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1.
Microb Pathog ; 169: 105645, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35716923

RESUMO

Bovine gammaherpesvirus type 4 (BoHV-4) shows tropism for the endometrium, in which it causes the death of epithelial and stroma cells. Despite having anti-apoptotic genes in its genome, experiments based on immortalized cell lines have shown that BoHV-4 induces cell death by apoptosis. In the present study, we evaluated BoHV-4 replication, pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) mitochondrial genes expression and chromatin condensation in bovine endometrium primary culture cells (BEC) and in the Madin Darby bovine kidney (MDBK) cell line. Results showed that BoHV-4 has a preference for replication in BEC cells over the MDBK cell line, demonstrated by the high viral titer that is consistent with the tropism of the virus. In BEC cells, chromatin condensation was consistent with the values of viral kinetics at the late stage of infection, accompanied with a balance in the mRNA levels of apoptotic mitochondrial proteins. As a consequence, in those cells viral transmission would be enhanced by inhibiting apoptosis in the early stage of virus proliferation, allowing the complete production of viral progeny, and then, the induction of apoptosis in late stages would allow neighboring cells infection. In MDBK cells replication kinetics was coincident with the up-regulation of Bcl-2, which suggests that the productive infection in MDBK is associated with a lytic phase of the virus or another cell death pathway (probably autophagy mechanism) at the late stage of infection. The results agree with the study of nuclear morphology, where a constant chromatin condensation was observed over time. It is clear that the documented BoHV-4 apoptotic responses observed in the cell lines studied above are not valid in cells from primary cultures. The data presented in this study suggest that BoHV-4 could induce apoptosis in BEC cells without a leading role of the mitochondria pathway. Further studies will be necessary to characterize in detail the programmed cell death pathways involved in BoHV-4 infection in the primary cell cultures evaluated.


Assuntos
Herpesvirus Bovino 4 , Animais , Apoptose , Bovinos , Linhagem Celular , Cromatina , Feminino , Herpesvirus Bovino 4/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Replicação Viral
2.
Arch Virol ; 167(8): 1659-1668, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35708765

RESUMO

Bovine gammaherpesvirus 4 (BoHV-4) is ubiquitous in cattle worldwide, and it has been detected in animals exhibiting broad clinical presentations. The virus has been detected in the United States since the 1970s; however, its clinical relevance remains unknown. Here, we determined the complete genome sequences of two contemporary BoHV-4 isolates obtained from respiratory (SD16-38) or reproductive (SD16-49) tract specimens and assessed clinical, virological, and pathological outcomes upon intranasal (IN) inoculation of calves with the respiratory BoHV-4 isolate SD16-38. A slight and transient increase in body temperature was observed in BoHV-4-inoculated calves. Additionally, transient viremia and virus shedding in nasal secretions were observed in all inoculated calves. BoHV-4 DNA was detected by nested PCR in the tonsil and regional lymph nodes (LNs) of calves euthanized on day 5 post-inoculation (pi) and in the lungs of calves euthanized on day 10 pi. Calves euthanized on day 35 pi harbored BoHV-4 DNA in the respiratory tract (turbinates, trachea, lungs), regional lymphoid tissues, and trigeminal ganglia. Interestingly, in situ hybridization revealed the presence of BoHV-4 DNA in nerve bundles surrounding the trigeminal ganglia and retropharyngeal lymph nodes (day 35 pi). No histological changes were observed in the respiratory tract (turbinate, trachea, and lung), lymphoid tissues (tonsil, LNs, thymus, and spleen), or central nervous tissues (olfactory bulb and trigeminal ganglia) sampled throughout the animal studies (days 5, 10, and 35 pi). This study contributes to the understanding of the infection dynamics and tissue distribution of BoHV-4 following IN infection in calves. These results suggest that BoHV-4 SD16-38 used in our study has low pathogenicity in calves upon intranasal inoculation.


Assuntos
Doenças dos Bovinos , Infecções por Herpesviridae , Herpesvirus Bovino 1 , Herpesvirus Bovino 4 , Animais , Anticorpos Antivirais , Bovinos , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/genética , Eliminação de Partículas Virais
3.
Front Immunol ; 12: 705539, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34594325

