The immunoregulatory activity of a nonmammalian interleukin 2 (IL-2), chicken IL-2 (chIL-2), was investigated using a DNA vaccine against infectious bursal disease virus (IBDV) as a model. Coadministration of a plasmid encoding the VP2 gene of IBDV (pCI-VP2) and a plasmid encoding chicken IL-2 gene (pCI-chIL-2) enhances bursal protection against both the homologous IBDV strain ZJ2000 and the heterologous strain BC6/85 compared to administration of pCI-VP2 alone. Vaccination with pCI-VP2 alone induces low bursal protection against ZJ2000 and only protects chickens from clinical outbreaks and mortality, but not from bursal damage caused by BC6/85. Co-administration of the plasmid encoding the polyprotein gene of IBDV (pCI-VP2/4/3) and pCI-chIL-2 provides complete protection (15/15) against ZJ2000 and satisfactory protection (13/15) against BC6/85. In contrast, only 10 out of 15 chickens and 6 out of 15 chickens were protected against ZJ2000 and BC6/85, respectively, using the pCI-VP2/4/3 vaccination alone. A significant increase in the IBDV-specific neutralizing antibody response was also observed in chickens that received pCI-VP2/4/3 plus pCI-chIL-2 as compared with those that received the pCI-VP2/4/3 vaccination alone. By administrating different amounts of plasmid DNA, we confirmed that the pCI-chIL-2, but not the backbone plasmid pCI, contributes to increased immunoprotection of DNA vaccine against IBDV. These results strongly indicate that the efficacy of avian DNA vaccine can be modulated by co-administration of a plasmid encoding chIL-2.