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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-23092008-153317
Document
Author
Full name
Letusa Albrecht
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Wunderlich, Gerhard (President)
Costa, Fábio Trindade Maranhão
Ferreira, Marcelo Urbano
Ho, Paulo Lee
Shaw, Jeffrey Jon
Title in Portuguese
Análise do repertório de genes variantes de Plasmodium falciparum da amazônia e identificação de genes variantes relacionados ao fenótipo de citoaderência a ICAM1 de isolados de Rondônia.
Keywords in Portuguese
Plasmodium falciparum
Aderência celular
Antígenos
Biologia molecular
Malária
Polimorfismo
Abstract in Portuguese
Nesta tese estudamos os repertórios das famílias multigênicas var (domínio DBLa), rif e stevor, importantes fatores de virulência, de Plasmodium falciparum em isolados brasileiros. Mostramos que estas famílias multigênicas podem ser conservadas entre diferentes isolados no Brasil, sugerindo que o repertório de genes variantes pode ser limitado em algumas localidades. Identificamos menos polimorfismos nos genes stevor que os genes var ou rif. Constatamos que genes var podem ser conservados ao longo do tempo em isolados brasileiros. Identificamos também os domínios DBLa e DBLb de genes var associados a citoaderência a CHOICAM1 em isolados de Rondônia. Mediante ensaios de ELISA, mostramos que domínios DBLbICAM foram reconhecidos de modo heterogêneo. Por citometria de fluxo, demonstramos que o parasita aderente em ICAM1 foi reconhecido majoritariamente pelo plasma de pacientes com infecção sintomática e menos de infecção assintomática. Além disso, identificamos por PCR em tempo real os genes var transcritos no clone 3D7 aderente em diferentes linhagens de células CHO.
Title in English
Repertoire analysis of variant genes from Plasmodium falciparum amazônia isolates and identification of variant genes related with cytoaderence on ICAM1 from Rondonia isolates.
Keywords in English
Plasmodium falciparum
Antigens
Cell adherence
Malaria
Molecular biology
Polimorfism
Abstract in English
Herein, we studied the repertoire of multigenic families of var (DBLa domain), rif and stevor from brazilian isolates of Plasmodium falciparum. We showed that these multigenic families could be conserved among different isolates from Brazil, suggesting that the repertoire of variant genes can be limited in certain areas. We identified less polymorphism in stevor genes than in var or rif genes. We also demonstrated that var genes can be conserved over time in Brazilian isolates and identified the DBLa and DBLb domains from var genes associated with cytoadherence to CHOICAM1 in Rondonian isolates. ELISA assays showed that these DBLbICAM domains were recognized in a heterogeneous fashion. By flow cytometry, we demonstrated that ICAM1-adherent parasites were stronger recognized by plasma from symptomatic infections than from non-symptomatic infections. Finally, we profiled the var gene transcripts in 3D7 parasites adherent to different CHO cells by real time PCR.
 
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Publishing Date
2008-09-24
 
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