Dissertations / Theses on the topic 'Orf virus'
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Gemeniano, Maria Lourdes Charmaine. "Characterization of Orf-A of feline immunodeficiency virus /." For electronic version search Digital dissertations database. Restricted to UC campuses. Access is free to UC campus dissertations, 2002. http://uclibs.org/PID/11984.
Full textWestphal, Dana, and n/a. "Characterisation of a novel inhibitor of apoptosis expressed by Orf virus." University of Otago. Department of Microbiology & Immunology, 2008. http://adt.otago.ac.nz./public/adt-NZDU20080922.162136.
Full textMcKeever, Declan James. "Studies of the immunology and epidemiology of orf." Thesis, University of Edinburgh, 1987. http://hdl.handle.net/1842/30488.
Full textMo, Min, and n/a. "Characterization of an Orf virus RING-H2 protein, B5L : a mimic of cellular anaphase promoting complex subunit 11." University of Otago. Department of Microbiology & Immunology, 2009. http://adt.otago.ac.nz./public/adt-NZDU20090220.085825.
Full textChand, Puran. "Molecular and immunological characterisation of a major envelope protein of capripoxvirus." Thesis, University of Surrey, 1992. http://epubs.surrey.ac.uk/2774/.
Full textDickinson, Victoria Jane. "The cloning and subcellular localisation of maize streak virus ORF V1." Thesis, University of Hull, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321050.
Full textLear, Andrea. "The characterisation of the ovine skin response to orf virus infection." Thesis, University of Edinburgh, 1995. http://hdl.handle.net/1842/20625.
Full textCargnelutti, Juliana Felipetto. "Infecção experimental de coelhos e camundongos com o vírus do ectima contagioso." Universidade Federal de Santa Maria, 2010. http://repositorio.ufsm.br/handle/1/10060.
Full textContagious ecthyma (orf) is a cutaneous disease that affects sheep and goats, and may be occasionally transmitted to humans. The disease is caused by orf virus (ORFV). ORFV infection produces croustous and proliferative lesions, usually on the nostrils and labial commissures of lambs, and also in the udder, teat skin and coronary bands of adults animals. The pathogenesis of ORFV infection is poorly understood and a search for an adequate animal model is required, yet the disease has been already reproduced in sheep, goats and rabbits. This dissertation relates the clinical, virological and pathological aspects of ORFV infection in rabbits and mice experimental inoculated. Ten rabbits, ten mice and two lambs were inoculated intradermally after skin scarification with an hypodermic needle. A viral suspension of ORFV IA-82 strain (108.5TCID50/mL) was inoculated in the internal face of the ear, back skin and labial commissure of rabbits; internal face of the ear of mice. Lambs were inoculated in the labial commissures and in the internal face of hind limbs. All animals were monitored clinically, virologically, and pathologically for 21 days. All rabbits developed clinical signs in the inoculation sites, begining with mild hyperemia that evolved to macules, papules, vesicle, pustules and scabs. Lesions appeared at days 3 and 4 post-inoculation (pi) and lasted to 3 to 10 days. Viral shedding was detected from days 2 to 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusions in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit much milder clinical course was observed in 5 out of 10 inoculated mice. All lambs presented characteristic contagious ecthyma clinical and histopathologycal lesions from days 3 to 18pi, and the virus was recovered from lesions between days 2 and 19pi. At day 28pi, seroneutralization test (SN) was unable to detect neutralizing antibodies in all inoculated animals. These findings show that ORFV replicates and produce local lesions in rabbits and mice. However, rabbits are more susceptible to infection and disease, and may be used as an animal model to study some aspects of ORFV pathogenesis.
O ectima contagioso (ou orf) é uma doença infecto-contagiosa de pele que afeta principalmente ovinos e caprinos, e que ocasionalmente pode acometer o homem. O seu agente etiológico é o vírus da orf (ORFV). O ORFV produz lesões proliferativas, geralmente na comissura labial e no plano naso-labial de cordeiros, e também na pele do úbere, nos tetos e no rodete coronário dos cascos de animais adultos. A patogenia da infecção pelo ORFV é pouco conhecida, embora a doença já tenha sido reproduzida em ovinos, caprinos e coelhos. Essa dissertação relata os achados clínicos, virológicos e histopatológicos da infecção experimental de coelhos e camundongos pelo ORFV. Para isso, coelhos, camundongos e cordeiros foram inoculados pela via intradérmica (ID), após escarificação da pele com agulha hipodérmica. A inoculação dos cordeiros serviu como controle positivo. Uma suspensão viral da cepa IA-82 do ORFV (108,5DICC50/mL) foi inoculada na face interna da orelha, na pele do dorso e na comissura labial dos coelhos; na face interna da orelha dos camundongos; e na comissura labial e face interna do membro pélvico dos cordeiros. Os animais foram monitorados por 21 dias nos aspectos clínicos, virológicos e patológicos. Todos os coelhos inoculados apresentaram lesões semelhantes nos locais de inoculação, iniciando com hiperemia, evoluindo para máculas, pápulas, vesículas, pústulas e crostas. Os sinais surgiram 3 a 4 dias pós inoculação (pi) e duraram por 3 a 10 dias. Excreção viral foi detectada entre os dias 2 e 14pi. A análise histológica das lesões revelou dermatite focal proliferativa, com degeneração balonosa e corpúsculos de inclusão intracitoplasmáticos eosinofílicos nos queratinócitos, semelhante às alterações histológicas observadas nos cordeiros. Lesões similares, mas de menor intensidade foram observadas em 5 de 10 camundongos. Os cordeiros, utilizados como controles positivos, apresentaram lesões clínicas e histopatológicas características de ectima contagioso entre os dias 3 e 18pi, sendo que o vírus foi recuperado das lesões entre os dias 2 e 19dpi. No dia 28pi, pelo teste de soroneutralização (SN), não foram detectados anticorpos neutralizantes no soro dos animais inoculados. Esses resultados demonstram que a inoculação de ORFV resulta em replicação viral e produção de lesões em coelhos e camundongos, porém a doença é reproduzida de forma mais consistente em coelhos. Portanto, sugere-se que coelhos possam ser utilizados como modelos para estudos in vivo com o ORFV.
