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1.
Virology ; 519: 53-63, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29653302

RESUMO

Viral infectivity factor (Vif) encoded by lentiviruses is essential for viral replication and escaping from antiviral activity of host defensive factors APOBEC3. Jembrana disease virus (JDV) causes an acute disease syndrome with approximately 20% case fatality rate in Bali cattle. However, the interplay mechanism between JDV Vif and Bos taurus APOBEC3 (btA3) is poorly understood. In this study, we determined that JDV Vif recruits ElonginB, ElonginC(ELOB/C), Cul2 and RBX1 but without the need of CBF-ß to form E3 ubiquitin ligase and induces the degradation of btA3 proteins. Further investigation identified BC-box (T149LQ151) motif required for ELOB/C binding, Cul2 box (Y167xxxxV/X172) and a zinc-binding motif (H95-C113-H115-C133) required for Cul2 binding in JDV Vif. The precise mechanism of JDV Vif overcoming the antiviral activity of btA3 proteins is helpful for the application of the broad spectrum antiviral drug targeting conserved functional domains of various species Vif proteins in the future.


Assuntos
Desaminases APOBEC/metabolismo , Produtos do Gene vif/metabolismo , Lentivirus Bovinos/metabolismo , Proteólise , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Bovinos , Elonguina/metabolismo , Produtos do Gene vif/química , Produtos do Gene vif/genética , Células HEK293 , Humanos , Lentivirus Bovinos/genética , Ligação Proteica , Ubiquitina-Proteína Ligases/química , Ubiquitina-Proteína Ligases/genética
2.
Vet Immunol Immunopathol ; 149(3-4): 167-76, 2012 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-22776774

RESUMO

Jembrana disease virus (JDV) is an unusual bovine lentivirus that causes an acute and sometimes fatal disease after a short incubation period in Bali cattle (Bos javanicus). The pathological changes occur primarily in lymphoid tissues, which feature proliferating lymphoblastoid-like cells predominantly throughout parafollicular (T-cell) areas, and atrophy of follicles (B-cell) areas. Five Bali cattle were experimentally infected with JDV and all developed typical clinical signs of Jembrana disease characterised by a transient febrile response, enlargement of superficial lymph nodes and a significant leukopenia. Flow cytometric analysis of PBMC during the acute (febrile) disease phase showed that the reduced number of lymphocytes was due to a significant decrease in both the proportion and absolute numbers of CD4(+) T cells, but not CD8(+) T-cells or CD21(+) B-cells. At the end of the febrile phase, total numbers of both CD8(+) T-cells and CD21(+) B-cells increased significantly, while CD4(+) T-cell numbers remained below normal values, resulting in a significantly reduced CD4(+):CD8(+) ratio. We speculate that the persistent depletion of CD4(+) T cells following JDV infection, through lack of CD4(+) T cell help to B cells, may explain the lack of production of JDV-specific antibodies for several weeks after recovery despite an increase in CD21(+) B cell numbers. Further, our previous data showing that IgG(+) plasma cells are targets for JDV infection, correlated with our current data demonstrating an increase in CD8(+) T cell numbers, supports the suggestion that anti-viral cytotoxic T cell or other cell-mediated immune responses may be critical in the recovery process, although this remains to be formally demonstrated for JDV.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Doenças dos Bovinos/virologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/imunologia , Subpopulações de Linfócitos/imunologia , Animais , Relação CD4-CD8/veterinária , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD8-Positivos/citologia , Bovinos , Doenças dos Bovinos/imunologia , Feminino , Citometria de Fluxo/veterinária , Imunofenotipagem/veterinária , Indonésia , Interferon gama/sangue , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/imunologia
3.
Virology ; 404(2): 261-8, 2010 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-20570311

RESUMO

In cattle the interaction between the two genetically and antigenically related bovine lentiviruses, the acutely pathogenic Jembrana disease virus (JDV) and the non-pathogenic Bovine immunodeficiency virus (BIV) has not been reported although both JDV and a BIV-like virus have been reported in the Bali cattle (Bos javanicus) population in Indonesia. The outcome of infection of Bali cattle with the R29 strain of BIV prior to superinfection 42 days later with JDV(TAB/87) was determined. All BIV-inoculated cattle were successfully infected and developed an antibody response to the TM and CA proteins. BIV infection did not prevent subsequent infection with JDV or ameliorate the clinical signs of Jembrana disease in the infected cattle. It did, however, modify the dynamics of the JDV infection with an earlier onset and end of the acute disease process, and a reduction in the duration of viremia that exceeded 10(6) genome copies/ml of plasma.


