry and qPCR assays. BoHV antigens were better visualized at 48 h p.i. and flow cytometry analysis showed that SV56/90 and Los Angeles antigens were present at higher levels. In spite of the fact that BoHV titres dropped at 48 h p.i, viral DNA remained detectable until 120 h p.i. Sensitive TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and annexin V assays were used to identify apoptosis. BoHV5 induced death in approximately 50 % of cells within 24 h p.i., similar to what has been observed for BoHV1 Los Angeles. Infection with the BoHV1 Cooper strain resulted in 26.37 % of cells being in the early stages of apoptosis; 63.69 % of infected cells were considered viable. Modulation of mitochondrial function, as measured by mitochondrial membrane depo- larization, was synchronous with the virus replication cycle, cell viability and virus titres at 48 h p.i. Our results indicate that apoptosis plays an important role in prevent- ing neuronal death and provides a Bovine-derived in vitro system to study Herpesvirus-neuron interactions.

  • A RETROSPECTIVE SEARCH FOR Bovine Herpesvirus 5 (BOHV-5) IN THE BRAIN OF CATTLE AFFECTED BY NEUROLOGICAL DISEASE IN RIO DE JANEIRO STATE
    VIRUS Reviews & Research, 2014
    Co-Authors: Ana Maria Viana Pinto, Eduardo Furtado Flores, R. Weiblen, José Paulo Gagliardi Leite, P. C. Romijn, R. C. F. Silva, Izabel Christina Nunes De Palmer Paixão
    Abstract:

    Bovine Herpesvirus 5 (BoHV-5) is an important agent of meningoencephalitis in cattle and has been identified in outbreaks of neurological disease in several Brazilian states. However, no report of BoHV-5-associated neurological disease in Rio Janeiro state (RJ) has been published to date. This article reports a retrospective investigation for BoHV-5 performed in 146 brain specimens of cattle affected by neurological disease, submitted to the virology laboratory of the Empresa de Pesquisa Agropecuaria do Estado do Rio de Janeiro (PESAGRO-RJ) between 1998 and 2001. Most specimens came from herds in the RJ state (130 or 89%) and a few from Minas Gerais counties (16 or 11%), close to the RJ border. All brain samples examined in this study were previously diagnosed negative for rabies virus. The specimens were submitted to PCR/nested-PCR for BoHV-5 sequences and virus isolation. From 146 analysed brains samples 48 (33%) were positive in at least one of the tests (PCR or virus isolation), 20 samples (14%) were positive in both assays, 19 samples (13%) were positive in Nested-PCR (n-PCR) and nine samples (6%) were characterized as BoHV-5 after virus isolation. The northeast region of the RJ contributed with most clinical specimens (92/146 or 63%) and presented the highest positivity (28/92 or 30%). These results demonstrated BoHV-5 association with neurological disease in Bovine herds of RJ and MG and should therefore be included in differential diagnosis of neurological disease in cattle. The limitations of each of the diagnostic assays used here in led us to recommend the concomitant use of PCR and virus isolation to improve the sensitivity of the diagnosis of BoHV-5 infection. DOI:  http://dx.doi.org/10.17525/vrrjournal.v19i2.140

Fábio Pereira Leivas Leite - One of the best experts on this subject based on the ideXlab platform.

  • Association of Bacillus toyonensis spores with alum improves Bovine Herpesvirus 5 subunit vaccine immune response in mice
    Vaccine, 2020
    Co-Authors: Lucas Reichert Maubrigades, Francisco Denis Souza Santos, Vitória Sequeira Gonçalves, Paulo Ricardo Centeno Rodrigues, Fábio Pereira Leivas Leite
    Abstract:

    Abstract Spores of the genus Bacillus are molecules capable of increasing the vaccine adjuvanticity. Bovine Herpesvirus type 5 (BoHV-5) is responsible for meningoencephalitis that causes important economic losses in cattle. BoHV-5 glycoprotein D (gD) is a target of vaccine antigen and plays an important role in host cell penetration. The present study aimed to evaluate the adjuvanticity of Bacillus toyonensis (B.t) spores, live and heat-killed, associated with a vaccine formulated with aluminum hydroxide (alum) and the recombinant BoHV-5 glycoprotein D (rgD) in an experimental murine model. Six experimental groups of mice were subcutaneously vaccinated on day 0 and received a booster on day 21 of the experiment, with the following vaccine formulations: rgD (40 µg) + live spores (2 × 109 CFU); rgD + killed spores; rgD + live spores + alum (2.0 mg); rgD + killed spores + alum; rgD + alum, and rgD + PBS. Mice from rgD + live spores group showed an increase in rgD IgG titers from the 21st day until the end of the experiment. The groups of live and killed spores, associated to alum, had similar levels of IgG titers with no significant difference between each other; however, by the 14th and 28th day until the end of the experiment, presented higher IgG titers in comparison to the rgD + alum group. Moreover, increased serum levels of IgG1, IgG2a, and IgG2b were detected in mice that received spores in the vaccine formulation. The spores associated with alum groups showed neutralizing BoHV-5 antibodies and high mRNA transcription of the cytokines IFN-γ (66-fold), IL-17 (14-fold), and IL-12 (2.8-fold). In conclusion, our data demonstrated that the B. toyonensis spores, live or killed, associated with alum increased the adjuvanticity for BoHV-5 rgD in mice, suggesting the use of B. toyonensis spores as a promising component for vaccine formulations.

