Immunity to Eimeria is species-specific, and chickens with immunity to one species of Eimeria remain susceptible to other Eimeria species. This presents a major challenge in the development of effective vaccines against multiple Eimeria species. In this study, we cloned the antigenic epitope of a tachyzoite surface protein gene of Eimeria tenella, a tachyzoite surface protein gene of Eimeria acervulina and the gametocyte protein gene of Eimeria maxima, and constructed prokaryotic and eukaryotic plasmids carrying the multi-epitope antigenic gene. Immunization of chickens with the multivalent DNA and protein conferred partial protection against infection by the three Eimeria species, as shown by increased CD4+ T lymphocytes in the intestinal mucosa, decreased oocyst excretion and intestinal lesions, and increased body weight gain compared with non-immunized controls. The DNA prime-protein boost immunization schedule induced greater cellular immunity and protection from Eimeria infection than immunization with DNA or protein alone. Our findings demonstrated that DNA prime-protein boost immunization with a multivalent vaccine could stimulate protective immunity against challenge infection of multiple Eimeria species. This work provides a promising step towards DNA-protein vaccination against multiple species of pathogens.
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