Figure 2.

Absence of recall proliferation in CD4⁺ T cells from FVIII plasmid plus anti-CD3–treated mice. CD4⁺ T cells were isolated by MACS from spleens of naive, anti-CD3 only, FVIII plasmid only, and FVIII plasmid + anti-CD3–treated mice 3 weeks after plasmid injection. A total of 1.0 × 10⁵ CD4⁺ T cells were cocultured with irradiated 1.0 × 10⁵ CD4⁻ cells in 96-well round-bottom plates with or without the presence of FVIII at 10 U/mL for 72 hours, followed by adding 1 μCi of [³H]thymidine per well for the final 18 hours. *P < .05, compared with the group of FVIII plasmid only. Data shown are mean ± SD of counts per minute of [³H]thymidine incorporation in triplicate wells.