Assuntos
Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/imunologia , Animais , Temperatura Corporal , Bovinos , Ensaio de Imunoadsorção Enzimática , Vírus da Imunodeficiência Bovina/imunologia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Fatores de Tempo , Carga Viral , Viremia
4.
Curr HIV Res ; 8(1): 53-65, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20210780

RESUMO

Jembrana disease virus (JDV) is the most recently discovered member of the lentivirus family and causes an acute clinical disease in Bali cattle with a fatality rate of approximately 15%. It is genetically related to bovine immunodeficiency virus (BIV) to the extent that infections cannot yet be differentially diagnosed using serological assays due to cross-reacting epitopes. Despite their close genetic relationship the pathogenesis of JDV infection in Bali cattle is very different to that of BIV in cattle and is unusual for a member of this virus family. The dynamics of JDV replication and clearance during the acute stage of Jembrana disease, the viral tropism, molecular analysis of the viral genome and mRNA transcripts, and the current status of vaccine development and diagnostic assays are all reviewed to provide a greater understanding of the factors that make JDV such an unusual lentivirus.


Assuntos
Infecções por Lentivirus/veterinária , Lentivirus Bovinos , Animais , Vetores Artrópodes , Búfalos , Bovinos , Indonésia , Infecções por Lentivirus/prevenção & controle , Infecções por Lentivirus/virologia , Lentivirus Bovinos/genética , Lentivirus Bovinos/imunologia , Lentivirus Bovinos/patogenicidade , Filogenia , Carga Viral , Tropismo Viral , Vacinas Virais/uso terapêutico , Vírion/genética
5.
Virology ; 393(2): 221-7, 2009 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-19733380

RESUMO

Jembrana disease virus (JDV) is an unusual bovine lentivirus which causes a non-follicular proliferation of lymphocytes, a transient immunosuppression and a delayed humoral response in infected Bali cattle in Indonesia. A double-immunofluorescent labeling method was developed to identify the subset of mononuclear cells in which the viral capsid protein could be detected. Viral antigen was present in pleomorphic centroblast-like cells which were identified as IgG-containing cells, including plasma cells, in lymphoid tissues. There was no evidence of infection of CD3(+) T-cells or MAC387(+) monocytes in tissues but large vacuolated cells with a macrophage-like morphology in the lung were found to contain viral antigen although they could not be shown conclusively to be infected. The tropism of JDV for mature IgG-containing cells may be relevant to understanding the pathogenesis of Jembrana disease, the delayed antibody responses and the genetic composition of this atypical lentivirus.


Assuntos
Anticorpos Antivirais/imunologia , Doenças dos Bovinos/imunologia , Imunoglobulina G/imunologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/imunologia , Animais , Anticorpos Monoclonais/imunologia , Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Bovinos , Doenças dos Bovinos/virologia , Feminino , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Pulmão/imunologia , Pulmão/virologia , Tecido Linfoide/imunologia , Tecido Linfoide/virologia , Macrófagos/imunologia , Macrófagos/virologia , Monócitos/imunologia , Monócitos/virologia , Plasmócitos/imunologia , Plasmócitos/virologia
6.
J Virol Methods ; 159(1): 81-6, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19442849