  • immune responses in Bovines to recombinant glycoprotein d of Bovine Herpesvirus type 5 as vaccine antigen
    Vaccine, 2018
    Co-Authors: Itauá Leston Araujo, Luana Alves Dummer, Alceu Gonçalves Dos Santos, Geferson Fischer, Paulo Ricardo Centeno Rodrigues, Rodrigo Casquero Cunha, Fábio Pereira Leivas Leite
    Abstract:

    Bovine Herpesvirus 5 (BoHV-5) is responsible for outbreaks of meningoencephalitis that cause important economic losses in young cattle. BoHV-5 glycoprotein D (gD5) is essential for attachment and penetration into permissive cells and targeting of host immune systems, inducing strong humoral and cellular immune responses. The aim of this study was to evaluate the vaccinal immune response of vaccines formulated with the recombinant BoHV-5 gD (rgD5) in Bovines. For the experiment, 72 heifers were randomly allotted into 6 different groups with 12 animals each. Group 1: vaccine formulated using inactivated BoHV-5 (iBoHV-5) adjuvanted with ISA50V2; Group 2: iBoHV-5 associated with 100 µg of rgD5 adjuvanted with ISA50V2; Group 3: 100 µg of rgD5 adjuvanted with ISA50V2; Group 4: 100 µg of rgD5 adjuvanted with Al(OH)3; Group 5: commercial vaccine; and Group 6: control group. Two doses were administered in a 26-day interval and the third after 357 days from primo vaccination. Cattle vaccinated with the vaccines formulated with iBoHV-5 plus rgD5 showed a significant (p < 0.01) five-fold increase in total immunoglobulin G (IgG) for BoHV-5, BoHV-1, and rgD5 as compared with the commercial and control groups. Also, a significant (p < 0.05) increase in IgG1 and IgG2a levels was induced in serum for rgD5. In addition, these same vaccines showed significant (p < 0.01) four-fold higher titers of BoHV-1 and -5 neutralizing antibodies. The results demonstrated that the rgD5 conserved important epitopes that were able to stimulate Bovine humoral immunity response capable of viral neutralization of BoHV-1 and -5, suggesting it as a promising vaccine antigen to be used in vaccine for BoHV-1 and -5 endemic areas.

  • Immune responses in Bovines to recombinant glycoprotein D of Bovine Herpesvirus type 5 as vaccine antigen.
    Vaccine, 2018
    Co-Authors: Itauá Leston Araujo, Luana Alves Dummer, Alceu Gonçalves Dos Santos, Geferson Fischer, Paulo Ricardo Centeno Rodrigues, Rodrigo Casquero Cunha, Fábio Pereira Leivas Leite
    Abstract:

    Bovine Herpesvirus 5 (BoHV-5) is responsible for outbreaks of meningoencephalitis that cause important economic losses in young cattle. BoHV-5 glycoprotein D (gD5) is essential for attachment and penetration into permissive cells and targeting of host immune systems, inducing strong humoral and cellular immune responses. The aim of this study was to evaluate the vaccinal immune response of vaccines formulated with the recombinant BoHV-5 gD (rgD5) in Bovines. For the experiment, 72 heifers were randomly allotted into 6 different groups with 12 animals each. Group 1: vaccine formulated using inactivated BoHV-5 (iBoHV-5) adjuvanted with ISA50V2; Group 2: iBoHV-5 associated with 100 µg of rgD5 adjuvanted with ISA50V2; Group 3: 100 µg of rgD5 adjuvanted with ISA50V2; Group 4: 100 µg of rgD5 adjuvanted with Al(OH)3; Group 5: commercial vaccine; and Group 6: control group. Two doses were administered in a 26-day interval and the third after 357 days from primo vaccination. Cattle vaccinated with the vaccines formulated with iBoHV-5 plus rgD5 showed a significant (p 