Coelho, João Nuno Santos. "Molecular characterization and functional analysis of ORF P1192R from African swine fever virus." Doctoral thesis, Universidade de Lisboa. Faculdade de Medicina Veterinária, 2016. http://hdl.handle.net/10400.5/10907.
Full textAfrican swine fever virus (ASFV) is a nucleo-cytoplasmic large DNA arbovirus and the single member of the family Asfarviridae. It infects soft ticks of the genus Ornithodoros as well as all members of the family Suidae, representing a global threat for pig husbandry for which there is currently no effective vaccine or treatment. Since the ASFV viral cycle is mainly cytoplasmic, it has been found/predicted to code for many components of the replicative and transcriptional machineries. Of these, and based in sequence homologies, a putative type II DNA topoisomerase-coding ORF (P1192R) was identified in the ASFV genome. DNA topoisomerases are enzymes that modulate the topological state of DNA molecules. They are ubiquitous and essential, participating in processes such as DNA replication, recombination and repair and also in transcription. Since ASFV has a large linear genome, with 170 to 190 kbp depending on the isolate, containing terminal inverted repeats and covalently closed ends, a type II topoisomerase may be indispensable for viral replication and/or transcriptional events. The main objectives of this work were to deepen the study on ORF P1192R and determine if it indeed codes for a type II DNA topoisomerase and, if so, to characterize its activity. Bioinformatics and phylogenetic analyses showed that ORF P1192R is highly conserved among the fourteen ASFV isolates analyzed and, although its amino acid sequence clearly diverges from other type II topoisomerases, the structural organization is preserved and conserved motifs and domains essential for activity are present. Transient expression of GFP-pP1192R in COS-7 cells revealed an exclusively cytoplasmic distribution of the protein, which remained unaltered by treatment with leptomycin B. Using Vero cells or swine macrophages infected with ASFV isolate Ba71V or L60, respectively, expression of pP1192R was observed in the late phase of infection, co-localizing with the viral factories, where the bulk of viral replication and transcription occurs. Heterologous expression of pP1192R in Saccharomyces cerevisiae demonstrated that it functionally complements a top2 thermo-sensitive mutation and that it exhibits ATP-dependent decatenation activity. The purified recombinant pP1192R was found to efficiently decatenate kDNA and to processively relax supercoiled plasmid DNA, which are characteristics of a type II topoisomerase. The optimal requirements in terms of pH, temperature and salt, divalent ions and ATP concentrations for pP1192R activity in vitro were determined and its sensitivity to a panel of topoisomerase poisons and inhibitors was tested. Our results indicate that P1192R may be a target for studying, and possibly controlling, ASFV transcription and replication.
RESUMO - O vírus da peste suína africana (VPSA) é um arbovírus icosaédrico núcleo-citoplasmático de DNA de cadeia dupla, classificado no género Asfivirus da família Asfarviridae, da qual é o único membro conhecido. Este vírus infecta carraças do género Ornithodoros assim como todos os membros da família Suidae, constituindo uma ameaça global para a suinicultura para a qual não existe actualmente qualquer vacina ou tratamento. A prevenção da peste suína africana é feita através de medidas que visam reduzir o risco de introdução de animais ou produtos de origem animal infectados em regiões livres da doença, enquanto o controlo de um surto se baseia exclusivamente em medidas que incluem o abate sanitário de todos os animais susceptíveis na área do foco e a proibição de movimentos e comercialização de animais. Embora o VPSA tenha sido inicialmente descrito como um vírus com replicação exclusivamente citoplasmática, actualmente sabe-se que o núcleo da célula hospedeira é indispensável na fase inicial da infecção. Contudo, a grande maioria do ciclo infeccioso ocorre no citoplasma da célula infectada, não sendo por isso surpreendente que, das 150 a 167 grelhas de leitura aberta (ORF, do inglês “open reading frame”) identificadas no genoma do VPSA, algumas codifiquem para componentes das maquinarias de replicação e de transcrição. Dentre estas, prevê-se, com base em homologia de sequências aminoacídicas, que a ORF P1192R codifique para uma topoisomerase de DNA do tipo II. As topoisomerases de DNA estão presentes em todas as células e são responsáveis pela modulação do estado topológico do DNA, estado esse que se altera durante processos como a replicação, a recombinação e a reparação do DNA, assim como a transcrição, e dos quais resultam torções das moléculas de DNA que, não sendo resolvidas, podem comprometer a integridade genómica e consequentemente a viabilidade celular. Todas as topoisomerases exercem a sua actividade através da criação de quebras no DNA devido ao ataque nucleofílico de um resíduo de tirosina catalítico ao esqueleto fosfodiéster da molécula de DNA, gerandose assim uma ligação fosfotirosina covalente. As topoisomerases são classificadas em dois tipos, tendo por base a forma como quebram a molécula de DNA: as topoisomerases do tipo I, cuja actividade é independente de ATP e que geram quebras em cadeia única no DNA, facilitando assim o desenrolamento; e as topoisomerases do tipo II, que necessitam de ATP para gerar uma quebra nas duas cadeias do DNA, através da qual fazem passar uma dupla cadeia intacta. Considerando que o VPSA tem um genoma linear de grandes dimensões, com 170 a 190 quilopares de bases dependendo do isolado, e que contém repetições terminais invertidas fechadas covalentemente, uma topoisomerase do tipo II pode efectivamente ser essencial para eventos de replicação e/ou transcrição virais. Os objectivos centrais deste trabalho foram os seguintes: (i) realização de um estudo bioinformático e filogenético aprofundado da ORF P1192R do VPSA; (ii) estudo da proteína codificada por esta ORF (pP1192R), através da sua clonagem, expressão em sistema heterólogo, purificação da