RESUMO

A sensitive diagnostic assay for the detection of infections with the bovine lentivirus Jembrana disease virus (JDV) is required in Indonesia to control the spread of Jembrana disease. Immunoassays are used routinely but are compromised by cross-reactive epitopes in the capsid (CA) protein of JDV and the genetically related bovine immunodeficiency virus (BIV). JDV gag-specific primers were tested in a real-time PCR assay to detect proviral DNA in peripheral blood mononuclear cells from 165 cattle from the Tabanan district of Bali. JDV-specific amplicons were detected in 9% of the cattle and only 33% of the real-time PCR positive cattle were seropositive. The delayed seroconversion that occurs after infection with JDV could explain the low concordance between these assays but other factors may be responsible. BIV proviral DNA was not detected in any of the PBMC DNA samples. A high concordance value of 98.6% was found between the JDV plasma-derived antigen Western blot and the JDV p26-his recombinant protein ELISA. Only 21% of the seropositive cattle had detectable levels of proviral DNA suggesting that the proviral load in recovered cattle is low. A combination of real-time PCR and JDV p26-his ELISA is recommended for the detection of infection with JDV in Indonesia.


Assuntos
Doenças dos Bovinos/diagnóstico , Imunoensaio , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Sequência de Bases , Western Blotting , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/imunologia , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática , Indonésia , Infecções por Lentivirus/sangue , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/imunologia , Lentivirus Bovinos/genética , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão/imunologia , Análise de Sequência de DNA
7.
Virology ; 386(2): 317-24, 2009 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-19261319

RESUMO

The efficacy of a tissue-derived vaccine, which is currently used in Indonesia to control the spread of Jembrana disease in Bali cattle, was determined by quantifying the viral load in plasma following experimental infection with Jembrana disease virus. Virus transmission is most likely to occur during the acute phase of infection when viral titers are greater than 10(6) genomes/ml. Vaccinated cattle were found to have a 96% reduction in viral load above this threshold compared to control cattle. This would reduce the chance of virus transmission as the number of days above the threshold in the vaccinated cattle was reduced by 33%. Viral loads at the onset and resolution of fever were significantly lower in the vaccinated cattle and immune function was maintained with the development of antibody responses to Env proteins within 10-24 days post challenge. There was, however, no significant reduction in the duration of the febrile period in vaccinated animals. The duration and severity of clinical parameters were found to be variable within each group of cattle but the quantification of viral load revealed the benefits of vaccinating to reduce the risk of virus transmission as well as to ameliorate disease.


Assuntos
Doenças dos Bovinos/prevenção & controle , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Feminino , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/prevenção & controle , Infecções por Lentivirus/virologia , Modelos Lineares , RNA Viral/análise , Vacinação/veterinária , Carga Viral
8.
Virology ; 386(2): 310-6, 2009 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-19230948

RESUMO

Jembrana disease virus (JDV) is an acute lentiviral infection of Bali cattle in Indonesia. Data generated during a series of cattle infection experiments was examined and significant differences were identified in the mean plasma viral load on the first and second days of the febrile response in cattle infected with JDV(TAB/87) compared to those infected with JDV(PUL/01). The peak and total viral loads >or=10(6) genome copies/ml during the acute stage of the disease were significantly higher in JDV(TAB/87) infected cattle. JDV(PUL/01) infected cattle developed peak rectal temperatures earlier than the JDV(TAB/87) cattle but there were no differences in the duration of the febrile responses observed for the 2 groups of animals. The plasma viremia was above 10(6) genome copies/ml for almost 3 days longer in JDV(TAB/87) compared to JDV(PUL/01) infected cattle. Atypical responses to infection occurred in approximately 15% of experimentally infected animals, characterized by reduced viral loads, lower or absent febrile responses and absence of p26-specific antibody responses. Most of these cattle developed normal Tm-specific antibody responses between 4-12 weeks post-infection.


Assuntos
Doenças dos Bovinos/virologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/fisiologia , Replicação Viral , Animais , Anticorpos Antivirais/imunologia , Temperatura Corporal , Bovinos , Doenças dos Bovinos/imunologia , Feminino , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Lentivirus Bovinos/genética , Lentivirus Bovinos/imunologia , RNA Viral/genética , Carga Viral
10.
Virus Res ; 135(2): 336-9, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18466992

