Characterization of a schistosome T cell-stimulating antigen (Sm10) associated with protective immunity in humans.
Parasite antigens that are strong T cell immunogens represent potential candidates for vaccines against pathogens susceptible to T cell-mediated immunity. We have previously shown that chromatographic fractions of schistosomula extracts contain components that are major T cell immunogen(s) in natural schistosome infections in humans and might contribute to the induction of human protective immunity against this parasite. In the present study, we report on the molecular cloning and on the biochemical characterization of the active components of these fractions. The screening of a schistosomula cDNA expression library with antibodies raised against the fractions allowed the cloning of a cDNA that hybridized to a 0.56-kb mRNA of schistosomula and adult worms. This cDNA contains an open reading frame of 267 base pairs (bp) which encodes a 10-kDa polypeptide. The analysis of the cDNA sequence revealed 70% homology with the sequences of previously reported proteins of unknown function. The native molecules in the active fractions were analyzed by mass spectrometry after additional purification by reverse phase high-performance liquid chromatography (HPLC). This procedure revealed two components in the fractions of molecular mass 10383 +/- 2 Da and 10401 +/- 9 Da. Both polypeptides stimulated immune T cells and yielded tryptic peptides whose sequences matched the sequence of the cloned molecule. These two polypeptides probably correspond to different post-translationally modified forms of the polypeptide encoded by the cloned cDNA.
Kohlstädt
S
S
INSERM U399/Laboratoire de Parasitologie-Mycologie, Faculté de Médecine, Marseille, France.
Couissinier-Paris
P
P
Bourgois
A
A
Bouchon
B
B
Piper
K
K
Kolbe
H
H
Dessein
A J
AJ
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth; D000947:Antigens, Helminth / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D002999:Clone Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D006801:Humans; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D018510:RNA, Helminth / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012550:Schistosoma mansoni / Q000235:genetics* / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000276:immunology / Q000517:prevention & control; D017386:Sequence Homology, Amino Acid; D013601:T-Lymphocytes / Q000276:immunology*; D014612:Vaccines / Q000302:isolation & purification
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D018510:RNA, Helminth; D012333:RNA, Messenger; D014612:Vaccines; C106491:antigen Sm10, Schistosoma