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Background  Avian coccidiosis is a widespread, economically significant disease of poultry, caused by several  Eimeria  species. These parasites have complex and diverse life-cycles that require invasion of their host cells. This is mediated by various proteins secreted from apical secretory organelles. Apical membrane antigen 1 (AMA1), which is released from micronemes and is conserved across all apicomplexans, plays a central role in the host cell invasion. In a previous study, some putative  Et AMA1-interacting proteins of  E. tenella  were screened. In this study, we characterized one putative  Et AMA1-interacting protein,  E. tenella Eimeria  -specific protein ( Et Esp).    Methods  Bimolecular fluorescence complementation (BiFC) and glutathione S-transferase (GST) fusion protein pull-down (GST pull-down) were used to confirm the interaction between  Et AMA1 and  Et Esp  in vivo  and  in vitro.  The expression of  Et Esp was analyzed in different developmental stages of  E. tenella  with quantitative PCR and western blotting. The secretion of  Et Esp protein was tested with staurosporine when sporozoites were incubated in complete medium at 41 °C. The localization of  Et Esp was analyzed with an immunofluorescence assay (IFA). An  in vitro  invasion inhibition assay was conducted to assess the ability of antibodies against  Et Esp to inhibit cell invasion by  E. tenella  sporozoites.    Results  The interaction between  Et AMA1 and  Et Esp was confirmed with BiFC and by GST pull-down. Our results show that  Et Esp is differentially expressed during distinct phases of the parasite life-cycle. IFA showed that the  Et Esp protein is mainly distributed on the parasite surface, and that the expression of this protein increases during the development of the parasite in the host cells. Using staurosporine, we showed that  Et Esp is a secreted protein, but not from micronemes. In inhibition tests, a polyclonal anti-r Et Esp antibody attenuated the capacity of  E. tenella  to invade host cells.    Conclusion  In this study, we show that  Et Esp interacts with  Et AMA1 and that the protein is secreted protein, but not from micronemes. The protein participates in sporozoite invasion of host cells and is maybe involved in the growth of the parasite. These data have implications for the use of  Et AMA1 or  Et AMA1-interacting proteins as targets in intervention strategies against avian coccidiosis.

Eimeria tenella Eimeria-specific protein that interacts with apical membrane antigen 1 (EtAMA1) is involved in host cell invasion