RESUMO

Jembrana disease virus (JDV) is an unusual bovine lentivirus that causes an acute disease syndrome with a 20% case fatality rate after a short incubation period in Bos javanicus (Bali cattle) in Indonesia. Analysis of tat mRNA transcription patterns has identified up to six differently spliced transcripts indicating that, in common with other lentiviruses, JDV uses a complex splicing pattern. RT-PCR analysis of mRNA transcripts produced during the acute phase of infection with JDV(TAB/87) revealed at least 12 differently spliced transcripts involving 9 different splice sites. A single unspliced gag/pol transcript, singly spliced vif and tmx specific transcripts and alternatively spliced env, tat and rev transcripts were identified. A 67 nucleotide putative non-coding exon was identified that shared the same splice acceptor (SA) as vif and was incorporated into alternative transcripts of tat, rev and env.


Assuntos
Doenças dos Bovinos/virologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/genética , Lentivirus Bovinos/patogenicidade , RNA Mensageiro/metabolismo , Transcrição Genética , Doença Aguda , Animais , Sequência de Bases , Bovinos , Produtos do Gene tat/genética , Produtos do Gene tat/metabolismo , Infecções por Lentivirus/virologia , Lentivirus Bovinos/classificação , Lentivirus Bovinos/metabolismo , Dados de Sequência Molecular , Sítios de Splice de RNA , Splicing de RNA , RNA Mensageiro/química , RNA Mensageiro/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo
11.
J Biol Chem ; 282(39): 28800-28806, 2007 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-17656359

RESUMO

Microtubules are cytoskeletal polymers consisting of tubulin subunits that take part in diverse cell activities. Many viruses hijack cellular motor proteins to move on microtubules toward the cell interior during the entry process and toward the plasma membrane during the egress period. In addition, viruses often remodel microtubules to facilitate the generation of infectious progeny. In this study, we found that the transactivator of transcription protein of Jembrana disease virus (Jtat) bound tubulin and microtubules both in cells and in the purified system. Microtubule co-sedimentation and co-localization assays revealed a robust interaction of Jtat with microtubules. Tubulin turbidity assay further showed that Jtat promoted tubulin polymerization in vitro in a concentration-dependent manner. Moreover, Jtat promoted the partitioning of cellular tubulin toward the polymeric form, increased the level of tubulin acetylation, and significantly enhanced the cold stability of cellular microtubules. In addition, Jtat-mediated disruption of microtubule dynamics induced the release of Bim from microtubules, leading to profound apoptosis. These results not only identify Jtat as an important viral regulator of microtubule dynamics but also indicate that Jtat-induced apoptosis might contribute to Jembrana disease pathogenesis.


Assuntos
Apoptose , Lentivirus Bovinos/fisiologia , Microtúbulos/metabolismo , Transativadores/metabolismo , Tubulina (Proteína)/metabolismo , Internalização do Vírus , Acetilação , Animais , Proteínas Reguladoras de Apoptose/química , Proteínas Reguladoras de Apoptose/metabolismo , Proteína 11 Semelhante a Bcl-2 , Bovinos , Linhagem Celular , Sistema Livre de Células , Humanos , Lentivirus Bovinos/química , Lentivirus Bovinos/patogenicidade , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Microtúbulos/química , Microtúbulos/virologia , Proteínas Proto-Oncogênicas/química , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Transativadores/química , Tubulina (Proteína)/química
12.
Virus Res ; 126(1-2): 233-44, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17433486

RESUMO

Jembrana disease virus (JDV) is a lentivirus associated with an acute disease syndrome with a 20% case fatality rate in Bos javanicus (Bali cattle) in Indonesia, occurring after a short incubation period and with no recurrence of the disease after recovery. Partial regions of gag and pol and the entire env were examined for sequence variation in DNA samples from cases of Jembrana disease obtained from Bali, Sumatra and South Kalimantan in Indonesian Borneo. A high level of nucleotide conservation (97-100%) was observed in gag sequences from samples taken in Bali and Sumatra, indicating that the source of JDV in Sumatra was most likely to have originated from Bali. The pol sequences and, unexpectedly, the env sequences from Bali samples were also well conserved with low nucleotide (96-99%) and amino acid substitutions (95-99%). However, the sample from South Kalimantan (JDV(KAL/01)) contained more divergent sequences, particularly in env (88% identity). Phylogenetic analysis revealed that the JDV(KAL/01)env sequences clustered with the sequence from the Pulukan sample (Bali) from 2001. JDV appears to be remarkably stable genetically and has undergone minor genetic changes over a period of nearly 20 years in Bali despite becoming endemic in the cattle population of the island.


