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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2010.tde-07072021-110246
Document
Author
Full name
Marcus Vinícius Gomes da Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2010
Supervisor
Committee
Arruda Neto, Eurico de (President)
Thiemann, Otavio Henrique
Tosi, Luiz Ricardo Orsini
Title in Portuguese
Estudos sobre a interação de Leishmania RNA virus 1-4 com Leishmania (Viannia) guyanensis
Keywords in Portuguese
Infectividade
Leishmania (V) guyanensis
Leishmania RNA virus 1-4
Patogenicidade
Abstract in Portuguese
Leishmaniavirus (LRV) é um vírus de RNA de fita dupla que infecta persistentemente algumas espécies de Leishmania, principalmente os da subespécie Viannia. Embora o impacto na saúde pública da leishmaniose nos trópicos seja significativa, muito pouco se sabe sobre os papéis possíveis, se houver, que LRV pode ter sobre a biologia da Leishmania, como por exemplo na infectividade e patogenicidade. Este estudo foi conduzido para avaliar a replicação de LRV1-4 durante o ciclo de crescimento de Leishmania (V.) guyanensis da cepa M4147, que é naturalmente infectada com LRV1-4 (M4147/LRV1-4). Cultivo de M4147/LRV1-4 em meio sólido permitiu o isolamento de um clone sem LRV1-4 (nomeado clone 40), que foi utilizado como controle negativo. Não houve diferença entre as curvas de crescimento de M4147/LRV1-4 e clone 40. A carga viral de LRV1-4 determinada por PCR em tempo real mostrou que não houve replicação viral durante a fase exponencial de crescimento do M4147/LRV1-4 no estado líquido, mas somente após a fase estacionária de Leishmania. Houve uma diminuição progressiva da carga de LRV 1-4 ao longo do tempo durante as passagens de M4147/LRV1-4 na cultura axênica. Comparação de infectividade de macrófagos peritoneais in vitro mostrou que o percentual de macrófagos infectados foi significativamente maior para M4147/LRV1-4 (51%) do que para o clone 40 (28%) em 96 horas após a infecção, mas não antes desse tempo. Não houve diferença significativa no número de amastigotas por macrófago infectado. O teste de patogenicidade em coxim plantar de hamster infectados com M4147/LRV1-4 e com o Clone 40 não apresentaram diferenças no tamanho da lesão. LRV 1-4 replica-se lentamente em Leishmania (V) guyanensis, sem efeito aparente sobre o ciclo de crescimento do parasita ou de sua patogenicidade na pata de hamster, porem sua presença aumentou a porcentagem de macrófagos peritoneais infectados in vitro, sem alteração da carga parasitária.
Title in English
Effects of infection by Leishmania RNA virus 1-4 on phenotypes of L. guyanensis
Keywords in English
Infectivity
Leishmania (V) guyanensis
Leishmania RNA virus 1-4
Pathogenicity
Abstract in English
Leishmaniavirus (LRV) is a double-stranded RNA virus that persistently infects some species of Leishmania, mainly those of the subspecies Viannia. While the public health impact of leishmaniasis in the tropics is significant, very little is known about possible roles, if any, that LRV may have on the biology of Leishmania, their infectivity and pathogenicity. These studies were conducted to measure replication of LRV1-4 during the growth cycle of Leishmania (V.) guyanensis of the strain M4147, which is naturally infected with LRV1-4 (M4147/LRV1-4). Growing of M4147/LRV1-4 on solid médium permitted isolation of one clone without LRV1-4 (named clone 40), which was used as a negative control. There was no difference between growth curves of M4147/LRV1-4 and clone 40. LRV1-4 load determined by real-time PCR did not increase significantly during the exponential phase of growth of M4147/LRV1-4 in liquid, but only after the stationary phase was reached. There was a Progressive decrease in the load of LRV 1-4 over time during passages of M4147/LRV1-4 in axenic culture. Comparison of peritoneal macrophage infectivity in vitro showed that the percentage of infected macrophages was significantly higher for M4147/LRV1-4 (51%) than for clone 40 (28%) at 96 hours post infection, but not before that time. No significant differences in numbers of amastigotes per infected macrophage were noted. The pathogenicity assay in hamster footpad infected with M4147/LRV1-4 or clone 40 showed no differences in size of lesion and parasite load. LRV1-4 seems to replicate slowly in Leishmania (V.) guyanensis, without apparent effect on the parasite growth cycle or its pathogenicity in the hamster footpad, but its presence increases the ffequency of mouse peritoneal macrophage in vitro infection by the parasite, without change in the parasite load.
 
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Publishing Date
2021-07-07
 
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