  • Saccharomyces boulardii modulates and improves the immune response to Bovine Herpesvirus type 5 Vaccine
    Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 2018
    Co-Authors: Talita Bandeira Roos, Geferson Fischer, Luciana Farias Da Costa De Avila, R.t. Sturbelle, F.l.l. Leite, Fábio Pereira Leivas Leite
    Abstract:

    ABSTRACT There have been significant efforts towards the development of more efficient vaccines for animal health. A strategy that may be used to improve vaccine efficacy is the use of probiotics to enhance the immune response of the host, leading to increased immunogenicity of antigen preparations. Bovine Herpesvirus 5 (BoHV-5) is an example of an important animal pathogen for which vaccines have provided only limited protection. In this study, we examined the use of the probiotic Saccharomyces boulardii (Sb) as a potential adjuvant to improve vaccine efficiency. We found that the supplemented animals exhibited an enhanced systemic IgG antibody response toward a Th1 response in favor of IgG2a and increased mRNA expression levels of the cytokines IFN-y, IL-12, IL-17 and IL-10 in the spleen. These results suggest that Sb supplementation may provide a promising means for improving the efficiency of vaccines, particularly those that rely on a cell-mediated immune response.

  • Probiotics Bacillus toyonensis and Saccharomyces boulardii improve the vaccine immune response to Bovine Herpesvirus type 5 in sheep
    Research in veterinary science, 2017
    Co-Authors: Talita Bandeira Roos, Luana Alves Dummer, Geferson Fischer, R.t. Sturbelle, Carina Martins De Moraes, Fábio Pereira Leivas Leite
    Abstract:

    There have been significant efforts toward the development of more efficient vaccines for animal health. A strategy that may be used to improve vaccine efficacy is the use of probiotics. Bovine Herpesvirus 5 (BoHV-5) is an example of an important animal pathogen for which vaccines have provided only limited protection. In this study, we examined the use of the probiotics Bacillus toyonensis and Saccharomyces boulardii as a potential immune modulator to improve vaccine efficiency. Thirty, 5-month-old lambs were randomly grouped in three lots of 10 each and vaccinated at days 0, 21 and 42 of the experiment. They grazed on the same pasture and were fed ad libitum twice a day with commercial sheep feed supplemented with either B. toyonensis (1×106CFU/g of feed) or S. boulardii (1×107CFU/g of feed), or non-supplemented feed. The probiotic supplementation was suspended day 28; thereafter, the next 35days, they were fed with the same commercial feed as control group. Animals supplemented with probiotics showed a significant (p>0.001) increased seroconversions against BoHV-5, and higher neutralizing antibodies titres (p>0.05) to BoHV-5 than non-supplemented animals. At 63days of experiment, splenocytes from the supplemented sheep had higher mRNA transcription levels of cytokines IL-10 and IL-17A. These results suggest that these probiotics could provide a promising means of improving vaccine efficacy.

Tereza C. Cardoso - One of the best experts on this subject based on the ideXlab platform.

  • Retraction Note to: Comparative analysis of the replication of Bovine Herpesvirus 1(BHV1) and BHV5 in Bovine-derived neuron-like cells
    Archives of virology, 2020
    Co-Authors: Tereza C. Cardoso, Helena Lage Ferreira, Roberto Gameiro, Lucas Hidenori Okamura, Bruna R. S. M. Oliveira, Ana Carolina G. Rosa, Eduardo Furtado Flores
    Abstract:

    Members of the subfamily Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as preferential sites for primary replication. However, Bovine Herpesvirus 5 (BoHV5) is neurotropic and neuroinvasive and responsible for meningoencephalitis in cattle and in animal models. A related virus, BoHV1 has also been occasionally implicated in natural cases of neurological infection and disease in cattle. The aim of the present study was to assess the in vitro effects of BoHV1 and BoHV5 replication in neuron-like cells. Overall, cytopathic effects, consisting of floating rounded cells, giant cells and monolayer lysis, induced by both viruses at 48 h postinfection (p.i.) resulted in a loss of cell viability and high virus titres (r = 0.978). The BoHV1 Cooper strain produced the lowest titres in neuron-like cells, although viral DNA was detected in infected cells during all experiments. Virus replication in infected cells was demonstrated by immunocytochemistry, flow cytometry and qPCR assays. BoHV antigens were better visualized at 48 h p.i. and flow cytometry analysis showed that SV56/90 and Los Angeles antigens were present at higher levels. In spite of the fact that BoHV titres dropped at 48 h p.i, viral DNA remained detectable until 120 h p.i. Sensitive TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and annexin V assays were used to identify apoptosis. BoHV5 induced death in approximately 50 % of cells within 24 h p.i., similar to what has been observed for BoHV1 Los Angeles. Infection with the BoHV1 Cooper strain resulted in 26.37 % of cells being in the early stages of apoptosis; 63.69 % of infected cells were considered viable. Modulation of mitochondrial function, as measured by mitochondrial membrane depolarization, was synchronous with the virus replication cycle, cell viability and virus titres at 48 h p.i. Our results indicate that apoptosis plays an important role in preventing neuronal death and provides a Bovine-derived in vitro system to study Herpesvirus-neuron interactions.