Assuntos
Doenças dos Bovinos/virologia , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Primers do DNA/genética , DNA Viral/genética , Evolução Molecular , Genes env , Genes gag , Genes pol , Instabilidade Genômica , Indonésia , Infecções por Lentivirus/virologia , Lentivirus Bovinos/classificação , Lentivirus Bovinos/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico
13.
Virus Res ; 121(2): 122-33, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16870296

RESUMO

Jembrana disease virus (JDV) is a lentivirus highly related to the bovine immunodeficiency virus (BIV). It causes an acute disease with high mortality rate within 1-2 weeks. JDV encodes the most potent Tat (JTat) of any of the lentiviruses. JTat can transactivate all LTRs and functionally substitute for HIV Tat in the viral genome and may function as a pivotal regulator in the acute pathogenesis of JDV. The goal of this paper is to study JTat internalization by cells, the mechanisms involved in internalization, and the effect of JTat on neighbouring cells. By quantification and fluorescence microscopy, we found that the internalization of extracellular EGFP-JTat fusion protein was both time and dose-dependent, but endocytosis and energy independent. We identified that arginines which were responsible for the internalization. Internalized JTat was distributed in both the nucleus and the cytoplasm, could transactivate JDV LTR and modulate cellular gene expression. Based on our findings, we propose that secretion and internalization of JTat may be a way for JDV to influence neighbouring cells and make the cellular environment more amenable to viral infection.


Assuntos
Produtos do Gene tat/fisiologia , Infecções por Lentivirus/virologia , Lentivirus Bovinos/fisiologia , Sequência de Aminoácidos , Animais , Arginina/fisiologia , Bovinos , Núcleo Celular/metabolismo , Células Cultivadas , Citoplasma/metabolismo , Endocitose , Corantes Fluorescentes , Produtos do Gene tat/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Lentivirus Bovinos/metabolismo , Dados de Sequência Molecular , Transporte Proteico , Proteínas/genética , Proteínas/metabolismo , Sequências Repetidas Terminais/fisiologia , Ativação Transcricional
14.
J Clin Microbiol ; 43(11): 5574-80, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16272489

RESUMO

Jembrana disease virus (JDV) is an acutely pathogenic lentivirus that affects Bali cattle in Indonesia. The inability to propagate the virus in vitro has made it difficult to quantitate JDV and determine the kinetics of virus replication during the acute phase of the disease process. We report for the first time two techniques that enable quantification of the virus and the use of these techniques to quantify the virus load during the acute phase of the disease process. A one-step JDV gag [corrected] TaqMan real-time reverse transcription-PCR (RT-PCR) assay was developed for the detection and quantification of JDV RNA in plasma. The limit of detection was 9.8 x 10(2) JDV viral RNA copies over 35 cycles, equivalent to 4.2 x 10(4) JDV genome copies/ml, and a peak virus load of 1.6 x 10(12) during the acute febrile period. An antigen capture enzyme-linked immunosorbent assay (ELISA) was also developed to quantify the levels of JDV capsid (JDVp26) over a linear range of 10 to 200 ng/ml. Viral RNA and JDVp26 levels were correlated in 48 plasma samples obtained from experimentally infected cattle. A significant positive correlation (R = 0.860 and r(2) = 0.740) was observed between the two techniques within the range of their detection limits. The relatively insensitive capture ELISA provides an economical and feasible method for monitoring of virus in the absence of more sensitive techniques.


