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Doctoral Thesis
DOI
10.11606/T.87.2016.tde-25082016-100745
Document
Author
Full name
Aline Rodrigues Florencio Teixeira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Nascimento, Ana Lucia Tabet Oller do (President)
Bertoncini, Michelle Darrieux Sampaio
Piazza, Roxane Maria Fontes
Ruiz, Rita de Cássia
Silva, Rosa Maria
Title in Portuguese
Avaliação e caracterização de candidatos vacinais voltados para o controle da leptospirose.
Keywords in Portuguese
Leptospira
Leptospirose
Patogenicidade
Proteínas recombinantes
Vacina
Abstract in Portuguese
A leptospirose é uma doença sistêmica, causada por bactérias patogênicas do gênero Leptospira. O desenvolvimento de novas estratégias para prevenir a doença é necessário. Vacinas surgem como fortes candidatas para contornar o problema. As pesquisas atuais têm interesse em identificar antígenos conservados que estão envolvidos nas interações patógeno-hospedeiro.O presente projeto selecionou três proteínas hipotéticas de L. interrogans para serem caracterizadas quanto ao seu papel na patogênese e avaliadas quanto ao seu potencial protetor. Os genes foram amplificados por PCR e clonados no vetor de expressão PAE. As proteínas recombinantes foram purificadas por cromatografia de afinidade e foram reconhecidas por soro de indivíduos infectados. As proteínas LIC13479 e LIC10050 foram capazes de se ligar a laminina, plasminogênio e fibronectina plasmática. Em relação à LIC10537, dois fragmentos recombinantes foram gerados. Apenas o fragmento 2 foi capaz de interagir com PLG. As proteínas que interagiram com o PLG foram capazes de gerar plasmina As proteínas foram capazes de estimular uma resposta imune e LIC13479 e LIC10050 exerceram proteção parcial no modelo de leptospirose em hamsters.
Title in English
Evaluation and characterization of vaccine candidates against leptospirosis.
Keywords in English
Leptospira
Leptospirosis
Pathogenesis
Recombinant proteins
Vaccine
Abstract in English
Leptospirosis is a systemic disease caused by pathogenic bacteria of genus Leptospira. The development of new strategies to prevent the disease is needed. Vaccines emerge as strong candidates to fight the problem.Currently research has focused to identify conserved antigens This project selected three hypothetical proteins of L. interrogans. Thesecoding sequences were characterized for their possible role in pathogenesis and their potential to protect animals against challenge with virulent leptospires. Genes were amplified by PCR and cloned into the expression vector pAE. The recombinant proteins were purified by metal affinity chromatography and were recognized by confirmed human leptospirosis serum samples.LIC13479 and LIC10050 proteins were able to bind with laminin, plasminogen and plasma fibronectin. The coding sequence LIC10537 was cloned in two fragments. Fragment 2was able to interact with plasminogen. All proteins were able to generate active plasmin. The recombinant proteins were able of inducing an immune response. Evaluation of immunoprotection in leptospirosis hamster model followed by challenge with virulent bacteria showed that the recombinant proteins conferred partial protection.
 
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Publishing Date
2016-08-25
 
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