The understanding of innate immune modulation by pathogens and immune-modulating agents, including synthetic oligodeoxynucleotides (CpG ODNs), has offered several new approaches to improve prophylactic and therapeutic strategies against infectious diseases in humans and animals. However, in this regard not much work has been done in avian medicine. In the present study, we analyzed the kinetics of interferon (IFN), cytokine, and chemokine mRNA expression in chicken embryonic spleen at 6 hr, 24 hr, 48 hr, and 72 hr after administration of CpG ODN 2007 (B-class) in 18-day-old chicken embryos. Our data showed enhanced expression of IFN-γ; interleukin (IL)-1β, IL-6, and IL-8; and oligoadenyl synthetase A mRNA after CpG ODN administration. In addition, CpG ODN administration to chicken embryos 24 hr before the challenge with infectious bronchitis virus (IBV) was capable of limiting IBV propagation in different embryonic tissues. Based on the kinetics and type of cytokines induced after in ovo administration of CpG ODN, it may be speculated that in ovo administration of CpG ODNs may enhance resistance from viral infection in neonatal chicks and that CpG ODNs may contribute toward the development of more effective and safer poultry vaccines including in ovo vaccines.