  • RETRACTED: Bovine Herpesvirus 5 promotes mitochondrial dysfunction in cultured Bovine monocyte-derived macrophages and not affect virus replication.
    Veterinary Microbiology, 2019
    Co-Authors: Tereza C. Cardoso, Lucas H. Okamura, Jamila C. Baptistella, Ana Carolina Borsanelli, Lillian Baptistiolli, Helena Lage Ferreira, Roberto Gameiro, Eduardo Furtado Flores
    Abstract:

    This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editors-in-Chief and Authors. Fig 1A is a duplicate of a figure that has already been published in da Silva SEL et al. Archives of Virology 2018;163:1043-1049; 10.1007/s00705-018-3704-2. These two papers report studies performed with cells from two different animal species (Bovine cells for the Veterinary Microbiology paper and chicken cells for the Archives of Virology paper). The reuse of the same figure in the Veterinary Microbiology paper to describe cells that were supposed to be from a different species is thus inappropriate and also puts into question the reliability of the other results presented in this paper. In addition, the Editors-in-Chief have remaining concerns about the strong similarities of other data presented in the two papers. Even if these concerns were addressed, the re-use of any data has to be clearly indicated and appropriately cited. As such this article represents a misuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

  • Expression of miR-155 associated with Toll-like receptors 3, 7, and 9 transcription in the olfactory bulbs of cattle naturally infected with BHV5
    Journal of NeuroVirology, 2017
    Co-Authors: Bruna R. S. M. Oliveira, Roberto Gameiro, Flavia V. Vieira, Eduardo F. Flores, Dielson Vieira, Sergio E. L. Silva, Tereza C. Cardoso
    Abstract:

    Bovine Herpesvirus 5 (BHV5) infection of young cattle is frequently associated with fatal neurological disease and, as such, represents an attractive model for studying the pathogenesis of viral-induced meningoencephalitis. Following replication in the nasal mucosa, BHV5 invades the central nervous system (CNS) mainly through the olfactory pathway. The innate immune response triggered by the host face to virus replication through the olfactory route is poorly understood. Recently, an upregulation of conserved pathogen-associated molecular pattern, as Toll-like receptors (TLRs), has been demonstrated in the CNS of BHV5 experimentally infected cows. A new perspective to understand host-pathogen interactions has emerged elucidating microRNAs (miRNAs) network that interact with innate immune response during neurotropic viral infections. In this study, we demonstrated a link between the expression of TLRs 3, 7, and 9 and miR-155 transcription in the olfactory bulbs (OB) of 16 cows suffering from acute BHV5-induced neurological disease. The OBs were analyzed for viral antigens and genome, miR-155 and TLR 3, 7, and 9 expression considering three major regions: olfactory receptor neurons (ORNs), glomerular layer (GL), and mitral cell layer (ML). BHV5 antigens and viral genomes, corresponding to glycol-C gene, were detected in all OBs regions by fluorescent antibody assay (FA) and PCR, respectively. TLR 3, 7, and 9 transcripts were upregulated in ORNs and ML, yet only ORN layers revealed a positive correlation between TLR3 and miR-155 transcription. In ML, miR-155 correlated positively with all TLRs studied. Herein, our results evidence miR-155 transcription in BHV5 infected OB tissue associated to TLRs expression specifically ORNs which may be a new window for further studies.

  • Bovine Herpesviruses induce different cell death forms in neuronal and glial-derived tumor cell cultures
    Journal of NeuroVirology, 2016
    Co-Authors: Tereza C. Cardoso, Lucas H. Okamura, Roberto Gameiro, Bruna R. S. M. Oliveira, Ana Carolina G. Rosa, Helena L. Ferreira, Flavia V. Vieira, Camila Silva-frade, Eduardo F. Flores
    Abstract:

    Oncolytic viruses have the ability to infect tumor cells and leave healthy cells intact. In this study, Bovine Herpesvirus 1 (BHV1; Los Angeles, Cooper, and SV56/90 strains) and Bovine Herpesvirus 5 (BHV5; SV507/99 and GU9457818 strains) were used to infect two neuronal tumor cell lineages: neuro2a (mouse neuroblastoma cells) and C6 (rat glial cells). BHV1 and BHV5 strains infected both cell lines and positively correlated with viral antigen detection ( p  40 % in necroptosis phase when infected by BHV5 (GU9457818 strain). Blocking caspase activation did not interfere with cell death. However, when necroptosis was blocked, 60–80 % of both infected cells with either virus switched to early apoptosis pathway with no interference with virus replication. Moreover, reactive oxygen species production and mitochondrial membrane dysfunction were detected at high levels in both infected cell lines. In spite of apoptosis and necroptosis blockage, tumor necrosis factor alpha ( TNFA ) and virus transcription were positively correlated for all viral strains studied. Thus, these results contribute to the characterization of BHV1 and BHV5 as potential oncolytic viruses for non-human cells. Nonetheless, the mechanisms underlying their oncolytic activity in human cells are still to be determined.