proteína recombinante e caracterização in vitro da sua actividade; (iii) determinação do efeito sobre a actividade da proteína recombinante dum painel de compostos químicos descritos como sendo inibidores de topoisomerases; (iv) identificação dos níveis de expressão e da localização intracelular da pP1192R em células infectadas pelo VPSA, a diferentes tempos de infecção; (v) avaliação do efeito de mutações dirigidas em resíduos ou motivos identificados como reguladores da actividade enzimática ou localização subcelular da pP1192R, tendo por base a informação gerada nos estudos bioinformáticos acima mencionados. A ORF P1192R do isolado L60 do VPSA foi amplificada por PCR e clonada e a sua sequência nucleotídica foi determinada e utilizada em análises bioinformáticas e filogenéticas. Verificou-se que esta ORF é altamente conservada entre os catorze isolados do VPSA cujo genoma se encontrava disponível nas bases de dados e, embora a sua sequência aminoacídica seja claramente divergente das de outras topoisomerases do tipo II incluídas neste estudo, quer sejam elas de origem procariota, eucariota ou viral, a organização estrutural da proteína está preservada e estão presentes motivos e domínios conservados que são essenciais para a actividade enzimática. O estudo da localização celular da pP1192R iniciou-se com a construção de plasmídeos quiméricos para a expressão da pP1192R em fusão com a proteína verde fluorescente (GFP) ou com uma variante vermelha (RFP). Transfectaram-se transientemente células de linha COS-7 com estas construções tendo-se observado que a proteína de fusão se distribuía exclusivamente pelo citoplasma. Esta distribuição não foi alterada após tratamento com leptomicina B que bloqueia uma das vias de exportação de proteínas do núcleo. Já a infecção das células a expressarem GFP-pP1192R com um isolado do VPSA adaptado a células Vero (Ba71V) induziu uma redistribuição da proteína de fusão, deixando de estar homogeneamente distribuída pelo citoplasma para estar principalmente concentrada nas fábricas virais a partir das 8 horas pós-infecção. Utilizando células de linha Vero infectadas com o isolado Ba71V, utilizado como modelo de infecção, ou macrófagos derivados de monócitos de sangue periférico de suíno (células alvo do vírus na infecção natural) infectados com o isolado virulento L60, e utilizando um soro anti-pP1192R produzido no decurso destes trabalhos, foi possível constatar que a pP1192R viral é produzida na fase intermédia/tardia da infecção (observável a partir das 6/8 horas pós-infecção) e que acumula nas fábricas virais ao longo da infecção. A expressão em sistema heterólogo da pP1192R iniciou-se num sistema procariota, baseado em Escherichia coli, mas embora tenha sido possível obter proteína recombinante em grandes quantidades, a sua purificação só foi conseguida recorrendo a agentes desnaturantes, impedindo a obtenção de proteína activa. Assim, avançou-se para um novo sistema de expressão baseado na levedura Pichia pastoris que apresenta diversas vantagens sobre o anterior, nomeadamente o facto de ser um sistema eucariota e por isso mais semelhante ao contexto em que a pP1192R é expressa em condições naturais. Contudo, neste sistema não foi possível obter proteína recombinante e o sistema foi abandonado. Tentou-se por fim a expressão heteróloga na levedura Saccharomyces cerevisiae. Neste organismo, a utilização das estirpes JCW26 e SD117 que contêm uma mutação termo-sensível no gene que codifica para a topoisomerase do tipo II endógena, permitiu demonstrar, quer in vivo através da complementação da mutação termo-sensível, quer in vitro recorrendo a ensaios funcionais de decatenação, que a pP1192R é efectivamente uma topoisomerase do tipo II funcional. Utilizando ainda S. cerevisiae como sistema de expressão, foi possível obter e purificar pP1192R recombinante para caracterização da sua actividade em ensaios funcionais in vitro. Observou-se que a pP1192R é capaz de relaxar DNA superenrolado, de decatenar DNA catenado e, quando em elevadas concentrações, de catenar DNA plasmídico, não tendo sido detectada actividade de superenrolamento de DNA relaxado. Determinaram-se também as condições óptimas de funcionamento em termos de temperatura, pH e concentrações de sal (NaCl ou KCl), ATP ou iões divalentes (Mg2+, Mn2+, Zn2+, Cu2+ e Ca2+), que foram posteriormente utilizadas para avaliar a sensibilidade da pP1192R recombinante a um painel de inibidores de topoisomerases, entre os quais se incluem drogas frequentemente utilizadas como agentes antimicrobianos ou antitumorais. Dos compostos testados, aqueles para os quais foram obtidos resultados mais promissores, i.e., os que revelaram níveis de inibição mais elevados, foram a coumermicina A1, a doxorubicina, a amsacrina e a genisteína. Pelo contrário, as quinolonas, normalmente utilizadas como antibióticos visando infecções provocadas por organismos procariotas, foram dos compostos com menor eficácia. Em suma, os resultados deste trabalho indicam que a ORF P1192R é um alvo promissor para o estudo e, eventualmente, o controlo dos processos replicativos e transcricionais do vírus da peste suína africana.
Housawi, Fadhel Mohammed Taher. "Studies on parapoxvirus antigens through the development of monoclonal antibodies to orf virus." Thesis, University of Edinburgh, 1998. http://hdl.handle.net/1842/30287.
Full textDurantel, David. "Etude des gènes pnk/pnl (ORF 86), lef-4 (ORF 90) et lef-5 (ORF 99) du virus de la polyédrose nucléaire de la tordeuse de la luzerne, "Autographa californica" (AcMNPV) : implication dans l'expression des gènes tardifs." Montpellier 2, 1997. http://www.theses.fr/1997MON20225.
Full textOnwuka, S. K. "Studies of orf virus replication and the cutaneous cellular response to infection in sheep." Thesis, University of Edinburgh, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.257195.
Full textTan, Joanne Li-Ching, and n/a. "Development of Orf virus as a vaccine vector : manipulation of structural proteins for surface display of immunogenic peptides." University of Otago. Department of Microbiology & Immunology, 2009. http://adt.otago.ac.nz./public/adt-NZDU20090427.144304.
