Repertoire of Theileria equi immunodominant antigens bound by equine antibody

- Characterization of immunodominant antigens of T. equi.
- Immunoaffinity chromatography of T. equi merozoite proteins.
- Identification of T. equi merozoite proteins by mass spectrometry.
- Confirmation of antibody specificity using recombinant T. equi proteins.

Theileriosis in horses and cattle is caused by tick-borne Apicomplexa parasites and results in death or life-long infection in their respective hosts. Transmission risk associated with persistent infection severely limits movement of horses and cattle resulting in economic losses. The recent reemergence of Theileria equi infection in U.S. horses demonstrates the continual threat Apicomplexa parasites represent to global animal health. A paucity of data concerning equine immune responses to T. equi, including antigens recognized by antibodies in clinically asymptomatic, persistently infected horses, precludes vaccine development. Therefore, this investigation was initiated to characterize antigens recognized by the equine antibody response to T. equi. This goal was accomplished by defining T. equi merozoite antigens that are recognized by antibodies in horses infected with distinct T. equi isolates. Previously it was shown that equine post-infection serum consistently recognized at least five T. equi merozoite antigens, but their precise identity remained unknown. To determine specificity of antibody target identification, T. equi merozoite antigens were first isolated using equine post-infection serum in affinity chromatography. Proteins recognized by the equine antibodies were then isolated from two-dimensional electrophoresis gels, and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) using the recently available T. equi genome database. Five T. equi antigens were identified and include Equi Merozoite Antigen-2 (EMA-2), EMA-3 and EMA-6, a previously uncharacterized protein annotated as “signal peptide containing protein”, and 40S ribosomal protein S12.

Recognition and identification of T. equi merozoite immunodominant antigens by antibodies isolated from persistently infected horses.224