FIG. 1.

Fractalkine induction in endothelial cell cultures associated with CMV antigen-stimulated PBMC from CMV-seropositive donors. The data are representative of three separate experiments with different blood donors. (A) Detection of fractalkine induction in endothelial cell monolayers cocultured with CMV-seropositive PBMC by Western blot analysis with fractalkine-specific antibody. Lanes in left gel panel: 1, positive control for fractalkine, with ∼85-kDa form of fractalkine (note that the fractalkine form [R&D] lacks 57 carboxy-terminal amino acids and migrates at ∼85 kDa, whereas the endothelial-cell-associated form [also from R&D] is full length and migrates at ∼100 kDa); 2, negative control for fractalkine with endothelial cells in a resting state; 3 and 4, replicate samples of CMV antigen-treated PBMC from a CMV-seropositive donor; 5 and 6, replicate samples of media alone-treated PBMC from a CMV-seropositive donor. Lanes in right gel panel: 1, positive control for fractalkine, with ∼85-kDa form of fractalkine (commercially available from R&D); 2, negative control for fractalkine with endothelial cells in a resting state; 3 and 4, replicate samples of CMV antigen-treated PBMC from a CMV-seronegative donor; 5 and 6, replicate samples of medium-alone-treated PBMC from a CMV-seronegative donor. The arrow at the right of the gel indicates the M_r of fractalkine. (B) Detection of secreted fractalkine levels in coculture supernatants. ELISA results from coculture supernatants, comparing CMV-seropositive PBMC to CMV-seronegative PBMC.