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. 2005 Jun;79(12):7544–7557. doi: 10.1128/JVI.79.12.7544-7557.2005

FIG. 8.

FIG. 8.

Newly synthesized SV5 proteins are readily detected in significant quantities in C57BL/6 BMDC but not in BALB/c BMDC following infection with rSV5. BMDC from C57BL/6 (A) or BALB/c (D) mice were cultured in vitro for 7 days as described in Materials and Methods. On day 7, cells were mock infected (M) or infected with rSV5-wt or rSV5-GFP (MOI = 10). At 24 h postinfection, cells were stained with a primary hamster anti-mouse antibody to CD11c, followed by the secondary antibody (goat anti-hamster Alexa-568) and immunofluorescence was analyzed. The images shown are representative of three independent experiments. P815 cells, C57BL/6 BMDC (B), or BALB/c BMDC (E) were either mock infected (M) or infected with live or UV-inactivated rSV5 (MOI = 10). Two and 24 hours after infection, cells were lysed and analyzed by Western blotting for SV5 P protein and cellular actin. The blot shown is representative of three independent experiments. Note: The amount of SV5-infected P815 lysate used was 15 μg, whereas 30 μg of the other samples were loaded. (C) C57BL/6 BMDC were infected on day 7 with live or UV-inactivated rSV5-GFP-Ova (MOI = 10). Twenty-four hours after infection, C57BL/6 BMDC were cocultured with naive OT-1 TCR transgenic CD8+ T cells (specific for Ova257-264 peptide) at a ratio of DC:T cells of 1:10. Twenty-four hours following coculture, IFN-γ present in the supernatant was measured by ELISA. (F) BALB/c BMDC were infected on day 7 with live or UV-inactivated rSV5-I10 (MOI = 10). At 24 h postinfection, BMDC were cocultured with an I10-specific high avidity CTL line at a ratio of DC:T cells of 1:10. Twenty-four hours following coculture, IFN-γ present in the supernatant was measured by ELISA. The graphs show the average ± standard error of the mean of three independent experiments. P values were calculated using the paired t test. *, a significant decrease (P = 0.029) in the amount of IFN-γ produced by CD8+ cocultured with C57BL/6 BMDC infected with UV-inactivated rSV5-GFPOva relative to IFN-γ produced by CD8+ T cells cultured with BMDC infected with live rSV5-GFPOva.