  • Cellular response markers and cytokine gene expression in the central nervous system of cattle naturally infected with Bovine Herpesvirus 5
    Veterinary journal (London England : 1997), 2016
    Co-Authors: Tereza C. Cardoso, Helena Lage Ferreira, Roberto Gameiro, Lucas Hidenori Okamura, T.p. Giroto, Bruna R. S. M. Oliveira, C.u.f. Fabri, Eduardo Furtado Flores
    Abstract:

    Abstract The present study reports an investigation on the phenotype of inflammatory and immune cells, cytokine and viral gene expression in the brains of cattle naturally infected with Bovine Herpesvirus 5 (BHV5). Brain sections of 38 affected animals were analysed for the nature and extent of perivascular cuffs in the Virchow–Robin space and parenchyma. Histopathological changes were severe in the olfactory bulbs (Obs), hippocampus, piriform, frontal, temporal and parietal cortices/lobes and were characterized by inflammatory infiltrates in Virchow–Robin spaces. The histopathological changes correlated positively with the distribution of BHV5 antigens ( r  = 0.947; P r  = 0.987; P INFG , IL2 , TNF and LTBR were expressed in the same brain areas ( P

S I Chowdhury - One of the best experts on this subject based on the ideXlab platform.

  • A recombinant Bovine Herpesvirus 5 defective in thymidine kinase and glycoprotein E is immunogenic for calves and confers protection upon homologous challenge and BoHV-1 challenge.
    Veterinary microbiology, 2011
    Co-Authors: D Anziliero, R. Weiblen, C M B Santos, M C S Brum, S I Chowdhury, E. F. Flores
    Abstract:

    A recombinant Bovine Herpesvirus 5 lacking thymidine kinase and glycoprotein E genes (BoHV-5gEΔTKΔ) was evaluated as a live experimental vaccine. In a first experiment, ten-months-old calves were vaccinated intramuscularly (n=9) or remained as controls (n=8) and 42 days later were challenged with BoHV-5 or BoHV-1 intranasally. The four control calves challenged with BoHV-5 developed severe depression and neurological signs and were euthanized in extremis at days 13 and 14 pos-infection (pi); the five vaccinated animals challenged with BoHV-5 remained healthy. The titers of virus shedding were reduced (p

  • A recombinant Bovine Herpesvirus 5 defective in thymidine kinase and glycoprotein E is immunogenic for calves and confers protection upon homologous challenge and BoHV-1 challenge.
    Veterinary Microbiology, 2011
    Co-Authors: D Anziliero, Rudi Weiblen, C M B Santos, M C S Brum, S I Chowdhury, Eduardo Furtado Flores
    Abstract:

    Abstract A recombinant Bovine Herpesvirus 5 lacking thymidine kinase and glycoprotein E genes (BoHV-5gEΔTKΔ) was evaluated as a live experimental vaccine. In a first experiment, ten-months-old calves were vaccinated intramuscularly ( n  = 9) or remained as controls ( n  = 8) and 42 days later were challenged with BoHV-5 or BoHV-1 intranasally. The four control calves challenged with BoHV-5 developed severe depression and neurological signs and were euthanized in extremis at days 13 and 14 pos-infection (pi); the five vaccinated animals challenged with BoHV-5 remained healthy. The titers of virus shedding were reduced ( p p n  = 15) or kept as controls ( n  = 5) and subsequently challenged with a BoHV-1 isolate. Control calves developed moderate to severe rhinitis and respiratory distress; two were euthanized in extremis at days 5 and 9 pi, respectively. In contrast, vaccinated animals were protected from challenge and only a few developed mild and transient nasal signs. The duration and titers of virus shedding after challenge were reduced ( p

  • A Bovine Herpesvirus 5 recombinant defective in the thymidine kinase (TK) gene and a double mutant lacking TK and the glycoprotein E gene are fully attenuated for rabbits
    Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas, 2009
    Co-Authors: Sara C. Da Silva, Eduardo Furtado Flores, Rudi Weiblen, M C S Brum, S I Chowdhury
    Abstract:

    Bovine Herpesvirus 5 (BoHV-5), the agent of herpetic meningoencephalitis in cattle, is an important pathogen of cattle in South America and several efforts have been made to produce safer and more effective vaccines. In the present study, we investigated in rabbits the virulence of three recombinant viruses constructed from a neurovirulent Brazilian BoHV-5 strain (SV507/99). The recombinants are defective in glycoprotein E (BoHV-5gEΔ), thymidine kinase (BoHV-5TKΔ) and both proteins (BoHV-5gEΔTKΔ). Rabbits inoculated with the parental virus (N = 8) developed neurological disease and died or were euthanized in extremis between days 7 and 13 post-infection (pi). Infectivity was detected in several areas of their brains. Three of 8 rabbits inoculated with the recombinant BoHV-5gEΔ developed neurological signs between days 10 and 15 pi and were also euthanized. A more restricted virus distribution was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV-5TKΔ (N = 8) or BoHV-5gEΔTKΔ (N = 8) remained healthy throughout the experiment in spite of variable levels of virus replication in the nose. Dexamethasone (Dx) administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by absence of virus shedding and/or increase in virus neutralizing titers. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all animals at day 28 post-Dx, demonstrating they were latently infected. These results show that recombinants BoHV-5TKΔ and BoHV-5gEΔTKΔ are attenuated for rabbits and constitute potential vaccine candidates upon the confirmation of this phenotype in cattle.

  • Bovine Herpesvirus 5 (BHV-5) Us9 Is Essential for BHV-5 Neuropathogenesis
    Journal of virology, 2002
    Co-Authors: S I Chowdhury, M. Onderci, Partha S. Bhattacharjee, A. Al-mubarak, Mark L. Weiss, You Zhou
    Abstract:

    Bovine Herpesvirus 5 (BHV-5) is a neurovirulent alphaHerpesvirus that causes fatal encephalitis in calves. In a rabbit model, the virus invades the central nervous system (CNS) anterogradely from the olfactory mucosa following intranasal infection. In addition to glycoproteins E and I (gE and gI, respectively), Us9 and its homologue in alphaHerpesviruses are necessary for the viral anterograde spread from the presynaptic to postsynaptic neurons. The BHV-5 Us9 gene sequence was determined, and the predicted amino acid sequence of BHV-5 Us9 was compared with the corresponding Us9 sequences of BHV-1.1. Alignment results showed that they share 77% identity and 83% similarity. BHV-5 Us9 peptide-specific antibody recognized a doublet of 17- and 19-kDa protein bands in BHV-5-infected cell lysates and in purified virions. To determine the role of the BHV-5 Us9 gene in BHV-5 neuropathogenesis, a BHV-5 Us9 deletion recombinant was generated and its neurovirulence and neuroinvasive properties were compared with those of a Us9 rescue mutant of BHV-5 in a rabbit model. Following intranasal infection, the Us9 rescue mutant of BHV-5 displayed a wild-type level of neurovirulence and neural spread in the olfactory pathway, but the Us9 deletion mutant of BHV-5 was virtually avirulent and failed to invade the CNS. In the olfactory mucosa containing the olfactory receptor neurons, the Us9 deletion mutant virus replicated with an efficiency similar to that of the Us9 rescue mutant of BHV-5. However, the Us9 deletion mutant virus was not transported to the bulb. Confocal microscopy of the olfactory epithelium detected similar amounts of virus-specific antigens in the cell bodies of olfactory receptor neuron for both the viruses, but only the Us9 rescue mutant viral proteins were detected in the processes of the olfactory receptor neurons. When injected directly into the bulb, both viruses were equally neurovirulent, and they were transported retrogradely to areas connected to the bulb. Taken together, these results indicate that Us9 is essential for the anterograde spread of the virus from the olfactory mucosa to the bulb.

  • Bovine Herpesvirus 5 Glycoprotein E Is Important for Neuroinvasiveness and Neurovirulence in the Olfactory Pathway of the Rabbit
    Journal of virology, 2000
    Co-Authors: S I Chowdhury, Bong Joo Lee, Aykut Ozkul, Mark L. Weiss
    Abstract:

