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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2008.tde-03062008-190643
Document
Author
Full name
Misael Leonardo Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Harsi, Charlotte Marianna (President)
Pinho, João Renato Rebello
Rizzo, Luiz Vicente
Title in Portuguese
Interação do adenovírus humano, sorotipo 41, com células origem hematopoiética: análise da permissividade celular e da expressão gênica viral.
Keywords in Portuguese
Adenovírus
Cinética de infecção
Linfócitos intraepiteliais
PBMC
Permissividade celular
Abstract in Portuguese
Para verificar a permissividade de células de origem hematopoiéticas à infecção por HAdV-41, foram infectados PBMC e IEL de voluntários. Os ensaios foram comparados com células HEK-293 infectadas. Foram analisadas as expressões dos genes virais E1A, E1B (55K), E3 (14K), VARNA, hexon e fibra curta (FC) e do gene celular GAPDH. O mRNA foi detectado por RT-PCR em tempo real e a produção de proteínas foi visualizada por IFI. Em HEK-293, a transcrição dos genes E1A, E1B e E3 iniciou-se às 11h p.i, hexon,às 13h pi.,VARNA e FC às 14h p.i. Em PBMC a transcrição de E1A, E1B e VARNA iniciou-se 17h p.i e a expressão dos genes hexon e fibra curta foi detectada 18h p.i e 20 h p.i. respectivamente. O nível de expressão dos genes virais em HEK-293 foi quase 200 vezes maior em relação à PBMC. Os IELs também mostraram-se permissivos à infecção pelo HAdV-41 como mostrado pela expressão dos genes virais. Essa é a primeira evidência de que este vírus possa infectar tais células. Os resultados obtidos ajudam a elucidar os mecanismos de interação do vírus com a célula-hospedeira.
Title in English
Human adenovírus serotype 41 interaction with hematopoietic cells: cellular permissiviness and viral gene expression analysis.
Keywords in English
Adenoviruses
Cellular permissiveness
Infection kinetics
Intraepithelial lymphocytes
PBMC
Abstract in English
In order to verify the permissiveness of hematopoietic cells to HAdV-41 infection, PBMC and IEL from volunteers were infected. The infection assays were compared with infected HEK -293 cells. We analysed the E1A, E1B (55K), E3 (14K), VARNA, hexon and short fiber (SF) viral gene expression and GAPDH cellular gene expression. The mRNA were detected by real time PCR and the viral protein synthesis were detected by IIF. In HEK-293 cells E1A, E1B and E3 gene expression were detected 11h p.i Hexon gene expression was detected at 13h p.i, while VARNA and SF were detected 14h p.i In PBMC, E1A, E1B and VARNA gene expression were detected 17h p.i and the hexon and SF were detected 18h p.i and 20h p.i, respectively. The viral gene expression level in infected HEK-293 cells was 200 fold higher than infected PBMC. The IEL also were permissive to HAdV-41 infection showed by viral gene expressions. This is the first evidence that HAdV-41 is able to infect these cells types. These results helps to understand the virus-cell interaction mechanisms.
 
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Publishing Date
2008-06-11
 
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