Assuntos
Doenças dos Bovinos/virologia , Ensaio de Imunoadsorção Enzimática , Infecções por Lentivirus/veterinária , Lentivirus Bovinos/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Carga Viral/veterinária , Animais , Anticorpos Antivirais , Antígenos Virais/sangue , Antígenos Virais/imunologia , Proteínas do Capsídeo/sangue , Proteínas do Capsídeo/imunologia , Bovinos , Doenças dos Bovinos/diagnóstico , Modelos Animais de Doenças , Progressão da Doença , Ensaio de Imunoadsorção Enzimática/métodos , Estudos de Avaliação como Assunto , Febre/patologia , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/virologia , Lentivirus Bovinos/genética , Lentivirus Bovinos/imunologia , RNA Viral/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade , Taq Polimerase
15.
J Virol Methods ; 124(1-2): 135-42, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15664061

RESUMO

In Indonesia, it is suspected that there are two bovine lentiviruses circulating in the cattle population: a pathogenic Jembrana disease virus (JDV), and a nonpathogenic bovine immunodeficiency-like virus (BIV). Both viruses cross-react antigenically and cannot be differentiated by current serological tests using JDV antigens. To identify possible type-specific epitopes, a series of recombinant protein constructs including the matrix, capsid and nucleocapsid proteins were produced from JDV gag and the expressed proteins were tested by Western blot using JDV and BIV hyperimmune sera. JDV matrix and truncated capsid proteins were recognised by both JDV and BIV hyperimmune sera indicating that there were multiple cross-reactive epitopes present in JDV gag. At least three epitopic regions were identified in these constructs, including the major homology region, by monoclonal antibody binding studies. JDV nucleocapsid recombinant protein was not recognised by either JDV or BIV hyperimmune sera and none of the recombinant gag proteins were able to differentiate between JDV positive sera from Jembrana disease endemic and Jembrana disease-free areas. Additionally, a 40 amino acid recombinant subunit protein encompassing the region recently found to contain an epitope unique to BIV [Zheng, L., Zhang, S., Wood, C., Kapil, S., Wilcox, G.E., Loughin, T.A., Minocha, H.C., 2001. Differentiation of two bovine lentiviruses by a monoclonal antibody on the basis of epitope specificity. Clin. Diagn. Lab. Immunol. 8, 283-287] was tested but was not recognised by either JDV positive sera from Jembrana disease-endemic or Jembrana disease-free areas.


Assuntos
Produtos do Gene gag/imunologia , Vírus da Imunodeficiência Bovina/imunologia , Lentivirus Bovinos/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Bovinos , Reações Cruzadas , Mapeamento de Epitopos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/imunologia
16.
Comp Immunol Microbiol Infect Dis ; 26(2): 89-101, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12493490

RESUMO

Clinical, serological, and pathological abnormalities observed in Holstein cows naturally infected with bovine lentivirus 1 bovine immunodeficiency virus (BIV) and other infections were progressive and most commonly associated with weight loss, lymphoid system deficiency, and behavioral changes. Clinical evidence of meningoencephalitis was dullness, stupor, and occasional head or nose pressing postures. The polymerase chain reactions associated the BIV provirus with the lesions in the central nervous system and lymphoid tissues. Multiple concurrent infections developed in retrovirally infected cows undergoing normal stresses associated with parturition and lactation. A major functional correlate of the lymphoreticular alterations was the development of multiple secondary infections which failed to resolve after appropriate antibacterial therapy. The chronic disease syndrome in dairy cows associated with BIV may be useful as a model system for investigation of the pathogenesis of the nervous system lesions and lymphoid organ changes that occur in humans with lentiviral infection.


Assuntos
Infecções por Lentivirus/veterinária , Lentivirus Bovinos/isolamento & purificação , Animais , Anticorpos Antivirais/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/complicações , Doença das Mucosas por Vírus da Diarreia Viral Bovina/patologia , Encéfalo/patologia , Encéfalo/virologia , Bovinos , DNA Viral/química , DNA Viral/genética , Vírus da Diarreia Viral Bovina/isolamento & purificação , Feminino , Histocitoquímica/veterinária , Infecções por Lentivirus/sangue , Infecções por Lentivirus/complicações , Infecções por Lentivirus/patologia , Lentivirus Bovinos/genética , Tecido Linfoide/patologia , Tecido Linfoide/virologia , Reação em Cadeia da Polimerase/veterinária
17.
Clin Diagn Lab Immunol ; 9(6): 1277-81, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12414761