RESUMO

The Morbillivirus peste des petits ruminants virus (PPRV) is the causal agent of a highly contagious disease that mostly affects sheep and goats and produces considerable losses in developing countries. Current PPRV control strategies rely on live-attenuated vaccines, which are not ideal, as they cannot differentiate infected from vaccinated animals (DIVA). Recombinant vector-based vaccines expressing viral subunits can provide an alternative to conventional vaccines, as they can be easily paired with DIVA diagnostic tools. In the present work, we used the bovine herpesvirus-4-based vector (BoHV-4-A) to deliver PPRV hemagglutinin H antigen (BoHV-4-A-PPRV-H-ΔTK). Vaccination with BoHV-4-A-PPRV-H-ΔTK protected sheep from virulent PPRV challenge and prevented virus shedding. Protection correlated with anti-PPRV IgGs, neutralizing antibodies and IFN-γ-producing cells induced by the vaccine. Detection of antibodies exclusively against H-PPRV in animal sera and not against other PPRV viral proteins such as F or N could serve as a DIVA diagnostic test when using BoHV-4-A-PPRV-H-ΔTK as vaccine. Our data indicate that BoHV-4-A-PPRV-H-ΔTK could be a promising new approach for PPRV eradication programs.


Assuntos
Vetores Genéticos , Herpesvirus Bovino 4 , Peste dos Pequenos Ruminantes/prevenção & controle , Vírus da Peste dos Pequenos Ruminantes , Doenças dos Ovinos/imunologia , Ovinos/imunologia , Proteínas Virais , Vacinas Virais , Animais , Chlorocebus aethiops , Cães , Vetores Genéticos/genética , Vetores Genéticos/imunologia , Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/imunologia , Peste dos Pequenos Ruminantes/genética , Peste dos Pequenos Ruminantes/imunologia , Vírus da Peste dos Pequenos Ruminantes/genética , Vírus da Peste dos Pequenos Ruminantes/imunologia , Ovinos/virologia , Doenças dos Ovinos/virologia , Células Vero , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/genética , Vacinas Virais/imunologia
4.
Vet Ital ; 57(1): 49-59, 2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34313098

RESUMO

Bovine herpesvirus type 4 (BoHV­4) is a common virus in the world that is detected in clinically ill or in apparently healthy cattle. This study provides a molecular characterization of BoHV­4 strains from 24 cattle some showing respiratory and/or reproductive problems and some without any apparent clinical sign. This study also reported the growth properties of five BoHV­4 field isolates. The 24 sampled cattle came from 13 different herds in 10 provinces collected between 2007 and 2018. Phylogenetic analysis using partially amplified nucleotide sequences of ORF8 genes coding glycoprotein B (n = 24) and ORF3 genes coding thymidine kinase (n = 9), demonstrated genetic variability among the BoHV­4 strains analysed. The partial gB gene sequences clustered in three different genotypes (genotype I, II and III) were located within the genotype I cluster, such as Movar strain. The analysis of the five BoHV­4 strains isolated from vaginal swabs (n = 2), nasal swab (n = 1), and brain samples (n = 2) revealed no significant differences in their growth properties in MDBK cell culture.


Assuntos
Doenças dos Bovinos/epidemiologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/isolamento & purificação , Criação de Animais Domésticos , Animais , Bovinos , Doenças dos Bovinos/virologia , Infecções por Herpesviridae/epidemiologia , Herpesvirus Bovino 4/genética , Filogenia , Turquia/epidemiologia
5.
Braz J Microbiol ; 52(3): 1119-1133, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34255309

RESUMO

In recent years, Bovine herpesvirus 4 (BoHV-4) has emerged as an attractive gene delivery viral vector, mainly for vaccination purposes in the veterinary field. In the present study, a new infectious clone of the BoHV-4 genome carrying a bacterial artificial chromosome vector (BoHV-4-BAC) was developed by homologous recombination in mammalian cell culture and bacterial systems, and exploited to express a truncated form of glycoprotein D (tgD) of Bovine herpesvirus 1 (BoHV-1) (BoHV-4-tgD∆TK) as a vaccine candidate. This construct's immunogenicity was compared to a DNA vector expressing the same antigen (pC-tgD) in a BALB/c mouse model. After the mice were immunized, total and specific antibody responses, cytokine responses, total splenocyte cells proliferation/cytotoxicity, and virus neutralization assays were conducted to analyze the immune response elicited by both constructs. Mice from both vaccine groups developed significant humoral and cellular immune responses after a booster dose regime was conducted on day 28 post-injection. In almost all immunological assays, BoHV-4-tgDΔTK induced as high an immune response as pC-tgD. In both vaccine constructs, neutralizing antibodies were a significant determining factor in protection against BoHV-1, even after the first injection. We conclude that a BoHV-4-based viral vector offers an effective immunization strategy as an alternative to DNA-based immunization platforms, at least to combat BoHV-1.


