Secretion of a new spherical body protein of Babesia bovis into the cytoplasm of infected erythrocytes.

A cDNA encoding a new Babesia bovis spherical body protein 4 (BbSBP-4) was reported to have no significant homology to other apicomplexan proteins or previously reported B. bovis spherical body proteins. In the present study, we further examined the molecular characteristics of BbSBP-4 including the expression and cellular localization of the BbSBP-4. An anti-rBbSBP-4 mouse serum specifically reacted to a 41-kDa native protein B. bovis in Western blot analysis. The immunoelectron microscopic examination confirmed the localization of BbSBP-4 in spherical bodies, but not in the nucleus, rhoptries, and micronemes. Interestingly, the protein was found to be localized not only in the spherical body of B. bovis but also in the cytoplasm of infected erythrocytes (iRBC) at the later stage of parasite development. The confocal laser microscopic examination and Western blot analysis demonstrated the increased accumulation of BbSBP-4 in the cytoplasm of iRBC and in the supernatant of cultivated B. bovis during the late developmental stage of the parasite. These results suggest that BbSBP-4 was secreted from spherical body into cytoplasm of iRBC during the late developmental stage of the parasite before the rupture of infected RBC. Taken together, BbSBP-4 might play an important role as a secreted protein in the intracellular development and/or survival of B. bovis.