RESUMO

Bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV) are closely related bovine lentiviruses that are difficult to distinguish by presently available diagnostic methods. Recently, in our laboratory, a monoclonal antibody (MAb; MAb 10H1) against the BIV Gag protein identified a differential epitope, located at the 6.4-kDa N terminus of a 29-kDa Gag capsid protein, which was absent in JDV. To define the essential amino acids of the epitope, a series of primers within the 163 bp of DNA corresponding to the 6.4-kDa protein were designed. The full-length 163-bp DNA fragment and the smaller DNA fragments with deletions were amplified by PCR and then cloned into pQE32 vectors for protein expression studies. The expressed proteins were analyzed with MAb 10H1 by Western blotting. The differential epitope has been narrowed to a 26-amino-acid region (R121 to R146), which includes 6 residues of p16(MA) (where MA represents the matrix protein) and 20 residues of p2L. A synthetic peptide corresponding to the putative 26-amino-acid epitope blocked MAb 10H1 binding to the expressed peptide. These experiments revealed that the epitope spans the cleavage site between p16(MA) and p2L and presumably will be valuable in distinguishing the two viruses.


Assuntos
Mapeamento de Epitopos , Produtos do Gene gag/imunologia , Vírus da Imunodeficiência Bovina/química , Sequência de Aminoácidos , Animais , Bovinos , Clonagem Molecular , Produtos do Gene gag/química , Produtos do Gene gag/genética , Lentivirus Bovinos/química , Dados de Sequência Molecular
18.
Arch Virol ; 147(3): 643-9, 2002 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11958463

RESUMO

The ability of BIV strain R29 to infect bovine cell lines in the presence or absence of a functional lentiviral Tat protein is described. Jembrana disease virus (JDV) Tat protein was stably expressed in MDBK cells. No viral replication could be detected in this cell line after infection with BIV R29. Transfection of MDBK cells and MDBK Tat expressing cells with BIV R29 proviral DNA established that BIV R29 could not replicate in MDBK cells. Whether viral entry into MDBK cells is also a block to BIV R29 infection of MDBK cells has yet to be established.


Assuntos
Produtos do Gene tat/metabolismo , Vírus da Imunodeficiência Bovina/fisiologia , Replicação Viral , Animais , Bovinos , Linhagem Celular , DNA Viral/fisiologia , Produtos do Gene tat/genética , Vírus da Imunodeficiência Bovina/genética , Vírus da Imunodeficiência Bovina/patogenicidade , Lentivirus Bovinos/genética , Lentivirus Bovinos/metabolismo , Provírus , Transfecção
20.
Vet Microbiol ; 80(4): 313-27, 2001 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-11348768

RESUMO

Antibodies directed against two bovine lentiviruses, Jembrana disease virus (JDV) and bovine immunodeficiency virus (BIV), were detected in Balinese cattle sera using two new enzyme-linked immunosorbent assays (ELISAs) based on the combination of capsid (CA) protein and transmembrane (TM) peptides derived from JDV or BIV sequences. Twenty eight of the 77 sera tested on the JDV ELISA showed anti-JDV antibodies with an unequal distribution of seropositive animals throughout the different districts of Bali. Furthermore, when 17 of the JDV positive sera were tested on Western blot, using the same JDV CA antigen, only 13 were judged positive confirming that the ELISA was a more sensitive technique for the detection of seropositive animals. Finally, 9 of the 49 JDV seronegative animals showed anti-BIV antibodies when tested on BIV-specific ELISA. These two ELISAs appeared to be highly sensitive for the detection of anti-JDV and anti-BIV antibodies. Moreover, for the first time, animals showing antibodies against BIV were identified on the main island of Bali and on the JDV-free Nusa Penida island.


Assuntos
Bovinos/virologia , Ensaio de Imunoadsorção Enzimática/veterinária , Lentivirus Bovinos/isolamento & purificação , Animais , Anticorpos Antivirais/análise , Capsídeo/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Vírus da Imunodeficiência Bovina/isolamento & purificação , Indonésia , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Proteínas da Matriz Viral/imunologia
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