Assuntos
Herpesvirus Bovino 1 , Herpesvirus Bovino 4 , Proteínas Virais/imunologia , Animais , Anticorpos Neutralizantes , Anticorpos Antivirais , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/imunologia , Herpesvirus Bovino 4/genética , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Virais/genética
6.
Arch Virol ; 166(2): 619-626, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33410994

RESUMO

Bovine herpesvirus 4 (BoHV-4) is one of the most important of the known viral respiratory and reproductive pathogens of both young and adult cattle. However, BoHV-4 has not been isolated or detected in mainland China prior to this study. In 2019, BoHV-4 strain 512 was isolated from cattle in Heilongjiang Province, China, using MDBK cells, and characterized by PCR, nucleotide sequence analysis, and transmission electron microscopy. Two other unknown herpesvirus strains, BL6010 and J4034, which were isolated from cattle in 2009 in China and stored at -70℃, were also propagated in MDBK cells and identified as BoHV-4 by PCR. Phylogenetic analysis based on partial nucleotide sequences of the thymidine kinase (TK) gene and glycoprotein B (gB) gene for the three isolates indicated that these three Chinese strains belong to BoHV-4 genotype 1. A preliminary virus neutralization test revealed that 64% of the 70 bovine sera (45/70) collected from Inner Mongolia Autonomous Region, China, had anti-BoHV-4 antibodies and that natural BoHV-4 infection occurred in cattle in China. Here, we report for the first time the isolation and molecular characterization of BoHV-4 from cattle in mainland China.


Assuntos
Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/isolamento & purificação , Animais , Sequência de Bases/genética , Bovinos , Doenças dos Bovinos/virologia , China , DNA Viral/genética , Infecções por Herpesviridae/virologia , Timidina Quinase/genética , Proteínas Virais/genética
7.
Arch Virol ; 166(2): 535-544, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33403475

RESUMO

In vitro cell cultures are widely used models for dissecting cellular and molecular mechanisms that lead to certain physiological conditions and diseases. The pathogenesis of BoHV-4 in the bovine reproductive tract has been studied by conducting tests on primary cultures. However, many questions remain to be answered about the role of BoHV-4 in endometrial cells. The aim of this study was to compare the replication and gene expression of BoHV-4 in cell lines and bovine reproductive tract primary cells as an in vitro model for the study of this virus. We demonstrated that BoHV-4 strains differ in their in vitro growth kinetics and gene expression but have the same cell type preference. Our results demonstrate that BoHV-4 replicates preferentially in bovine endometrial cells (BEC). However, its replication capacity extends to various cell types, since all cells that were tested were permissive to BoHV-4 infection. The highest virus titers were obtained in BEC cells. Nevertheless, virus replication efficiency could not be fully predicted from the mRNA expression profiles. This implies that there are multiple cell-type-dependent factors and strain properties that determine the level of BoHV-4 replication. The results of this study provide relevant information about the in vitro behavior of two field isolates of BoHV-4 in different cell cultures. These findings may be useful for the design of future in vitro experiments to obtain reliable results not only about the pathogenic role of BoHV-4 in the bovine female reproductive tract but also in the development of efficient antiviral strategies.


Assuntos
Expressão Gênica/genética , Herpesvirus Bovino 4/genética , Replicação Viral/genética , Animais , Bovinos , Doenças dos Bovinos/virologia , Linhagem Celular , Endométrio/virologia , Feminino , Infecções por Herpesviridae/virologia , Humanos , RNA Mensageiro/genética , Carga Viral/genética
8.
Virus Genes ; 57(1): 31-39, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33104955

RESUMO

Oncolytic viruses have been extensively used in cancer treatment due to their tropism, selective replication only in tumor cells, and possible synergic interaction with other therapeutics. Different researchers have demonstrated that bovine herpesvirus 4 (BoHV-4), a member of the gammaherpesviridae family, has oncolytic potential in some human-origin cancer cell lines like glioma through the selective replication strategy. Using four apoptosis detection methods, namely MTT, LDH, TUNEL, and Annexin V assays, we evaluated the apoptotic effect of BoHV-4 Movar33/63 reference strain along with a recombinant BoHV-4 expressing EGFP in U87 MG cells (human glioblastoma cell line), MDA MB-231 (human breast cancer cell line), and MCF10a (non-tumorigenic human mammary epithelial cell line). Our findings indicate that this virus can replicate and induce apoptosis in these cell lines and hinder in vitro proliferation in a dose-dependent manner. In conclusion, BoHV-4 has in vitro potential as a novel oncolytic virus in human cancer therapy. However, its replication potential in the MCF10a cells as a non-tumorigenic human mammary epithelial cell line is a concern in using this virus in cancer therapy, at least against human mammary tumors. Further studies must therefore be conducted to examine the specific apoptotic pathways induced by this virus to move on to further experiments.


