Potentiation of antigen-specific, Th1 immune responses by multiple DNA vaccination with an ovalbumin/interferon-γ hybrid construct

The preferential differentiation of T helper (Th) cells to Th1 or Th2 subsets is important with respect to susceptibility or resistance to particular infections, or to autoimmune diseases and allergic diseases. To more effectively drive immune responses toward antigen-specific Thl responses, we constructed a mammalian expression vector (pOVA/IFN-γ) carrying a hybrid gene in which the ovalbumin (OVA) (a model antigen) cDNA was covalently linked to murine interferon-γ (IFN-yγ) cDNA. Intramuscular injection of BALB/c mice with the pOVA/IFN-γ DNA increased both the production of OVA-specific IFN-γ by CD4⁺ T cells and the ratio of anti-OVA immunoglobulin G (IgG)2a to IgG1 isotypes, while the injection with the pOVA alone, or with the mixture of the pOVA and pIFN-γ, caused no or little increase. Furthermore, the OVA-specific, Thl immune responses were dramatically augmented by multiple injections with the pOVA/IFN-γ DNA. These studies indicate that the direct linkage of an OVA gene to an IFN-γ gene in the expression plasmid is required for efficiently confining the Thl effects of IFN-γ to the OVA-specific cells, and the linkage effect of the OVA/IFN- γ DNA can be potentiated by multiple vaccination.