Molecular characterization of a 23-kilodalton major antigen secreted by Toxoplasma gondii.
The strategy chosen for cloning potential vaccine antigens of Toxoplasma gondii was based on the hypothesis that the definitive protection observed in natural infection is due to the presence of encysted bradyzoite forms in host tissues throughout life. The antigens released by the bradyzoites would maintain an immune response against the invading tachyzoites. This led us to identify in tachyzoite in vitro translation products a polypeptide of 24 kDa that is an excreted-secreted antigen (ESA) and is cross-reactive with bradyzoites. In addition, the detection of anti-P24 IgG antibodies is correlated with the chronic infection in man. The gene encoding P24 has been isolated, sequenced, and expressed in Escherichia coli and eukaryotic cells. The recombinant proteins were immunogenic in mice, producing anti-native P23 antibodies. Immunocytochemical analysis located the native antigen in the dense granules of both tachyzoite and bradyzoite forms and showed that it is secreted within host-cell-modified phagosome. Moreover 45Ca2+ labeling as well as regional homologies indicate that this protein has Ca2+-binding properties, suggesting its physiological importance in host-cell invasion. P23 is of diagnostic interest as a marker of chronic toxoplasmosis and is proposed as a vaccine component.
Cesbron-Delauw
M F
MF
Unité Mixte Institut National de la Sante et de la Recherche Medicale U167, Institut Pasteur, Lille, France.
Guy
B
B
Torpier
G
G
Pierce
R J
RJ
Lenzen
G
G
Cesbron
J Y
JY
Charif
H
H
Lepage
P
P
Darcy
F
F
Lecocq
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015723:Gene Library; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D014176:Protein Biosynthesis; D014122:Toxoplasma / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D014123:Toxoplasmosis / Q000276:immunology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; C412913:GRA1 antigen; D015345:Oligonucleotide Probes; D004247:DNA