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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2013.tde-07062013-152408
Document
Author
Full name
Henrique Krambeck Rofatto
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Leite, Luciana Cezar de Cerqueira (President)
Carboni, Beatriz Simonsen Stolf
Piazza, Roxane Maria Fontes
Winter, Carlos Eduardo
Wunderlich, Gerhard
Title in Portuguese
Caracterização molecular das nucleotídeo pirofosfatases/ fosfodiesterases de Schistosoma mansoni e investigação como antígenos vacinais.
Keywords in Portuguese
Schistosoma mansoni
Expressão gênica
Nucleotídeos
Tegumento animal
Vacinas
Abstract in Portuguese
Neste trabalho caracterizou-se a família das nucleotídeo pirofosfatases/ fosfodiesterases do parasita S. mansoni visando avaliá-los como antígenos vacinais. O parasita possui quatro proteínas desta família (SmNPP-5a, SmNPP-5b, SmNPP-5c e SmNPP-6), sendo que duas delas possuem maior expressão gênica nos estágios que infectam o homem. Dos anticorpos produzidos, apenas o anti-SmNPP-5a apresentou especificidade para a proteína nativa e utilizando-os demonstrou-se que a SmNPP-5a é uma glicoproteína associada às membranas do tegumento dos vermes adultos. Também se verificou que esses anticorpos inibiram parcialmente a atividade da enzima em parasitas vivos. Assim avaliamos a SmNPP-5a recombinante como antígeno vacinal juntamente com uma apirase (SmATPDase) e com a fosfatase alcalina (SmAP), outras duas nucleotidases presentes no tegumento de parasitas adultos. A SmNPP-5 e a SmNTDPase apresentaram menor imunogenicidade que a SmAP. Porém só verificamos a redução da carga parasitária em animais imunizados com a SmAP e tratados com doses subcurativas de praziquantel.
Title in English
Molecular characterization of nucleotide pyrophosphatases/ phosphodiesterases of Schistosoma mansoni and their investigation as vaccine antigens.
Keywords in English
Schistosoma mansoni
Gene expression
Integument animal
Nucleotides
Vaccines
Abstract in English
In this work the family of nucleotide pyrophosphatases/phosphodiesterases (NPP) from S. mansoni was characterized in order to evaluate them as vaccine antigens. The parasite has four proteins of this family (SmNPP-5a, SmNPP-5b, SmNPP-5c and SmNPP-6), whereas two of them present higher gene expression in stages that infect humans. Only anti-SmNPP-5a anitbodies showed specificity for the native protein. We use them to characterize SmNPP-5a as a glycoprotein associated with tegument membranes from adult worms. It was also found that these antibodies were able to partially inhibit the activity of the enzyme in live parasites. Thus we evaluated SmNPP-5a as recombinant vaccine antigen together with an apyrase (SmATPDase) and alkaline phosphatase (SmAP), two other nucleotidases involved in nucleotide metabolism and present in the tegument of adult parasites. SmNTDPase and SmNPP-5 were less immunogenic than SmAP. However, we only verified the reduction of parasite burden in mice immunized with SmAP, when the animals also received a subcurative treatment with praziquantel.
 
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Publishing Date
2013-06-13
 
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