Full textDeane, David Leslie. "The characterisation of a GM-CSF and IL-2 inhibitory protein encoded by Orf virus." Thesis, University of Edinburgh, 2001. http://hdl.handle.net/1842/23323.
Full textKomar, Monica. "Potentiating the Oncolytic Efficacy of Poxviruses." Thèse, Université d'Ottawa / University of Ottawa, 2012. http://hdl.handle.net/10393/23114.
Full textAnziliero, Deniz. "Efeitos do parapoxvirus ovis inativado sobre eventos da resposta imune inata em camundongos." Universidade Federal de Santa Maria, 2013. http://repositorio.ufsm.br/handle/1/4090.
Full textThe immunostimulatory properties of inactivated parapoxvirus ovis (iPPVO) have been investigated in different animal species and experimental settings. This study investigated the effects of administration of iPPVO on selected events of the innate response in mice. Neutrophil activation, phagocytic activity of macrophages, serum bactericidal activity, induction and antiviral activity of interferon type I (IFN - I) and expression of several classes of cytokines were assayed following intraperitoneal inoculation of Mus musculus with iPPVO (107 TCID50). Serum from iPPVO-treated animals showed IFN-I activity against murine encephalomyocarditis virus (EMCV) between 6 and 12 hours post infection (hpi), as shown by plaque reduction. A significant activation of neutrophils at 6hpi was observed by NBT reduction test in animals treated with the iPPVO. Peritoneal macrophages from mice treated with iPPVO demonstrated a significant increase (p<0.01) in phagocytic activity against Candida albicans both in vivo (between 12 and 96 hpi) and in vitro (24 and 72 hpi). iPPVO treated mice showed increased serum bactericidal activity against Escherichia coli (p<0.05) at two periods (24 and 72 hpi). A second study evaluated the expression of cytokines in response to inoculation of iPPVO. For this, spleens and serum samples were collected from mice treated with iPPVO at different intervals after inoculation and subjected to quantification of messenger RNA (mRNA) by real time PCR (qRT-PCR) and detection/quantification of serum cytokines by ELISA. Quantification of mRNA identified a significant and transient increase in the expression of various cytokines, with variable magnitude and kinetics. mRNA expression of proinflammatory cytokines (IL-1β, TNF-α and IL-8) peaked at 24 hpi (5.4 times increase) and 48 hpi (3 and 10 times, respectively). A 15-fold increase in expression of INF-γ and 6-fold for IL-12 was observed at 48 and 24 hpi, respectively. An increase in the expression of self-regulatory cytokines (Th2) cells, especially IL-10 and IL-4 was detected at later periods (72 and 96 hpi) with peaks of 4.7 and 4.9 fold, respectively. The determination of the concentration of serum cytokines by ELISA showed an increase in IL-1β, TNF-α, IL- 12, IFN-γ and IL-10 with kinetics similar to that observed by qPCR, especially for IL-1 and INF-γ. In summary, these results demonstrate that inoculation with iPPVO stimulates transiently a number events associated with cellular and humoral innate immune responses. If taken together, these effects would likely contribute for the enhanced resistance to certain pathogens observed in animals treated with iPPVO.
As propriedades imunoestimulatórias do Parapoxvirus ovis inativado (iPPVO) têm sido verificadas em diferentes espécies animais e condições experimentais. No presente trabalho foram investigados os efeitos da administração do iPPVO sobre eventos da resposta inata de camundongos. Ativação de neutrófilos, atividade fagocítica de macrófagos, atividade bactericida do soro, indução e atividade antiviral do interferon tipo I (INF-I) e expressão de várias classes de citocinas foram investigados em diferentes intervalos após inoculação de Mus musculus pela via intraperitonial com iPPVO (dose 107 TCID50). O soro de animais tratados com iPPVO apresentou atividade de INF-I frente ao vírus da encefalomiocardite murina (EMCV) entre 6 e 12 horas pós inoculação (hpi), como demonstrado pela redução significativa de formação de placas virais. Uma significativa ativação dos neutrófilos circulantes foi observada pela técnica de redução do NBT em animais tratados com o iPPVO às 6 hpi. Macrófagos peritoneais de camundongos tratados com iPPVO demonstraram um aumento significativo (p<0,01) na atividade fagocítica frente a Candida albicans tanto in vivo (entre 12 e 96 hpi) quanto in vitro (24 e 72hpi). Camundongos tratados com iPPVO apresentaram aumento na atividade bactericida do soro frente à Escherichia coli (p<0,05) em dois períodos avaliados (24 e 72 hpi). Um segundo estudo avaliou a expressão de citocinas em resposta à inoculação do iPPVO. Para isso, amostras de baço e soro foram coletados de camundongos tratados com iPPVO em diferentes intervalos após a inoculação e submetidas a quantificação de RNA mensageiro (RNAm) por PCR em tempo real (qRT-PCR) e detecção/quantificação de citocinas no soro por ELISA. A quantificação de RNAm permitiu detectar um aumento significativo e transitório da expressão de várias citocinas, com magnitude e cinética variáveis. A expressão de RNAm das citocinas pró-inflamatórias (IL-1β, TNF-α e IL-8) atingiu o pico às 24 hpi (aumento de 5,4 vezes), 48 hpi (3 e 10 vezes, respectivamente). Um aumento de 15 vezes na expressão gênica do INF-γ, e de 6 vezes para a IL-12 foi observado às 48 e 24 hpi, respectivamente. Um incremento na expressão das citocinas auto-regulatórias (Th2), principalmente IL-10 e IL-4, foi detectado em períodos mais tardios (72 e 96 hpi) com picos de 4,7 e 4,9 vezes, respectivamente. A determinação da concentração das citocinas séricas por ELISA revelou um aumento nos níveis de IL-1β, TNF- α, IL-12, INF-γ e IL-10, com uma cinética similar à observada pela técnica de qPCR, especialmente para IL-1β e INF-γ. Em resumo, esses resultados demonstram que o tratamento com iPPVO estimula de forma significativa e transitória uma série de eventos celulares e humorais ligados à resposta imune inata. Esses efeitos, se considerados em conjunto, provavelmente contribuem para o aumento da magnitude da resposta imunológica a certos patógenos observada em animais tratados com o iPPVO.
