Characterization of a clone from an adult worm cDNA library selected with anti-Schistosoma mansoni human antibodies dissociated from immune complexes: a preliminary report.
Considering the scarcity of defined antigens, actually useful and reliable for use in the field studies, we propose an alternative method for selection of cDNA clones with potential use in the diagnosis of schistosomiasis. Human antibodies specific to a protein fraction of 31/32 kDa (Sm31/32), dissociated from immune complexes, are used for screening of clones from an adult worm cDNA library. Partial sequencing of five clones, selected through this strategy, showed to be related to Schistosoma mansoni: two were identified as homologous to heat shock protein 70, one to glutathione S-transferase, one to homeodomain protein, and one to a previously described EST (expressed sequence tag) of S. mansoni. This last clone was the most consistently reactive during the screening process with the anti-Sm31/32 antibodies dissociated from the immune complexes. The complete sequence of this clone was obtained and the translation data yielded only one ORF (open reading frame) that code for a protein with 57 amino acids. Based on this amino acid sequence two peptides were chemically synthesized and evaluated separately against a pool of serum samples from schistosomiasis patients and non-schistosomiasis individuals. Both peptides showed strong reactivity only against the positive pool, suggesting that these peptides may be useful as antigens for the diagnosis of schistosomiasis mansoni.
Valli
Luiz Carlos Pedrosa
LC
Faculdade de Ciências Farmacêuticas/USP, São Paulo, SP, Brasil. valli@univix.br
Kanamura
Herminia Yohko
HY
Cotrim
Paulo Cesar
PC
Correa Oliveira
Guilherme
G
Oliveira
Edward José de
EJ
D000818:Animals; D000909:Antibodies, Helminth / Q000235:genetics / Q000276:immunology; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics* / Q000276:immunology; D020224:Expressed Sequence Tags; D015723:Gene Library; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology; D006801:Humans; D016366:Open Reading Frames; D019151:Peptide Library; D012550:Schistosoma mansoni / Q000235:genetics* / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000175:diagnosis*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D019151:Peptide Library