Assuntos
Neoplasias da Mama/terapia , Glioblastoma/terapia , Herpesvirus Bovino 4/fisiologia , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/fisiologia , Replicação Viral , Apoptose , Linhagem Celular Tumoral , Humanos
9.
Vet Microbiol ; 251: 108888, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33120087

RESUMO

Bovine gammaherpesvirus 4 (BoHV-4) has controversially been related with cattle reproductive disease. In the present study we analyze the relationship between exposure to BoHV-4 and reproductive performance in dairy cattle from northwestern Spain. A total of 2022 sera from 50 farms were examined to detect anti-BoHV-4 antibodies. Herd and individual reproductive records were collected to analyze association with exposure to BoHV-4. In addition, 52 abortion cases were examined to detect BoHV-4 DNA. An individual seroprevalence of 66.6 % and a herd prevalence of 98 % were found. Exposure to BoHV-4 increased with age, particularly in individuals between 26-36 months old (OR = 2.7; CI 95 %: 1.2-5.0, compared to animals < 26 months). Seroprevalence was not associated with herd fertility and herd abortion rate, but seropositive animals between 26-36 months presented prolonged calving to fertilizing insemination intervals (HR: 1.4; CI 95 %: 1.2-2.0) as well as higher odds of an unsuccessful 1st insemination (OR: 2.5; CI 95 %: 1.2-5.0). In abortion cases, BoHV-4 DNA was found in 12 vaginal swabs from 5 farms but not in any fetal tissue. Our results reveal an endemic, high and widespread exposure to BoHV-4 among dairy cattle from NW Spain with a limited impact in the reproductive performance of herds. The significantly worse reproductive performance of seropositive animals of 26-36 months of age may be the consequence of the establishment of primo-infections when moving heifers to lactation lots. Our findings may be useful to understand the potential population impact of BoHV-4.


Assuntos
Anticorpos Antivirais/sangue , Doenças dos Bovinos/epidemiologia , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/imunologia , Reprodução , Infecções Tumorais por Vírus/veterinária , Animais , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Indústria de Laticínios , Feminino , Infecções por Herpesviridae/imunologia , Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/patogenicidade , Lactação , Gravidez , Prevalência , Estudos Soroepidemiológicos , Espanha/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Infecções Tumorais por Vírus/imunologia
10.
J Hematol Oncol ; 13(1): 89, 2020 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-32653014

RESUMO

BACKGROUND: The oncolytic viruses have shown promising results for the treatment of multiple myeloma. However, the use of human viruses is limited by the patients' antiviral immune response. In this study, we investigated an alternative oncolytic strategy using non-human pathogen viruses as the bovine viral diarrhea virus (BVDV) that were able to interact with CD46. METHODS: We treated several human myeloma cell lines and non-myeloma cell lines with BVDV to evaluate the expression of CD46 and to study the effect on cell viability by flow cytometry. The possible synergistic effect of bortezomib in combination with BVDV was also tested. Moreover, we infected the bone marrow mononuclear cells obtained from myeloma patients and we checked the BVDV effect on different cell populations, defined by CD138, CD14, CD3, CD19, and CD56 expression evaluated by flow cytometry. Finally, the in vivo BVDV effect was tested in NOD-SCID mice injected subcutaneously with myeloma cell lines. RESULTS: Human myeloma cells were selectively sensitive to BVDV treatment with an increase of cell death and, consequently, of apoptotic markers. Consistently, bone marrow mononuclear cells isolated from myeloma patients treated with BVDV, showed a significant selective decrease of the percentage of viable CD138+ cells. Interestingly, bortezomib pre-treatment significantly increased the cytotoxic effect of BVDV in myeloma cell lines with a synergistic effect. Finally, the in vitro data were confirmed in an in vivo myeloma mouse model showing that BVDV treatment significantly reduced the tumoral burden compared to the vehicle. CONCLUSIONS: Overall, our data indicate, for the first time, a direct oncolytic effect of the BVDV in human myeloma cells suggesting its possible use as novel alternative anti-myeloma virotherapy strategy.