Rohde, Jörg [Verfasser], and Hans-Georg [Akademischer Betreuer] Rammensee. "Herstellung neuer Orf Virus- (Parapoxvirus) Rekombinanten und Analyse deren protektiven und immunrelevanten Eigenschaften / Jörg Rohde ; Betreuer: Hans-Georg Rammensee." Tübingen : Universitätsbibliothek Tübingen, 2017. http://d-nb.info/1199545694/34.
Full textBarros, Danielle Ribeiro de. "Clonagem e expressão in vitro da ORF AC5 do Tomato rugose mosaic virus (ToRMV) e produção de anti-soro." Universidade Federal de Viçosa, 2003. http://www.locus.ufv.br/handle/123456789/10116.
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O Tomato rugose mosaic virus (ToRMV) é um begomovírus descrito como parte de um complexo viral causando mosaico dourado e deformação (rugosidade) foliar em tomateiro na região do Triângulo Mineiro, Minas Gerais. Os dois componentes genômicos encontram-se completamente sequenciados, e a análise das seqüências de nucleotídeos do DNA-A indicou a presença de uma ORF adicional denominada AC5, presente apenas em algumas espécies de begomovírus. Esta ORF está quase totalmente inserida na sequência do gene cp, porém em orientação inversa e em outra fase de leitura. A ORF AC5 do ToRMV tem o potencial de codificar uma proteína com aproximadamente 27 kDa. A função do produto potencial da ORF AC5 foi estudada apenas para o Watermelon chlorotic stunt virus (WmCSV), não sendo encontrado nenhum indício da expressão da ORF. Entretanto, a seqüência de aminoácidos da proteína potencialmente codificada pela ORF AC5 do ToRMV possui apenas 41% de identidade com a AC5 do WmCSV. É possível que a ORF AC5 do ToRMV seja expressa, e que a proteína desempenhe alguma função no ciclo de infecção viral. Este trabalho teve por objetivo a clonagem e a expressão in vitro da ORF AC5 do ToRMV e a producão de anti-soro específico. Oligonucleotídeos específicos foram utilizados para a amplificação da região codificadora da ORF AC5 via PCR. O fragmento amplificado foi clonado no vetor de expressão pRSET-C. Plasmídeos recombinantes foram utilizados para a expressão da proteína AC5 em E. coli BL21::DE3. A proteína foi purificada a partir das células de E. coli e utilizada para a produção de anti-soro policlonal em coelhos. A especificidade do anti-soro policlonal obtido foi confirmada por Western blot. Esse anti-soro poderá ser utilizado em ensaios de imunolocalização da proteína AC5 em tecidos infectados pelo ToRMV.
Tomato rugose mosaic virus (ToRMV) is a typical bipartite begomovirus described as part of a viral complex inducing golden mosaic and leaf distortion (rugosity) in tomatoes at Triângulo Mineiro, Minas Gerais. Both genomic components have been completely sequenced, and the DNA-A sequence analysis indicated the presence of an additional open ready frame (ORF), named AC5, present in only a number of begomovirus species. This ORF is almost fully inserted into the cp gene, but in opposite orientation and in a different reading frame. The AC5 ORF of ToRMV has the potential to encode for a 27 kDa protein. The function of a putative AC5 protein has only been studied for Watermelon chlorotic stunt virus (WmCSV), but no evidence of its expression was found. However, the amino acid sequence of the putative AC5 protein from ToRMV is only 41% identical to the WmCSV AC5. It is thus conceivable that the ToRMV AC5 is expressed and plays a role in the viral infection cycle. The objective of this work was to clone and express the ToRMV AC5 ORF in vitro, and to raise an AC5-specific antiserum. Specific primers were used to direct the PCR-based amplification of the ToRMV AC5 ORF. The amplified fragment was cloned into the expression vector pRSET-C. Recombinant plasmids were used for protein expression in E. coli BL21::DE3. The AC5 protein was purified from E. coli cells by affinity chromatography and used for the immunization of rabbits. The specificity of the polyclonal antiserum obtained was assayed by Western blot. This antiserum can now be used in immunolocalization studies attempting to detect the AC5 protein in ToRMV-infected tissues.
Dissertação importada do Alexandria
LEH, VERONIQUE. "Etude des interactions entre proteines engageant les produits des orf iii et vi du virus de la mosaique du chou-fleur." Université Louis Pasteur (Strasbourg) (1971-2008), 1999. http://www.theses.fr/1999STR13180.
Full textScholz, Kai. "Immunmodulation durch Parapocken-Viren: Identifikation und Analyse funktionaler Viruskomponenten." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2003. http://nbn-resolving.de/urn:nbn:de:swb:14-1061969873968-07930.
Full textLi, Hua. "RNA SEQUENCE DETERMINANTS OF A COUPLED TERMINATION-REINITIATION STRATEGY FOR TRANSLATION OF DOWNSTREAM ORF IN HELMINTHOSPORIUM VICTORIAE VIRUS 190S AND OTHER VICTORIVIRUSES (FAMILY TOTIVIRIDAE)." UKnowledge, 2014. http://uknowledge.uky.edu/plantpath_etds/9.
Full textBhatta, Ankit. "Role of a Mitochondrial Micropeptide in Regulating Innate Immune Responses." eScholarship@UMMS, 2020. https://escholarship.umassmed.edu/gsbs_diss/1108.
Full textBentley, Emma. "The study of highly pathogenic emerging zoonotic virus envelope proteins through pseudotyped virus generation." Thesis, University of Westminster, 2017. https://westminsterresearch.westminster.ac.uk/item/q4yzx/the-study-of-highly-pathogenic-emerging-zoonotic-virus-envelope-proteins-through-pseudotyped-virus-generation.
