RESUMO
In this study, the authors have generated a tapeworm Taenia solium genomic DNA expression library where foreign peptides/proteins were fused to N-termini of M13 cpVIII and expressed at a high copy number on the phage surface, and they showed that this library may be used in bioselection against antipathogen immune sera, allowing the identification of disease-related antigens recognizing antibodies present in clinical samples. They isolated 2 phage clones expressing T. solium-derived antigens specifically reacting with antibodies present in plasma and cerebrospinal fluid samples of neuroimaging-confirmed neurocysticercosis patients. The described antigen discovery strategy may be used for the direct identification of antigens useful for host-pathogen interaction studies as well as for the development of molecular vaccines and diagnostics.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Anticorpos Anti-Helmínticos/líquido cefalorraquidiano , Anticorpos Anti-Helmínticos/imunologia , Neurocisticercose/imunologia , Taenia solium/imunologia , Animais , Anticorpos Anti-Helmínticos/genética , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Bacteriófago M13/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Biblioteca Genômica , Interações Hospedeiro-Parasita , Humanos , Imunoglobulina G/líquido cefalorraquidiano , Imunoglobulina G/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Neurocisticercose/diagnóstico , Biblioteca de Peptídeos , Taenia solium/genéticaRESUMO
A novel cDNA cloning strategy consisting in elimination of non-coding DNA sequences from 3' regions of cDNAs was applied to construct the Taenia crassiceps phage displayed cDNA expression library. After biopanning using immune sera, three phage clones expressing T. crassiceps-derived antigens specifically recognizing antibodies present in cerebrospinal fluid and plasma samples from neuroimaging-confirmed neurocysticercosis patients were selected. This novel cloning strategy may be applied to other pathogens allowing rapid identification of peptides/proteins for immunodiagnostic tests.