Molecular cloning and characterization of a novel Schistosoma japonicum "irradiated vaccine-specific" antigen, Sj14-3-3.
A Schistosoma japonicum cDNA coding for a full length S. japonicum 14-3-3 protein was obtained by antibody screening of an adult worm cDNA library using sera taken from mice vaccinated with UV-attenuated cercariae, which are capable of transferring high levels of passive immunity to this parasite. The deduced amino acid sequence consists of 254 amino acids and is highly homologous with 14-3-3 family of proteins from a variety of species (55-69% identity). The recombinant S. japonicun 14-3-3 protein (rSj14-3-3) was expressed and purified in pGEX/E. coli, and in Western blotting was strongly recognised by sera from mice, rats and bovines vaccinated with irradiated S. japonicum cercariae. Analysis of mRNA showed that Sj14-3-3 is expressed in sporocysts and adult worms, but not in cercariae, however mouse antisera against rSj14-3-3 recognised a 29 kDa native antigen in antigen preparations made from eggs, cercariae, schistosomula and adult worms of S. japonicum indicating that this antigen is present in all life-cycle stages. The presence of the native antigen in detergent extracts of intact schistosomula suggests that it is also present in the schistosomular tegument which is the most vulnerable target for immune attack. However, antisera against rSj14-3-3 did not recognise a similar band in S. mansoni or S. haematobium antigens, indicating that, like the UV-attenuated vaccines, this protein induced species-specific immune responses. Southern blot analysis suggested that there may exist more than one gene copy and/or polymorphism for Sj14-3-3. Immunoelectron microscopy confirmed that the native antigen is present throughout the body of adult worms including the tegument, but is less abundant in the muscles. The potential of rSj14-3-3 as a vaccine is now under further investigation.
Zhang
Y
Y
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, UK. y.zhang@lshtm.ac.uk
Taylor
M G
MG
McCrossan
M V
MV
Bickle
Q D
QD
D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000235:genetics* / Q000528:radiation effects; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D005260:Female; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology* / Q000528:radiation effects; D051379:Mice; D008808:Mice, Inbred CBA; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics* / Q000276:immunology* / Q000528:radiation effects; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000528:radiation effects; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology* / Q000648:ultrastructure; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014446:Tyrosine 3-Monooxygenase; D014612:Vaccines / Q000235:genetics / Q000528:radiation effects
D048948:14-3-3 Proteins; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D017931:DNA Primers; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D011506:Proteins; D011994:Recombinant Proteins; D014612:Vaccines; D014446:Tyrosine 3-Monooxygenase