Full textRintoul, Julia. "ORFV: A Novel Oncolytic and Immune Stimulating Parapoxvirus Therapeutic." Thèse, Université d'Ottawa / University of Ottawa, 2012. http://hdl.handle.net/10393/22925.
Full textYadav, Kush Kumar. "Genotype 1 hepatitis E virus (HEV) ORF4 protein enhances genotype 3 HEV replication." The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1574781581580768.
Full textBagdassarian, Eugénie. "Interaction du virus de l'hépatite E avec la réponse intérféron de l’hôte." Thesis, Université Paris-Saclay (ComUE), 2017. http://www.theses.fr/2017SACLS266/document.
Full textHepatitis E virus (HEV) is the causative agent of acute hepatitis in humans that can lead to fulminant hepatitis in 1-4% of cases, and in 20% of pregnant women in endemic regions. HEV is an enterically-transmitted virus responsible for large waterborne epidemics in developing countries and numerous cases of zoonotic hepatitis E in industrialized countries. The recent description of cases of chronic hepatitis E and severe neurological disorders highlight the importance to characterize the interactions between HEV and the host. The objective of this PhD project was to characterize the interactions between HEV and the host innate immune response and particularly with the type I interferon system (IFN-I).The first part of this project aimed to study the ability of the ORF1 non-structural polyprotein of HEV to modulate the IFN-I signalling pathways. HEV ORF1 contains several putative functional domains including a methyltransferase (Met) and a papain-like cysteine protease (PCP) whose functions on the IFN-I signalling remain poorly understood. The results obtained showed that the MetPCP domain of ORF1 inhibits IFN-β and IFN-stimulated response element (ISRE) promoter activation and the expression of some IFN-stimulated genes (ISGs). We then investigated the mechanism involved in this inhibition of ISRE promoter activation. We showed that the MetPCP domain inhibits STAT1 phosphorylation and nuclear translocation. In contrast, we found that the MetPCP domain does not inhibit STAT2 phosphorylation. However, the mode of action of MetPCP remains to be fully characterised. The second part of this project aimed to determine the ability of plasmacytoid dendritic cells (pDCs) to produce IFN-I in response to HEV infection. Indeed, pDCs are the main IFN-I source and play a crucial role in innate and adaptive responses. The results obtained suggest that pDCs produce IFN-I moderately when co-cultured with HEV-infected cells. Taken together, the results obtained during this PhD project suggest that HEV has evolved different mechanisms to modulate the IFN-I host response
Kannan, Harilakshmi. "Functional characterization of the interaction of hepatitis E virus ORF3 product with the cytoskeleton." College Park, Md.: University of Maryland, 2008. http://hdl.handle.net/1903/8969.
Full textThesis research directed by: Virginia-Maryland Regional College of Veterinary Medicine. Maryland Campus. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.
Barkley, Russell. "Investigation of an Oncolytic MeV Cell-Cell Fusion Phenomenon Induced by an siRNA." Thesis, Université d'Ottawa / University of Ottawa, 2020. http://hdl.handle.net/10393/41531.
Full textHeidari, Farshad. "In vitro modulation of host immune response by the Varicella zoster virus ORF1 gene product." Thesis, Kingston University, 2017. http://eprints.kingston.ac.uk/41971/.
Full textAnkavay, Maliki. "Étude des modifications post-traductionnelles de la protéine de capside ORF2 du virus de l'hépatite E (HEV)." Thesis, Lille 2, 2019. http://www.theses.fr/2019LIL2S013.
Full textHepatitis E virus (HEV) infection is a major public health problem that affects more than20 million people and kills approximately 70,000 worldwide each year. HEV is the leadingcause of acute viral hepatitis in the world. In France, the seroprevalance of HEV is 22.4% inthe general population. This virus is transmitted by fecal-oral route or by consumption ofundercooked contaminated meat. Very recently, we have described an efficient cell culturesystem to amplify HEV. This system represents a unique tool for characterizing the variousstages of the infectious HEV lifecycle that are very poorly understood to date. As part of mythesis, I focused on the ORF2 capsid protein which is the structural unit of viral particles andis therefore a central player in the HEV lifecycle. ORF2 is a 660 amino acids protein thatcontains a signal peptide and three potential N-glycosylation sites (N1, N2 and N3). My workaimed to identify the role of the ORF2 N-glycosylation in the HEV lifecycle. Our resultsrevealed that the ORF2 protein is N-glycosylated on the N1 and N3 sites, whereas the N2 siteis not N-glycosylated. Also, the N-glycosylation has no significant relevance neither for thestability, oligomerization, antibody recognition of the ORF2 protein nor for viral assemblyand viral infectivity. We also revealed that, the ORF2 protein is translocated into the nucleusof infected cells, independently of N-glycosylation. We identified for the first time afunctional nuclear localization signal (NLS) located at the N-terminus of the ORF2 proteinthat interacts probably with the importin alpha1. Surprisingly, this NLS regulates both nuclearimport and endoplasmic reticulum translocation of the ORF2 protein. Finally, wedemonstrated that, the ORF2 protein possesses three nuclear export signals (NES) located atits C-terminus. The ORF2 protein interacts with the exportin1 and is exported from the cellnuclei. This interaction drives the ORF2 protein from the nucleus to the cytoplasm of theinfected cells. Hence, this study led to new insights into the molecular mechanisms of theORF2 protein and could help to the design of novel antiviral strategies against HEV
Blignaut, Marguerite. "The molecular and biological characterisation of ORF5 of three South African variants of Grapevine Vitivirus A." Thesis, Stellenbosch : University of Stellenbosch, 2009. http://hdl.handle.net/10019.1/2421.
