RESUMO
Thirty-one bovine cutaneous warts were submitted to macroscopic and histological analyses and to molecular analyses to partial amplification and sequencing of the L1 gene of bovine papillomavirus (BPV). Viral types detected were BPV1 (52%), BPV2 (29%), BPV6 (16%) and BPV10 (3%). BPV2 had lower frequency in papilloma in comparison to that in fibropapilloma (p = 0.002).
Assuntos
Papiloma , Papillomaviridae , Infecções por Papillomavirus/veterinária , Verrugas , Animais , Papillomavirus Bovino 1/genética , Papillomavirus Bovino 1/isolamento & purificação , Papillomavirus Bovino 1/patogenicidade , Bovinos , Doenças dos Bovinos/virologia , DNA Viral/genética , Papiloma/patologia , Papiloma/virologia , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Papillomaviridae/patogenicidade , Infecções por Papillomavirus/virologia , Pele/patologia , Pele/virologia , Verrugas/patologia , Verrugas/virologiaRESUMO
Papillomaviruses are among the most widespread animal viruses, with many hosts harbouring multiple virus types. The present study aimed to evaluate the haematological and immunophenotypic profile of cattle infected with bovine papillomavirus (BPV). Blood samples were collected from 10 animals with clinical cutaneous BPV and without clinical papillomatosis (control). Haematological analysis demonstrated a significant reduction in haemoglobin and haematocrit for BPV-infected animals. The results also showed an increase of natural killer cells and a decrease of γδ+ T-cells and the CD4+/CD8+ ratio for the BPV group when compared to the control group. The infection was also found to stimulate a pro-inflammatory profile with the participation of CD8+T cells producing elevated IFN-γ and IL-17. These findings, although preliminary, provide a better understanding of the immune response of cattle infected with BPV.
Assuntos
Papillomavirus Bovino 1/isolamento & purificação , Doenças dos Bovinos/imunologia , Infecções por Papillomavirus/veterinária , Animais , Papillomavirus Bovino 1/patogenicidade , Linfócitos T CD4-Positivos , Linfócitos T CD8-Positivos , Estudos de Casos e Controles , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/virologia , Feminino , Imunofenotipagem/veterinária , Células Matadoras Naturais , Masculino , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/virologiaRESUMO
Bovine papillomavirus (BPV) is an oncogenic virus with mucous and epithelial tropism. Possible productive virus infection in other tissues, such as blood, has been hypothesized. In order to investigate this possibility, three samples of skin papillomas and blood were collected from bovines with BPV infection and five samples of peripheral blood and one sample of normal tissue were collected from a calf without BPV infection. Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood and examined by reverse transcription-polymerase chain reaction, immunofluorescence, in situ hybridization, and electron microscopy. The tissue samples were examined for histopathological and immunohistochemical features. The skin papillomas showed the presence of DNA sequences of BPV-2, BPV-11, and a putative virus type. The blood samples showed DNA sequences of BPV-1, 2, and 4 simultaneously. Immunohistochemistry showed BPV L1 protein in both epithelium and stroma and BPV E2 protein in koilocytes. In situ hybridization confirmed the presence of BPV DNA in PBMCs and immunofluorescence showed nuclear labeling of E2 and L1 BPV proteins in PBMCs. The transcription analysis revealed transcripts of BPV-1 L1, BPV-2 L2, and BPV-4 E7 in blood and papilloma samples of BPV-infected cattle. The comet assay revealed high levels of host cell DNA damage upon BPV infection. Electron microscopy analysis of PBMCs identified the presence of particles in the cytoplasm that are consistent with papillomavirus in size and shape. The productive infection of PBMCs with BPV has been previously discussed and this study provides evidence indicating that PBMCs are a target of BPV.
Assuntos
Papillomavirus Bovino 1/isolamento & purificação , Monócitos/virologia , Papiloma/veterinária , Neoplasias Cutâneas/veterinária , Animais , Papillomavirus Bovino 1/patogenicidade , Bovinos , Epitélio/virologia , Papiloma/sangue , Papiloma/virologia , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/virologiaRESUMO
Bovine papillomavirus (BPV) is considered a useful model to study HPV oncogenic process. BPV interacts with the host chromatin, resulting in DNA damage, which is attributed to E5, E6, and E7 viral oncoproteins activity. However, the oncogenic mechanisms of BPV E6 oncoprotein per se remain unknown. This study aimed to evaluate the mutagenic potential of Bos taurus papillomavirus type 1 (BPV-1) E6 recombinant oncoprotein by the cytokinesis-block micronucleus assay (CBMNA) and comet assay (CA). Peripheral blood samples of five calves were collected. Samples were subjected to molecular diagnosis, which did not reveal presence of BPV sequences. Samples were treated with 1 µg/mL of BPV-1 E6 oncoprotein and 50 µg/mL of cyclophosphamide (positive control). Negative controls were not submitted to any treatment. The samples were submitted to the CBMNA and CA. The results showed that BPV E6 oncoprotein induces clastogenesis per se, which is indicative of genomic instability. These results allowed better understanding the mechanism of cancer promotion associated with the BPV E6 oncoprotein and revealed that this oncoprotein can induce carcinogenesis per se. E6 recombinant oncoprotein has been suggested as a possible vaccine candidate. Results pointed out that BPV E6 recombinant oncoprotein modifications are required to use it as vaccine.