Assuntos
Vírus da Diarreia Viral Bovina , Mieloma Múltiplo/terapia , Terapia Viral Oncolítica , Vírus Oncolíticos , Idoso , Idoso de 80 Anos ou mais , Animais , Antígenos CD/análise , Apoptose , Células da Medula Óssea/química , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/virologia , Bortezomib/farmacologia , Linhagem Celular Tumoral , Efeito Citopatogênico Viral , Vírus da Diarreia Viral Bovina/fisiologia , Feminino , Herpesvirus Bovino 4 , Humanos , Masculino , Proteína Cofatora de Membrana/biossíntese , Proteína Cofatora de Membrana/genética , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Pessoa de Meia-Idade , Mieloma Múltiplo/patologia , Vírus Oncolíticos/fisiologia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Leucemia-Linfoma Linfoblástico de Células Precursoras B/terapia , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patologia , Leucemia-Linfoma Linfoblástico de Células T Precursoras/terapia , Organismos Livres de Patógenos Específicos
11.
Microb Pathog ; 144: 104170, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32224211

RESUMO

Some viruses encode inhibitory factors of apoptosis during infection to prolong cell viability and then to achieve a higher production of viral progeny or facilitate persistent infections. There is evidence that some gammaherpesviruses, including BoHV-4, carry genes that can both inhibit or induce apoptosis. BoHV-4 possesses two genes (ORF16 and ORF71) that code for proteins with anti-apoptotic functions, such as v-Bcl2 and v-Flip, respectively. Thus, it is relevant to study BoHV-4 in relation to the modulation of apoptosis in infected cells as a strategy for persistence in the host. The objective of this work was to analyze whether variations in v-Flip and v- Bcl2 of six phylogenetically divergent Argentinean isolates of BoHV-4 can influence the capacity of these strains to induce apoptosis in cell cultures. In this study, variations were mainly detected in the v-Flip gene and protein of the BoHV-4 strains belonging to genotype 3. Thus, it is possible to infer that sequence variations could be associated with some BoHV-4 genotype. Induction of apoptosis was not a significant event for any of the genetically distinct local isolates of BoHV-4 and there was not an evident relationship between the variability of both genes with the apoptotic effect of the phylogenetically distinct strains.


Assuntos
Apoptose/genética , Herpesvirus Bovino 4/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Argentina , Sequência de Bases , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Bovinos , Doenças dos Bovinos/virologia , Linhagem Celular Tumoral , Genótipo , Células HeLa , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/isolamento & purificação , Humanos , Alinhamento de Sequência
12.
Mol Biol Rep ; 47(6): 4905-4909, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32347419

RESUMO

Bovine herpevsirus 4 (BoHV-4) is a gammaherpesvirus that has been associated with different clinical conditions in cattle. In Argentina, BoHV-4 was detected in diverse bovine samples. The aim of this study was to analyze the genetic relationship of 48 field BoHV-4 strains isolated from cattle in Argentina. According to thymidine kinase (tk) gene sequences, BoHV-4 isolates belong to genotypes 1, 2 and 3. Phylogenetic analyses confirmed the presence of the three previously described viral genotypes. However, some of the studied isolates presented conflicting phylogenetic signals between the studied markers. This suggests a complex evolutionary background, that is a history of recombination, incomplete lineage sorting (deep coalescence) or a combination of these, which requires further study. These potential events make difficult the diagnosis of BoHV-4 from clinical samples of cattle and may pose a significant problem for the control of the virus in the herds.


Assuntos
Herpesvirus Bovino 4/genética , Timidina Quinase/genética , Animais , Argentina , Evolução Biológica , Bovinos/virologia , Doenças dos Bovinos/virologia , DNA Viral/genética , Evolução Molecular , Genótipo , Herpesvirus Bovino 4/isolamento & purificação , Herpesvirus Bovino 4/patogenicidade , Filogenia
13.
Onderstepoort J Vet Res ; 87(1): e1-e5, 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32129636

RESUMO

Numerous viruses, including bovine viral diarrhoea virus (BVDV), bovine herpes virus 1 (BoHV-1) and bovine herpes virus 4 (BoHV-4), and other pathogens are the most common causes of reproductive disorders and are responsible for huge economic losses in livestock production. This study investigates the aetiological role of BoHV-4 in fertility problems such as abortions, stillbirth and birth with unviable calves. Retrospective samples from 38 animals, including 17 aborting cows, 17 aborted foetuses, three stillborn calves and one unviable newborn calf were analysed. The BoHV-4 genome was detected in 25 (65.7%) animals by polymerase chain reaction. In 14 of these infected animals, we detected co-infection with BVDV, while the co-presence of BoHV-1 was also detected in one animal. In addition to the high prevalence of BoHV-4 genome in materials related to fertility problems, isolation of BoHV-4 from the brain of one stillborn calf indicated a causal link between BoHV-4 and fertility problems, such as abortion, stillbirths or birth with unviable calves.