Full textGrapevine Vitivirus A (GVA), genus Vitivirus, family Flexiviridae is a well characterised single-stranded RNA virus that has been implicated in the grapevine diseases, Kober stem grooving and Shiraz disease. The virus infects both its host, Vitis vinifera and the experimental model plant, Nicotiana spp.. Biological studies performed on the virus in its herbaceous host, Nicotiana benthami- ana, revealed that many divergent variants of the virus exists in South Africa and can induce di erent symptoms in the model plant. Further molecular analysis divided the variants into three molecular groups based on molecular heterogeneity and nucleotide identity. The establishment of an infectious full-length cDNA clone of GVA contributed towards the elucidation of gene functions for 4 of the 5 open reading frames (ORF's), and indicated ORF5 as the pathogenicity determinant within the genome. Further studies also showed that ORF5 encodes for a nucleic acid binding protein that exhibits suppression activity of a plants' natural virus silencing mechanism. Many proteins that have previously been identi ed as the pathogenicity determinant within a viral genome have been found to encode for suppression activity. Although suppression activity has been elucidated within the ORF5 of the Italian cDNA clone of GVA, IS 151, no such study has yet been performed on the divergent South African variants of GVA. Three variants, GTR1-1, GTR1- 2 and GTG11-1, which represent each of the molecular groups (Group III, II and I), were selected for this study. The aim of this study was to visually elucidate suppression activity of RNA transgene silencing by the ORF5's of GTR1-1, GTR1-2 and GTG11-1 in a transient expression assays in transgenic N. benthamiana (line 16c). Pathogenicity studies for these variants were also performed. The ORF5 of the infectious full-length clone, GVA118, which can also serve as an expression vector, was deleted and provided with restriction enzyme sites into which the respective ORF5s and the marker genes, GFP and GUS could be cloned directionally. Infectivity, symptom development and systemic movement were compared between the di erent full length clones after co-in ltration in N. benthamiana. Preliminary results obtained in this study failed to visually indicate any suppression activity encoded by the ORF5 of GTR1-1, GTR1-2 and GTG11-1. The deletion of ORF5 within GVA118 was successful and rendered the infectious full length clone asymptomatic. Directional cloning of the ORF5 of GTR1-1 into the unique restriction enzymes provided previously, resulted in much milder symptoms than those observe for GTR1-2 and GTG11-1. No GFP and GUS accumulation could be detected. This study has established an infectious full-length cDNA clone, pBINSN-e35SGVA118 ORF5-1-1-pA, that can possibly induce much milder symptoms in the herbaceous host, N. benthamiana. This construct can be further characterised as a possible expression vector of foreign proteins in herbaceous hosts and grapevine.
Guo, Hailong. "Antigenic epitope composition and protectivity of avian hepatitis E virus (avian HEV) ORF2 protein and vertical transmission of avian HEV." [Ames, Iowa : Iowa State University], 2006.
Find full textMartinez-Soto, Eduan E. "Understanding the Role of Health Care Workers in a Trade-off Model between Contact and Transmission for Ebola Virus Disease." The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1467993935.
Full textBrusini, Jérémie. "Conséquences de l'incompatibilité végétative et de l'infection virale sur l'écologie et l'évolution de l'interaction Cryphonectria parasitica X Cryphonectria Hypovirus." Thesis, Bordeaux 1, 2009. http://www.theses.fr/2009BOR13826/document.
Full textVegetative incompatibility systems have been described in Fungi as controlling somatic fusion between conspecifics. For fungi, only fungi of the same vc type can fuse together. As other self recognition systems, this system involved high allelic diversity at specific genes. The issue of this work is to study the cause and effect relationship between the evolution of vegetative incompatibility systems and the selective pressure drove by cytoplasmic deleterious elements, transmitted during somatic fusion. Three problematics with three different approach were done : (1) a conceptual general framework on the evolution of self recognition systems (2) a theoretical work on the maintenance of vc type diversity by DCE and (3) an experimental work on the study of relationship between transmission and virulence in the C. parasitica/CHV host-parasite system. Ours results showed the key role of the permeability of the vegetative incompatibility barrier both for vc type diversity maintenance and on DCE prevalence. DCE would evolve toward avirulence in response to the transmission limitation by host incompatibility systems. Experimental work suggested a positive link between virulence and transmission in some population of CHV when host present a low vc type diversity, which could allow the evolution of the DCE toward an intermediate optimal virulence. This study would shed some light on mechanisms acting on the ecology and the evolution of fungi/DCE interaction which, according to our results, would be good study models for works on host-parasite systems
Mohamed, Mahgoub Mohamed Ahmed Mohamed. "Sporadic on/off switching of HTLV-1 Tax expression is crucial to maintain the whole population of virus-induced leukemic cells." Kyoto University, 2018. http://hdl.handle.net/2433/232139.
Full textAlsafi, Radi Taha M. "Generation of complex recombinant fowlpox virus 9 (FP9) encoding simian immunodeficiency virus (SIVmac239) sequences as a model HIV vaccine candidate." Thesis, University of Manchester, 2016. https://www.research.manchester.ac.uk/portal/en/theses/generation-of-complex-recombinant-fowlpox-virus-9-fp9-encoding-simian-immunodeficiency-virus-sivmac239-sequences-as-a-model-hiv-vaccine-candidate(1a015762-8dc2-4153-a586-d7fab88b9658).html.
Full textBardelli, Martino. "Strategies of viral multi-functional regulator proteins : adeno-associated virus Rep." Thesis, King's College London (University of London), 2016. https://kclpure.kcl.ac.uk/portal/en/theses/strategies-of-viral-multifunctional-regulator-proteins(6154fd50-eacd-4dbe-b456-bd08d2b13849).html.
Full textKey, Kijona Farthing. "Molecular characterization of the major envelope protein of porcine reproductive and respiratory syndrome virus (PRRSV) and evaluation of its use for a diagnostic assay, vaccine development, and the examination of quasispecies evolution." Diss., Virginia Tech, 2007. http://hdl.handle.net/10919/27282.
