Development and characterization of chicken-chicken B cell hybridomas secreting monoclonal antibodies that detect sporozoite and merozoite antigens of Eimeria.

Chicken-chicken B cell hybridomas that secrete monoclonal antibodies (mAb) detecting antigens located at the anterior tip of the sporozoites of Eimeria acervulina have been produced. Peripheral blood lymphocytes (PBL) obtained from chickens infected with E. acervulina were fused with a thymidine-kinase deficient (TK-) chicken B cell line. Hybridomas secreting mAb binding to the sporozoite antigens of E. acervulina were selected using an ELISA. Two stable hybridomas, whose mAb were designated as P11 and P66, respectively, were obtained and their binding characteristics were assessed. The mAb P11 detects the sporozoite but not the merozoite antigens of E. acervulina and Eimeria tenella. The P66 mAb recognizes a cross-reactive antigen of sporozoite and merozoite of E. acervulina but not E. tenella. Flow cytometric analysis of the hybridomas secreting the P11 and P66 mAb showed expression of surface IgM and IgG, respectively. Western blot analysis under reducing conditions showed that mAb P11 recognized a sporozoite protein of 43 kDa. Both mAb recognized antigens located at the anterior end of the sporozoites by indirect immunofluorescence staining. This study shows the development of chicken B cell hybridomas that secrete coccidia-specific mAb.