Assuntos
Aborto Animal/epidemiologia , Doenças dos Bovinos/epidemiologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/fisiologia , Natimorto/veterinária , Infecções Tumorais por Vírus/veterinária , Animais , Bovinos , Feminino , Infecções por Herpesviridae/epidemiologia , Humanos , Prevalência , Estudos Retrospectivos , Natimorto/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Turquia/epidemiologia
14.
J Vet Sci ; 21(1): e5, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31940684

RESUMO

The major glycoproteins of bovine gammaherpesvirus 4 (BoHV-4) are gB, gH, gM, gL, and gp180 with gB, gH, and gp180 being the most glycosylated. These glycoproteins participate in cell binding while some act as neutralization targets. Glycosylation of these envelope proteins may be involved in virion protection against neutralization by antibodies. In infected cattle, BoHV-4 induces an immune response characterized by low neutralizing antibody levels or an absence of such antibodies. Therefore, virus seroneutralization in vitro cannot always be easily demonstrated. The aim of this study was to evaluate the neutralizing capacity of 2 Argentine BoHV-4 strains and to associate those findings with the gene expression profiles of the major envelope glycoproteins. Expression of genes coding for the envelope glycoproteins occurred earlier in cells infected with isolate 10/154 than in cells infected with strain 07/435, demonstrating a distinct difference between the strains. Differences in serological response can be attributed to differences in the expression of antigenic proteins or to post-translational modifications that mask neutralizing epitopes. Strain 07/435 induced significantly high titers of neutralizing antibodies in several animal species in addition to bovines. The most relevant serological differences were observed in adult animals. This is the first comprehensive analysis of the expression kinetics of genes coding for BoHV-4 glycoproteins in 2 Argentine strains (genotypes 1 and 2). The results further elucidate the BoHV-4 life cycle and may also help determine the genetic variability of the strains circulating in Argentina.


Assuntos
Antígenos Virais/análise , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/genética , Infecções Tumorais por Vírus/veterinária , Proteínas Virais/análise , Animais , Argentina , Bovinos , Doenças dos Bovinos/imunologia , Cervos , Feminino , Doenças das Cabras/imunologia , Doenças das Cabras/virologia , Cabras , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 4/imunologia , Masculino , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/virologia , Transcrição Genética , Infecções Tumorais por Vírus/imunologia , Infecções Tumorais por Vírus/virologia
15.
Arch Virol ; 165(3): 719-723, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31980937

RESUMO

Bovine herpesvirus 4 (BoHV-4) is increasingly believed to be responsible for several disorders of the bovine reproductive tract. The first characterization of BoHV-4 in Argentina was from samples from an aborted fetus. Argentinean isolates are highly diverse and are phylogenetically grouped in three genotypes. In this study, we describe the isolation of BoHV-4 from a bovine fetus with a gestational age of 8 months and without macroscopic lesions. Genetic analyses revealed that the isolated strain belongs to genotype 2. This is the first report on the presence of infectious BoHV-4 in tissues from an aborted bovine fetus.


Assuntos
Feto Abortado/virologia , Aborto Animal/virologia , Doenças dos Bovinos/virologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/isolamento & purificação , Aborto Animal/epidemiologia , Animais , Argentina/epidemiologia , Sequência de Bases , Bovinos , Doenças dos Bovinos/epidemiologia , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , RNA Viral/genética
16.
Vet Microbiol ; 239: 108495, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31767098

RESUMO

Bovine gammaherpesvirus type 4 (BoHV-4) is increasingly related with reproductive disease in cattle, but its epidemiology is not fully understood. We monitored the serological response and shedding of BoHV-4 in a positive dairy cattle farm with metritis. First, we performed an ELISA to detect BoHV-4 antibodies in all the animals (n = 104). Afterwards, ten seronegative heifers introduced in the production lot and sera samples were monthly taken for four months and then 6-10 months after introduction to detect BoHV-4 antibodies by ELISA. Moreover, a vaginal swab was taken after calving to detect BoHV-4 by PCR. Concurrently, a weekly collection of vaginal and nasal swabs and milk was performed during the first month post-partum in multiparous cows with metritis (n = 14), heifers with metritis (n = 4), heifers without metritis but positive to BoHV-4 (ELISA or PCR) (n = 2) and multiparous cows without metritis (n = 3). Seropositivity was higher in older animals and in the production lot. Three heifers which shed BoHV-4 after parturition resulted seronegative at first but eventually seroconverted. In the same vein, most heifers seroconverted after 6-10 months in the production lot (8/10). Multiparous cows shed virus by various routes: 13/14 (93 %) in vaginal secretions, 7/14 (50 %) in nasal exudates and 7/14 (50 %) in milk. However, in the other groups, shedding was only detected in vaginal swabs from the first week post-partum. Our study describes BoHV-4 shedding in field conditions. Seronegative animals may become horizontally infected when moved to a contaminated environment.


