HIGH-RESOLUTION MAPPING OF B-CELL EPITOPES WITHIN AN ANTIGENIC SEQUENCE FROM EIMERIA-TENELLA

Overlapping hexapeptides representing part of an Eimeria tenella antigenic sequence, shown to induce partial immunity to homologous challenge in chickens, were synthesized on polypropylene pins (Pepskan technique; Cambridge Research Biochemicals, Cambridge, United Kingdom). The binding to these hexapeptides of antibodies from chickens infected and rabbits immunized with five species of Eimeria was studied, using the coated pins as the solid phase of an enzyme-linked immunoassay. Antibody binding to most regions of the sequence was demonstrated, with peak areas of antigenicity correlating with the most hydrophilic regions. A particularly hydrophilic and antigenic area towards the N terminus of the sequence consists of a peptide motif repeated five times in the native antigen. Homologous antisera (chicken and rabbit anti-E. tenella antisera) differed in their pattern of reactivity from heterologous sera raised against other Eimeria species. While the former bound to fewer of the hexapeptides than the latter, they did so very strongly, indicating affinity maturation of the antibody response to E. tenella-specific sequences. No antibody reactivity to two regions of the sequence was detected. These regions occur in relatively hydrophilic areas and so are unlikely to be situated in transmembrane domains or in the interior of globular proteins. Synthetic peptides, as used in these experiments, make possible analysis of the fine specificity of immune responses and thus have a role to play in the development of novel vaccines for the control of coccidiosis.