Assuntos
Papillomavirus Bovino 1/genética , Proteínas Oncogênicas Virais/genética , Infecções por Papillomavirus/genética , Proteínas Recombinantes/genética , Animais , Papillomavirus Bovino 1/patogenicidade , Carcinogênese/genética , Bovinos , Linhagem Celular , Ciclofosfamida/administração & dosagem , Instabilidade Genômica/efeitos dos fármacos , Humanos , Proteínas Oncogênicas Virais/administração & dosagem , Papillomaviridae/genética , Papillomaviridae/patogenicidade , Infecções por Papillomavirus/virologia , Proteínas Recombinantes/administração & dosagemRESUMO
Enzootic haematuria and urinary bladder cancer in cattle are associated with feeding on bracken fern and bovine papillomavirus (BPV) infection. An increased rate of chromosomal aberrations in peripheral blood lymphocytes from chronically affected haematuric cows raised in bracken fern pastures has been reported, suggesting the presence of BPV in the peripheral blood of afflicted animals. The purpose of the present investigation was to examine the role of peripheral blood as a potential BPV-transmitting agent and search for clastogenic effects in experimentally infected animals kept on a bracken fern-free diet. Healthy cows were inoculated with blood samples of haematuric animals every two weeks for 18 months. Recipient cows, their offspring, donor animals and a control group were kept on a bracken fern-free diet throughout the experiment. Clinical and molecular analyses for detection of BPV infection were carried out periodically in all groups. Short-term lymphocyte cultures were performed to assess chromosomal aberration levels. The donor cows, the recipient cows and their offspring presented increased levels of chromosomal aberrations. BPV-2 DNA was identified by Southern blotting, PCR and cycle-sequencing of PCR products in peripheral blood of donor and recipient animals and in the progeny of recipient animals. Data support both the concept that BPV can be transmitted through blood and the hypothesis that infection with the virus causes the clastogenic alterations observed in the present experimental model. The presence of BPV-2 DNA and chromosomal alterations in peripheral blood of offspring at the moment of birth is evidence for vertical transmission of BPV.
Assuntos
Papillomavirus Bovino 1/patogenicidade , Doenças dos Bovinos/genética , Doenças dos Bovinos/transmissão , Aberrações Cromossômicas , Infecções por Papillomavirus/veterinária , Infecções Tumorais por Vírus/veterinária , Ração Animal/efeitos adversos , Animais , Transfusão de Sangue , Papillomavirus Bovino 1/genética , Papillomavirus Bovino 1/isolamento & purificação , Bovinos , Doenças dos Bovinos/etiologia , Cocarcinogênese , Primers do DNA/genética , DNA Viral/genética , DNA Viral/isolamento & purificação , Transmissão de Doença Infecciosa , Feminino , Hematúria/genética , Hematúria/veterinária , Transmissão Vertical de Doenças Infecciosas , Modelos Biológicos , Dados de Sequência Molecular , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/transmissão , Reação em Cadeia da Polimerase , Gravidez , Infecções Tumorais por Vírus/genética , Infecções Tumorais por Vírus/transmissão , Neoplasias da Bexiga Urinária/etiologia , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/veterináriaRESUMO
En el presente trabajo se trató de determinar la acción condicionante de la Ciclofosfamida (CF) en la resistencia de los pollos a la acción del virus de la encefalitis equina venezolana (EEV). Se utilizó como inmunosupresor la CF inyectada vía intraperitoneal (ip) en cantidad de 60 mg/Kg de peso y administrada en grupos de pollos a razón de 1,2 y 3 dósis, previamente infectados con virus EEV (cepa V-38, subgrupo 1-A). Se orientó el estudio fundamentalmente hacia los cambios histológicos a nivel del S.N.C.; como también los cambios hemáticos y la respuesta inmunológica en la producción de anticuerpos neutralizantes