Assuntos
Anticorpos Antivirais/sangue , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/fisiologia , Infecções Tumorais por Vírus/veterinária , Eliminação de Partículas Virais , Animais , Bovinos , Feminino , Infecções por Herpesviridae/sangue , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/virologia , Período Pós-Parto , Soroconversão , Infecções Tumorais por Vírus/sangue , Infecções Tumorais por Vírus/imunologia , Infecções Tumorais por Vírus/virologia , Vagina/virologia
17.
PLoS One ; 14(4): e0215605, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31002724

RESUMO

Recombinant herpesvirus vaccine vectors offer distinct advantages in next-generation vaccine development, primarily due to the ability to establish persistent infections to provide sustainable antigen responses in the host. Recombinant bovine herpesvirus-4 (BoHV-4) has been previously shown to elicit protective immunity in model laboratory animal species against a variety of pathogens. For the first time, we describe the induction of antigen-specific immune responses to two delivered antigens in the host species after intranasal nebulization of recombinant BoHV-4 expressing the chimeric peptide containing the bovine viral diarrhea virus (BVDV) glycoprotein E2 and the bovine herpesvirus 1 (BoHV-1) glycoprotein D (BoHV-4-A-CMV-IgK-gE2gD-TM). In this study, four cattle were immunized via intranasal nebulization with the recombinant BoHV-4 construct. Two of the cattle were previously infected with wild-type BoHV-4, and both developed detectable serologic responses to BVDV and BoHV-1. All four immunized cattle developed detectable viral neutralizing antibody responses to BVDV, and one steer developed a transient viral neutralizing response to BoHV-1. Approximately one year after immunization, immunosuppressive doses of the glucocorticoid dexamethasone were administered intravenously to all four cattle. Within two weeks of immunosuppression, all animals developed viral neutralizing antibody responses to BoHV-1, and all animals maintained BVDV viral neutralizing capacity. Overall, nebulization of BoHV-4-A-CMV-IgK-gE2gD-TM persistently infects cattle, is capable of eliciting antigen-specific immunity following immunization, including in the presence of pre-existing BoHV-4 immunity, and recrudescence of the virus boosts the immune response to BoHV-4-vectored antigens. These results indicate that BoHV-4 is a viable and attractive vaccine delivery platform for use in cattle.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , DNA Recombinante/imunologia , Herpesvirus Bovino 4/imunologia , Vacinas Sintéticas/imunologia , Administração Intranasal , Animais , Antígenos Virais/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/prevenção & controle , Doenças dos Bovinos/virologia , DNA Recombinante/genética , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/imunologia , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/genética , Herpesvirus Bovino 1/imunologia , Herpesvirus Bovino 4/genética , Nebulizadores e Vaporizadores , Vacinação/métodos , Vacinação/veterinária , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Virais/administração & dosagem , Vacinas Virais/genética , Vacinas Virais/imunologia
18.
Comp Immunol Microbiol Infect Dis ; 63: 97-103, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30961825

RESUMO

Bovine gammaherpesvirus 4 (BoHV4) is a member of the family Herpesviridae. In Argentina, BoHV4 was isolated and characterized in 2007 from samples of aborted cows. Argentinean isolates are highly divergent and are classified as: Genotype 1(Movar-like), Genotype 2 (DN599-like) and Genotype 3 (a novel group). The aim of this study was to comparatively evaluate the biological characteristics of six Argentinean BoHV4 field isolates in cell lines from different origins. All strains induced productive infection in the cell lines used, with different degrees of permissiveness. A direct relationship among the times of appearance of cytopathic effect, the growth kinetics, the size of the lysis plaques and the virulent-like behaviour in vitro could not be established. However, although slight, there are differences in the biological behaviour of the BoHV4 fields isolates analyzed. This variability is independent of their genetic classification but would be conditioned by the nature of the infected cells.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/fisiologia , Infecções Tumorais por Vírus/veterinária , Replicação Viral/fisiologia , Animais , Argentina , Bovinos , Linhagem Celular , Chlorocebus aethiops , Efeito Citopatogênico Viral , Cães , Células HeLa , Células Hep G2 , Herpesvirus Bovino 4/genética , Herpesvirus Bovino 4/isolamento & purificação , Humanos , Células Madin Darby de Rim Canino , Células Vero
19.
Viruses ; 11(3)2019 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-30857305

