Bovine Infected Cell Protein 0 (bICP0) is an immediate-early protein encoded by Bovine Herpesvirus-1 that modulates host immune response, activates transcription for all viral promoters, and causes ubiquitin-dependent degradation of proteins. Presented herein is a bICP0 protein-protein interaction (PPI) profile, consisting of 98 cellular and 15 viral proteins, generated through co-immunoprecipitation of bICP0 and its binding partners. The PPI profile was analyzed computationally to identify potential sites of interaction with bICP0 and any cellular pathways that may be influenced by bICP0. Some interactors fall in conjunction with bICP0’s known roles during infection, and others are consistent with known associations of bICP0 homologs. However, some proteins in the PPI profile are involved in apoptosis signaling and mRNA spicing—processes both significant during viral infection and novel to the known functions of bICP0 and its homologs. The interaction and co-localization of some of these proteins with bICP0 was further examined.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Bovine herpesvirus type 5 (BHV-5) is a major etiologic agent of meningo-encephalitis in cattle and establishes lifelong latent infection in trigeminal ganglia and also in other areas of the brain. Colonization of deep areas of the brain with latent viral DNA may have important implications on the pathogenesis of BHV-5 neurological infection upon reactivation. In this study, we investigated the distribution of BHV-5 DNA in the brain of experimentally infected rabbits a laboratory model for BHV-5 infection - prior and subsequently to virus reactivation, using a nested PCR for the glycoprotein B gene. Eighteen rabbits inoculated intranasally with a Brazilian BHV-5 isolate were divided in two groups: group A rabbits (n=8) were euthanized 60 days post-inoculation (pi) for tissue collection; group B (n=7) were submitted to dexamethasone administration at day 60pi for reactivation of latent infection and euthanized for tissue collection 60 days later. To compare, we used two groups of BHV-1-infected rabbits (C, n=3 and D, n=3), each group being submitted to one of the above treatments, respectively. In group A rabbits, viral DNA was consistently detected in trigeminal ganglia (8/8), frequently in cerebellum (6/8), anterior cortex, pons medulla (3/8) and only occasionally in thalamus (2/8), ventro-lateral, dorsal and posterior cortices, midbrain (1/8). In rabbits previously submitted to virus reactivation, viral DNA showed a broader distribution, being detected more frequently besides the TG (7/7) - in ventro-lateral (6/7) and posterior cortices (5/7), pons-medulla and thalamus (4/7) and midbrain (3/7). In contrast, rabbits inoculated with BHV-1 harbored latent viral DNA in a few tissues in addition to TG and did not show significant changes in distribution of viral DNA post-reactivation. These results demonstrate that latency by BHV-5 DNA and not BHV-1 DNA may be established in several areas of the brain of experimentally infected rabbits. Further, dexamethasone-induced virus reactivation is followed by a wider distribution of latent viral DNA, probably due to virus dissemination from the original sites. Thus, it is reasonable to speculate that reactivation of latent infection from deep areas of the brain may contribute to the recrudescence of neurological disease frequently observed upon reactivation of latent BHV-5 infection. / O herpesvírus bovino tipo 5 (BHV-5) é um importante agente etiológico de meningoencefalite em bovinos e estabelece infecção latente em seus hospedeiros, principalmente nos gânglios dos nervos sensoriais. No entanto, a colonização de áreas profundas do cérebro com DNA viral pode ter implicações importantes na patogenia da infecção pelo BHV-5 após a reativação da infecção latente. Neste estudo, foi investigada a distribuição do DNA do BHV-5 no cérebro de coelhos infectados experimentalmente, antes e após a reativação da infecção latente, utilizando um nested-PCR para uma seqüência do gene da glicoproteína B. Dezoito coelhos infectados pela via intranasal com um isolado brasileiro de BHV-5 foram divididos em dois grupos: coelhos do grupo A (n=8) foram submetidos a eutanásia 60 dias pós inoculação (pi) para a coleta de tecidos; animais do grupo B (n=7) foram submetidos a administração de dexametasona no dia 60 pi. Para comparação foram utilizados dois grupos de coelhos inoculados com o BHV-1 (C, n=3 e D, n=3), cada grupo sendo submetido a um dos tratamentos acima, respectivamente. Nos animais do grupo A, o DNA viral foi consistentemente detectado no gânglio trigêmeo (8/8), freqüentemente no cerebelo (6/8), com menor freqüência na ponte e córtex anterior (3/8) e ocasionalmente no tálamo (2/8) e córtices ventro-lateral, dorso-lateral e posterior, pedúnculo cerebral e tálamo (1/8). Nos animais previamente submetidos à reativação, a distribuição do DNA viral foi mais ampla, sendo detectado mais consistentemente, além do TG (7/7), nos córtices ventro-lateral (6/7), e posterior (5/7), ponte e tálamo (4/7) e menos freqüentemente no pedúnculo cerebral (3/7). Em contrapartida, os animais inoculados com o BHV-1 apresentaram o DNA viral latente em poucos tecidos além do TG, e não apresentaram alterações importantes na distribuição do DNA viral após a reativação. Esses resultados demonstraram que o DNA latente do BHV-5 - e não o do BHV-1 - pode estar presente em várias áreas do cérebro de coelhos infectados experimentalmente. A reativação induzida por dexametasona resulta na reativação viral e provavelmente na colonização de áreas adicionais no cérebro. Com base nesses resultados pode-se especular que a reativação do DNA viral latente nestas regiões pode contribuir para a recrudescência de doença neurológica observada após a reativação da infecção latente pelo BHV-5.

