Abstract
Clinical chicken coccidiosis is mostly caused by simultaneous infection of several Eimeria species, and host immunity against Eimeria is species-specific. It is urgent to identify common immunodominant antigen of Eimeria for developing multivalent anticoccidial vaccines. In this study, sporozoite proteins of Eimeria tenella, Eimeria acervulina and Eimeria maxima were analyzed by two-dimensional electrophoresis (2DE). Western bot analysis was performed on the yielded 2DE gel using antisera of E. tenella E. acervulina and E. maxima respectively. Next, the detected immunodominant spots were identified by comparing the data from MALDI-TOF-MS/MS with available databases. Finally, Eimeria common antigens were identified by comparing amino acid sequence between the three Eimeria species. The results showed that analysis by 2DE of sporozoite proteins detected 629, 626 and 632 protein spots from E. tenella, E. acervulina and E. maxima respectively. Western bot analysis revealed 50 (E. tenella), 64 (E. acervulina) and 57 (E. maxima) immunodominant spots from the sporozoite 2DE gels of the three Eimeria species. The immunodominant spots were identified as 33, 27 and 25 immunodominant antigens of E. tenella, E. acervulina and E. maxima respectively. Fifty-four immunodominant proteins were identified as 18 ortholog proteins among the three Eimeria species. Finally, 5 of the 18 ortholog proteins were identified as common immunodominant antigens including elongation factor 2 (EF-2), 14-3-3 protein, ubiquitin-conjugating enzyme domain-containing protein (UCE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In conclusion, our results not only provide Eimeria sporozoite immunodominant antigen map and additional immunodominant antigens, but also common immunodominant antigens for developing multivalent anticoccidial vaccines.
Highlights
Avian coccidiosis, a major parasitic disease of chickens worldwide, was caused by intestinal infection of Eimeria spp. [1]
The results showed that analysis by two-dimensional electrophoresis (2DE) of sporozoite proteins detected 629, 626 and 632 protein spots from E. tenella, E. acervulina and E. maxima respectively
Sporozoite 2DE gels of E. tenella, E. acervulina and E. maxima were analyzed by western blot using the corresponding antisera of these Eimeria species separately
Summary
A major parasitic disease of chickens worldwide, was caused by intestinal infection of Eimeria spp. [1]. A major parasitic disease of chickens worldwide, was caused by intestinal infection of Eimeria spp. Present control strategy against this disease relies on anticoccidial drugs and live vaccines containing virulent or attenuated strains of Eimeria [5]. Clinical coccidiosis is mainly caused by co-infection with multiple species of Eimeria [10, 11], a practical novel anticoccidial vaccine should contain the common antigens among Eimeria or antigens from multiple Eimeria species. Exploring immunodominant antigens, especially common antigens of Eimeria, is essential for developing novel vaccine against the simultaneous infection clinically
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