Differential localisation of an Eimeria tenella aspartyl proteinase during the infection process.
Aspartyl proteinases are essential for the survival of many pathogens. A single copy gene in species of Eimeria encodes an aspartyl proteinase, which we propose should be called eimepsin to conform to the commonly used names of this family of proteinases. An epitope map, constructed using BIAcore technology, confirmed the specificity of 14 mAbs for eimepsin and defined four antigenic domains, which were conserved between native and recombinant forms of eimepsin. In resting sporozoites, mAb defining antigenic domains I and II stained the refractile body organelles, whereas those defining antigenic domains III and IV stained cytoplasmic granules. During host cell invasion, the staining patterns of mAb defining antigenic domains I, III and IV changed dramatically with the apical tips of invading sporozoites becoming strongly stained. In contrast, mAb defining antigenic domain II continued to stain only the refractile bodies. During early schizogony, mAb to all four domains stained the single fused refractile body, but when schizonts matured, mAb to antigenic domains I, III and IV stained the apical tip of merozoites whereas those to antigenic domain II continued to follow the developmental redistribution of the refractile body. Irrespective of localisation, mAb to three antigenic domains recognised a polypeptide of 49 kDa, which from N-terminal sequencing corresponds to a mature form of eimepsin. Staining with fluorescent pepstatin localised a mature, active form of eimepsin to the refractile bodies of the sporozoite, schizont and first generation merozoite. It remains to be determined whether eimepsin has a catalytic function within the refractile body or whether the activated enzyme is stored in the refractile body so that it can be rapidly redistributed to the apical tip during parasite invasion.
Jean
L
L
Unité de Virologie et d'Immunologie Moléculaires, INRA, Domaine de Vilvert, 78352, Cedex, Jouy-en-Josas, France.
Grosclaude
J
J
Labbé
M
M
Tomley
F
F
Péry
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D016282:Aspartic Acid Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003048:Coccidiosis / Q000469:parasitology*; D016786:Eimeria tenella / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity; D018604:Epitope Mapping; D019084:Fluorescent Antibody Technique, Indirect; D015388:Organelles / Q000201:enzymology; D010436:Pepstatins / Q000378:metabolism; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010436:Pepstatins; D011994:Recombinant Proteins; D016282:Aspartic Acid Endopeptidases