Made available in DSpace on 2016-04-28T20:18:28Z (GMT). No. of bitstreams: 1 2005- Paula Amorim Schiavo.pdf: 7993139 bytes, checksum: f29a61deafec7746d4a362ff19771732 (MD5) Previous issue date: 2005-03-07 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / BHV-1, the agent of IBR, IPV and IPB, is spread among cattle and buffalo herds, bringing sanitary and economic losses to the livestock industry in Brazil and the world. These diseases can present clinical signs that fake Food-and-Mouth Disease and other vesicular diseases, and differential diagnosis in referenced laboratories is required. The high incidence is a hazardous risk for livestock, as the infection has an insidious character, with a subclinical form, generally unnoticed by the people involved with the cattle maintenance. As the control of the infection is required but there are few reasonable alternatives for the situation within Brazilian herds, we need new strategies for the control of the disease, which will be of major concern for fighting the infection in the world. Vaccines can t protect from infection and confuse diagnostics. Using attenuated vaccines a latent infection can be started. The disease will be the next sanitary barrier for the international trade of animals and products in the European Union. BHV-1 has been eradicated from Austria, Denmark, Finland, Sweden and Switzerland and control programs have started in some other countries. In the present research, we appreciated clinical and epidemiological aspects of the disease, evaluating the BHV-1 presence in Rio de Janeiro state. We have proposed a new method for the diagnostics of infected breeding cows, using the neutralization test in microplates with MDBK monolayers. We have evaluated the efficiency of passive immunization after colostrum ingestion. The effect of Kalanchoe brasiliensis extracts against BHV-1 in MDBK cells was evaluated. The control of the infection with homeopathic and phytotherapy therapeutics is proposed and the publications about BHV-1 vaccination were reviewed. Measures for the control of the infection in cattle herds from Rio de Janeiro State are suggested. / O BHV-1 ? conhecido genericamente como o agente etiol?gico da IBR, da IPV e da IPB, e est? disseminado em rebanhos bovinos e bubalinos do Brasil e do mundo, causando preju?zos econ?micos e sanit?rios ? atividade pecu?ria. Ainda pode apresentar sintomatologia semelhante ? da Febre Aftosa e outras enfermidades vesiculares, de forma que o diagn?stico diferencial de laborat?rio ? exigido. A alta preval?ncia de animais positivos representa um s?rio risco para a atividade pecu?ria, considerando-se a recidividade da infec??o, que apresenta uma forma subcl?nica, facilmente despercebida pelos envolvidos com a bovinocultura. Sendo objeto de controle em todo o mundo, mas com poucas alternativas vi?veis na situa??o atual do rebanho brasileiro, urge a defini??o de novas estrat?gias de controle da infec??o. A vacina??o n?o protege da infec??o, prejudica a identifica??o dos animais portadores e, no caso de vacinas atenuadas, pode estabelecer infec??o latente. A enfermidade ser? a pr?xima barreira sanit?ria ao com?rcio internacional de animais vivos e seus produtos na Uni?o Europ?ia e j? foi erradicada da ?ustria, Dinamarca, Finl?ndia, Su??a e Su?cia, e programas de controle foram iniciados em outros pa?ses europeus. Neste trabalho abordamos aspectos cl?nicos e epidemiol?gicos da doen?a, avaliando a ocorr?ncia da infec??o pelo BHV-1 no Estado do Rio de Janeiro. Tamb?m propomos uma nova metodologia para o diagn?stico de matrizes infectadas pelo BHV-1, usando a t?cnica de neutraliza??o em microplacas com c?lulas MDBK cultivadas em monocamadas; avaliamos a efici?ncia da imuniza??o passiva, a partir da ingest?o do colostro; apresentamos propostas de controle da doen?a com tratamentos homeop?ticos e fitoter?picos; avaliamos o efeito do extrato de Kalanchoe brasiliensis frente ao BHV-1 in vitro; revisamos a literatura referente ? vacina??o contra a doen?a e sugerimos medidas para o controle da doen?a em rebanhos do Estado do Rio de Janeiro.