    Glycoprotein E (gE) is important for full virulence potential of the alphaHerpesviruses in both natural and laboratory hosts. The gE sequence of the neurovirulent Bovine Herpesvirus 5 (BHV-5) was determined and compared with that of the nonneurovirulent BHV-1. Alignment of the predicted amino acid sequences of BHV-1 and BHV-5 gE open reading frames showed that they had 72% identity and 77% similarity. To determine the role of gE in the differential neuropathogenesis of BHV-1 and BHV-5, we have constructed BHV-1 and BHV-5 recombinants: gE-deleted BHV-5 (BHV-5gEDelta), BHV-5 expressing BHV-1 gE (BHV-5gE1), and BHV-1 expressing BHV-5 gE (BHV-1gE5). Neurovirulence properties of these recombinant viruses were analyzed using a rabbit seizure model (S. I. Chowdhury et al., J. Comp. Pathol. 117:295-310, 1997) that distinguished wild-type BHV-1 and -5 based on their differential neuropathogenesis. Intranasal inoculation of BHV-5 gEDelta and BHV-5gE1 produced significantly reduced neurological signs that affected only 10% of the infected rabbits. The recombinant BHV-1gE5 did not invade the central nervous system (CNS). Virus isolation and immunohistochemistry data suggest that these recombinants replicate and spread significantly less efficiently in the brain than BHV-5 gE revertant or wild-type BHV-5, which produced severe neurological signs in 70 to 80% rabbits. Taken together, the results of neurological signs, brain lesions, virus isolation, and immunohistochemistry indicate that BHV-5 gE is important for efficient neural spread and neurovirulence within the CNS and could not be replaced by BHV-1 gE. However, BHV-5 gE is not required for initial viral entry into olfactory pathway.

Rudi Weiblen - One of the best experts on this subject based on the ideXlab platform.

  • Bovine Herpesvirus-5 infection in a rabbit experimental model: Immunohistochemical study of the cellular response in the CNS
    Microbial pathogenesis, 2013
    Co-Authors: Gisele Fabrino Machado, Paulo Michel Roehe, Rudi Weiblen, Fernanda Bernardi, Fernando Y. M. Hosomi, Juliana Regina Peiró, Antonio Carlos Alessi, Guilherme D. Melo, Adriano Tony Ramos, Paulo César Maiorka
    Abstract:

    Universidade Estadual Paulista (UNESP) Faculdade de Medicina Veterinaria Departamento de Clinica, Cirurgia e Reproducao Animal, Aracatuba, SP

  • Recombinante do herpesvírus bovino tipo 5 com deleção na timidina quinase estabelece infecção latente porém reativa ineficientemente em ovinos
    Pesquisa Veterinária Brasileira, 2013
    Co-Authors: Gustavo Cauduro Cadore, Rudi Weiblen, M. Weiss, D Anziliero, M C S Brum, Eduardo Furtado Flores
    Abstract:

    The ability of thymidine kinase (tk)-deleted recombinant Bovine Herpesvirus 5 (BoHV-5tkΔ) to establish and reactivate latent infection was investigated in lambs. During acute infection, the recombinant virus replicated moderately in the nasal mucosa, yet to lower titers than the parental strain. At day 40 post-infection (pi), latent viral DNA was detected in trigeminal ganglia (TG) of all lambs in both groups. However, the amount of recombinant viral DNA in TGs was lower (9.7-fold less) than that of the parental virus as determined by quantitative real time PCR. Thus, tk deletion had no apparent effect on the frequency of latent infection but reduced colonization of TG. Upon dexamethasone (Dx) administration at day 40 pi, lambs inoculated with parental virus shed infectious virus in nasal secretions, contrasting with lack of infectivity in secretions of lambs inoculated with the recombinant virus. Nevertheless, some nasal swabs from the recombinant virus group were positive for viral DNA by PCR, indicating low levels of reactivation. Thus, BoHV-5 TK activity is not required for establishment of latency, but seems critical for efficient virus reactivation upon Dx treatment.

  • Mapeamento dos sítios de latência e reativação pelo herpesvírus bovino tipo 5 (BoHV-5) e por mutante deletado no gene da timidina quinase em ovinos
    Pesquisa Veterinária Brasileira, 2013
    Co-Authors: Gustavo Cauduro Cadore, M C S Brum, Rudi Weiblen, Gian Marcon, Eduardo Furtado Flores
    Abstract:

    A thymidine kinase (tk)-deleted Bovine Herpesvirus 5 (BoHV-5tkΔ) was previously shown to establish latent infection and reactivate - even poorly - in a sheep model (Cadore et al. 2013). As TK-negative alphaHerpesviruses are unlike to reactivate in neural tissue, this study investigated the sites of latency and reactivation by this recombinant in lambs. For this, groups of lambs were inoculated intranasally with the parental BoHV-5 strain (SV-507/99) or with the recombinant BoHV-5tkΔ. During latent infection (40 days post-inoculation, pi), the distribution of recombinant virus DNA in neural and non-neural tissues was similar to that of the parental virus. Parental and recombinant virus DNA was consistently detected by PCR in trigeminal ganglia (TGs); frequently in palatine and pharyngeal tonsils and, less frequently in the retropharyngeal lymph nodes. In addition, latent DNA of both viruses was detected in several areas of the brain. After dexamethasone (Dx) administration (day 40pi), the recombinant virus was barely detected in nasal secretions contrasting with marked shedding of the parental virus. In tissues of lambs euthanized at day 3 post-Dx treatment (pDx), reverse-transcription-PCR (RT-PCR) for a late viral mRNA (glycoprotein D gene) demonstrated reactivation of parental virus in neural (TGs) and lymphoid tissues (tonsils, lymph node). In contrast, recombinant virus mRNA was detected only in lymphoid tissues. These results demonstrate that BoHV-5 and the recombinant BoHV-5tkΔ do establish latent infection in neural and non-neural sites. Reactivation of the recombinant BoHV-5tkΔ, however, appeared to occur only in non-neural sites. In anyway, the ability of a tk-deleted strain to reactivate latent infection deserves attention in the context of vaccine safety.

  • A recombinant Bovine Herpesvirus 5 defective in thymidine kinase and glycoprotein E is immunogenic for calves and confers protection upon homologous challenge and BoHV-1 challenge.
    Veterinary Microbiology, 2011
    Co-Authors: D Anziliero, Rudi Weiblen, C M B Santos, M C S Brum, S I Chowdhury, Eduardo Furtado Flores
    Abstract:

    Abstract A recombinant Bovine Herpesvirus 5 lacking thymidine kinase and glycoprotein E genes (BoHV-5gEΔTKΔ) was evaluated as a live experimental vaccine. In a first experiment, ten-months-old calves were vaccinated intramuscularly ( n  = 9) or remained as controls ( n  = 8) and 42 days later were challenged with BoHV-5 or BoHV-1 intranasally. The four control calves challenged with BoHV-5 developed severe depression and neurological signs and were euthanized in extremis at days 13 and 14 pos-infection (pi); the five vaccinated animals challenged with BoHV-5 remained healthy. The titers of virus shedding were reduced ( p p n  = 15) or kept as controls ( n  = 5) and subsequently challenged with a BoHV-1 isolate. Control calves developed moderate to severe rhinitis and respiratory distress; two were euthanized in extremis at days 5 and 9 pi, respectively. In contrast, vaccinated animals were protected from challenge and only a few developed mild and transient nasal signs. The duration and titers of virus shedding after challenge were reduced ( p

  • A thymidine kinase-negative Bovine Herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection
    Pesquisa Veterinária Brasileira, 2011
    Co-Authors: Sara C. Da Silva, Rudi Weiblen, Mário Celso Speroto Brum, Stephan A. M Oliveira, Eduardo Furtado Flores
    Abstract:

    Mutant viral strains deleted in non-essential genes represent useful tools to study the function of specific gene products in the biology of the virus. We herein describe an investigation on the phenotype of a Bovine Herpesvirus 5 (BoHV-5) recombinant deleted in the gene encoding the enzyme thymidine kinase (TK) in rabbits, with special emphasis to neuroinvasiveness and the ability to establish and reactivate latent infection. Rabbits inoculated with the parental virus (SV-507/99) (n=18) at a low titer (105.5TCID50) shed virus in nasal secretions in titers up to 104.5TCID50 for up to 12 days (average: 9.8 days [5-12]) and 5/ 16 developed neurological disease and were euthanized in extremis. Rabbits inoculated with the recombinant BoHV-5TKΔ at a high dose (107.1TCID50) also shed virus in nasal secretions, yet to lower titers (maximum: 102.3TCID50) and for a shorter period (average: 6.6 days [2-11]) and remained healthy. PCR examination of brain sections of inoculated rabbits at day 6 post-infection (pi) revealed a widespread distribution of the parental virus, whereas DNA of the recombinant BoHV-5TKΔ-was detected only in the trigeminal ganglia [TG] and olfactory bulbs [OB]. Nevertheless, during latent infection (52pi), DNA of the recombinant virus was detected in the TGs, OBs and also in other areas of the brain, demonstrating the ability of the virus to invade the brain. Dexamethasone (Dx) administration at day 65 pi was followed by virus reactivation and shedding by 5/8 rabbits inoculated with the parental strain (mean duration of 4.2 days [1 - 9]) and by none of seven rabbits inoculated with the recombinant virus. Again, PCR examination at day 30 post-Dx treatment revealed the presence of latent DNA in the TGs, OBs and in other areas of the brain of both groups. Taken together, these results confirm that the recombinant BoHV-5TKΔ is highly attenuated for rabbits. It shows a reduced ability to replicate in the nose but retains the ability to invade the brain and to establish latent infection. Additional studies are underway to determine the biological and molecular mechanisms underlying the inability of BoHV-5TKΔ to reactivate from latency.