Inhibitory effect of interferon gamma on frequency of Ehrlichia canis-infected cells in vitro

Ehrlichia canis is an obligate intracellular bacterium that infects the macrophage–monocyte cells of dogs, causing canine monocytic ehrlichiosis. Interferon-γ (IFN-γ), along with other cytokines, mediates the immune response to such intracellular bacterial invasions. To determine the role of IFN-γ in the immunity of dogs to E. canis infection, peripheral blood mononuclear cells (PBMC) and white blood cells (WBC) were collected from E. canis-infected dogs and added to a culture of E. canis in DH82 cells. The number of E. canis inclusion-positive cells was significantly reduced in cultures containing PBMC and WBC from E. canis-infected dogs compared to uninfected dogs. However, this resistance was inhibited by the addition of an anti-dog IFN-γ antibody. Resistance was also observed when PBMC were added to the Cell Culture Inserts, which prohibited contact of PBMC to DH82 cells, while allowed the diffusion of soluble cell products. The results of this study indicate that resistance was not dependent on cell to cell contact, but was associated with soluble cell products, such as IFN-γ. The addition of recombinant canine IFN-γ to the E. canis culture also reduced the number of infected cells. A commercial recombinant canine IFN-γ, which is sold in Japan, was also effective at reducing E. canis-infected cell number. These results indicate that IFN-γ has an inhibitory effect on the frequency of E. canis-infected cells in vitro and that contact between effector and target cells is not necessary for the resistance.