Figure 6

Detection of Tax-specific CTLs in primary splenocytes stimulated with FPM1.BP cells in vitro (A) Splenocytes were isolated from an HTLV-I infected (▪) or uninfected control (□) rat and then stimulated with formalin-fixed FPM1.BP cells twice with 1-week interval. One week after the first or second stimulation, splenocytes were purified and then incubated with the 293T cells transfected with pEF/RT1AlSCNLEnv371L-EGFP or pEF/RT1AlSCTax180L-EGFP. One hour after the mixed culture, cells were stained with PE-conjugated anti-rat CD8 antibody and EGFP expression on CD8+ cells were assessed by flow cytometric analysis. The percent CD8+ cells that stain positively with each SCT-EGFP were shown. The data represent the mean ± the SD of triplicate analyses. (B) One week after the first or second stimulation, splenocytes were purified and then stimulated with 10 μM of Tax180-188 or NLEnv371-379 peptides for 48 hours. Production of IFN-γ in the culture supernatants was measured by ELISA. The data represent the mean ± the SD of triplicate wells.