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Flight, William George. "The role of respiratory viruses in exacerbations of cystic fibrosis in adults." Thesis, University of Manchester, 2014. https://www.research.manchester.ac.uk/portal/en/theses/the-role-of-respiratory-viruses-in-exacerbations-of-cystic-fibrosis-in-adults(fb64b48e-1b68-48c1-9095-5fdf5e96f9b4).html.
Full textAlosaimi, Bandar. "Studies on potential co-operativity between different types of tumour virus." Thesis, University of Manchester, 2015. https://www.research.manchester.ac.uk/portal/en/theses/studies-on-potential-cooperativity-between-different-types-of-tumour-virus(5dea86d5-418d-46a0-943c-da7d53653423).html.
Full textMaurin, Cédric. "Synthèse d'inhibiteurs potentiels de l'intégrase du VIH-1." Lille 1, 2004. https://ori-nuxeo.univ-lille1.fr/nuxeo/site/esupversions/ce7057c5-a24c-410e-a5fa-31a72a812588.
Full textAdomavicius, Tomas. "Evaluation of the hepatitis B virus particle as a malaria vaccine carrier." Thesis, University of Manchester, 2015. https://www.research.manchester.ac.uk/portal/en/theses/evaluation-of-the-hepatitis-b-virus-particle-as-a-malaria-vaccine-carrier(bb9e157f-dc5c-4b81-9731-5ced1cbef8fe).html.
Full textSchultz, Patrick. "Etudes structurales du minichromosome du virus sv40 et de la chromatine cellulaire : approches en microscopie electronique." Université Louis Pasteur (Strasbourg) (1971-2008), 1987. http://www.theses.fr/1987STR13082.
Full textAuburn, Helen. "Evaluation of the role of Epstein-Barr virus and cellular gene expression in paediatric and adult transplant recipients." Thesis, King's College London (University of London), 2012. https://kclpure.kcl.ac.uk/portal/en/theses/evaluation-of-the-role-of-epsteinbarr-virus-and-cellular-gene-expression-in-paediatric-and-adult-transplant-recipients(d0044c58-3c67-48dd-9a78-f67821a2245a).html.
Full textIslam, Ayesha. "Interactions of human immunodeficiency virus type 1 with mucosal epithelial surfaces and Candida albicans." Thesis, King's College London (University of London), 2012. https://kclpure.kcl.ac.uk/portal/en/theses/interactions-of-human-immunodeficiency-virus-type-1-with-mucosal-epithelial-surfaces-and-candida-albicans(c8f83c82-f3af-469e-9790-8626f4b96d0f).html.
Full textGoka, Edward Anthony Chilongo. "Influenza A viruses dual and multiple infections with other respiratory viruses and risk of hospitalization and mortality." Thesis, University of Manchester, 2014. https://www.research.manchester.ac.uk/portal/en/theses/influenza-a-viruses-dual-and-multiple-infections-with-other-respiratory-viruses-and-risk-of-hospitalization-and-mortality(256eb122-a52a-4276-8dc1-28b5a2cc6662).html.
Full textEstmer, Nilsson Camilla. "Viral Control of SR Protein Activity." Doctoral thesis, Uppsala : Acta Universatis Upsaliensis : Univ.-bibl. [distributör], 2001. http://publications.uu.se/theses/91-554-5124-1/.
Full textWong, Kiing Aik. "Age related seroepidemiological survey of measles, mumps, rubella, varicella zoster, herpes simplex type 1 and 2 viruses." Thesis, University of Manchester, 2015. https://www.research.manchester.ac.uk/portal/en/theses/age-related-seroepidemiological-survey-of-measles-mumps-rubella-varicella-zoster-herpes-simplex-type-1-and-2-viruses(7188f535-e90e-4e18-a5cb-9b5ed1a75c68).html.
Full textNizard, Mevyn. "Optimisation d'un vaccin thérapeutique dans les tumeurs des voies aérodigestives supérieures associées aux papillomavirus : rôle de l'induction d'une immunité muqueuse et de la combinaison à la radiothérapie." Thesis, Sorbonne Paris Cité, 2015. http://www.theses.fr/2015PA05T027/document.
Full textCancer is the second mortality cause worldwide while mucosal cancers (lung, stomac, …) is the first mortality cause from. The majority of cancer vaccines against mucosal tumors have not given rise yet to significant clinical results. In this work we developed a strong immunotherapy based on the nontoxic subunit B from shiga toxin and showed for the first time that the localization of the immunization is crucial to induce potent and effective anti-tumoral responses. In a preclinical model a systemic immunization failed to induce a therapeutical protection against mucosal tumor challenge while intranasal immunization completely succeed. We identified a CD8 T lymphocyte population as a required cells in this protection and more precisely the T resident memory (Trm) cells. This Trm showed the classical CD103 phenotype as well as the CD49a which can play a specific role in the retention or the migration of this cells in the tumor tissue and might play a role in the survival. We also demonstrate that dendritic cells from the mucosal parenchyma was required to induce the CD49a expression on CD8 T cells while dendritic cells from the spleen was not. Our work shows that the Trm number as an impact in the anti-tumoral protection. We were able to reduce the Trm number in vivo using an anti-TGF-β antibody. This number diminution was correlated with a less efficient anti-tumoral protection. Patients with head and neck cancers are treated with radiotherapy. In this situation we showed that the combination of radiotherapy and our immunotherapy was associated with a better protection than radiotherapy alone or immunotherapy alone thanks to a vascular normalization. These results might rapidly lead to clinical trials and might open new ways to work with immunotherapies
Lingen-Stallard, Andrew. "Testing HIV positive in pregnancy : a study of women's experience and personal testimony following a positive human immunodeficiency virus (HIV) antibody test result during pregnancy." Thesis, University of Manchester, 2013. https://www.research.manchester.ac.uk/portal/en/theses/testing-hiv-positive-in-pregnancy-a-study-of-womens-experience-and-personal-testimony-following-a-positive-human-immunodeficiency-virus-hiv-antibody-test-result-during-pregnancy(b7f9438d-eda0-44fd-9191-15c87d3b4b85).html.
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