RESUMO

Crimean-Congo hemorrhagic fever virus (CCHFV) is the causative agent of a tick-borne infection with a significant mortality rate of up to 40% in endemic areas, with evidence of geographical expansion. Due to a lack of effective therapeutics and control measures, the development of a protective CCHFV vaccine remains a crucial public health task. This paper describes, for the first time, a Bovine herpesvirus type 4 (BoHV-4)-based viral vector (BoHV4-∆TK-CCHFV-N) and its immunogenicity in BALB/c and protection potential in IFNα/ß/γR-/- mice models in comparison with two routinely used vaccine platforms, namely, Adenovirus type 5 and a DNA vector (pCDNA3.1 myc/His A), expressing the same antigen. All vaccine constructs successfully elicited significantly elevated cytokine levels and specific antibody responses in immunized BALB/c and IFNα/ß/γR-/- mice. However, despite highly specific antibody responses in both animal models, the antibodies produced were unable to neutralize the virus in vitro. In the challenge experiment, only the BoHV4-∆TK-CCHFV-N and Ad5-N constructs produced 100% protection against lethal doses of the CCHFV Ank-2 strain in IFNα/ß/γR-/- mice. The delivery platforms could not be compared due to similar protection rates in IFNα/ß/γR-/- mice. However, during the challenge experiment in the T cell and passive antibody transfer assay, BoHV4-∆TK-CCHFV-N was dominant, with a protection rate of 75% compared to others. In conclusion, vector-based CCHFV N protein expression constitutes an effective approach for vaccine development and BoHV-4 emerged as a strong alternative to previously used viral vectors.


Assuntos
Vetores Genéticos , Vírus da Febre Hemorrágica da Crimeia-Congo/imunologia , Febre Hemorrágica da Crimeia/prevenção & controle , Imunização Passiva , Proteínas do Nucleocapsídeo/imunologia , Receptores de Interferon/genética , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Bovinos , Modelos Animais de Doenças , Feminino , Vírus da Febre Hemorrágica da Crimeia-Congo/genética , Febre Hemorrágica da Crimeia/imunologia , Herpesvirus Bovino 4/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Proteínas do Nucleocapsídeo/genética , Vacinação , Vacinas Virais/genética
20.
Vet Res ; 50(1): 11, 2019 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-30736853

RESUMO

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus that is widespread in cattle. However, only a few studies about the pathogenesis of BoHV-4 primary infection have been reported. In the present study, ex vivo models with bovine nasal and tracheal mucosa explants were used to study the cellular BoHV-4-host interactions. Infection was observed in nasal but not in tracheal epithelial cells. To find a possible correlation between the integrity and restricted infection of the respiratory epithelium, both nasal mucosal and tracheal explants were treated with EGTA, a drug that disrupts the intercellular junctions, before inoculation. The infection was analyzed based on the number of plaques, plaque latitude and number of infected single cells, as determined by immunofluorescence. BoHV-4 infection in nasal mucosal explants was enhanced upon opening the tight junctions with EGTA. Infection in tracheal explants was only found after treatment with EGTA. In addition, primary bovine respiratory epithelial cells (BREC) were isolated, grown at the air-liquid interface and infected either at the apical or basolateral side by BoHV-4. The results showed that BoHV-4 preferentially bound to and entered BREC at the basolateral surfaces of both nasal and tracheal epithelial cells. The percentage of BoHV-4 infection was significantly increased both from nasal and tracheal epithelial cells after treatment with EGTA, which indicates that the BoHV-4 receptor is mainly located at the basolateral surface of these cells. Thus, our findings demonstrate that integrity of the respiratory epithelium is crucial in the host's innate defense against primary BoHV-4 infections.


Assuntos
Doenças dos Bovinos/fisiopatologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 4/fisiologia , Infecções Tumorais por Vírus/veterinária , Animais , Bovinos , Doenças dos Bovinos/virologia , Infecções por Herpesviridae/fisiopatologia , Infecções por Herpesviridae/virologia , Mucosa Respiratória/fisiopatologia , Mucosa Respiratória/virologia , Infecções Tumorais por Vírus/fisiopatologia , Infecções Tumorais por Vírus/virologia
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