Fig. 5

Cytokine mRNA expression and enhanced IFN-γ and IL-2 production in T cells treated with anti-4-1BB scFvs. a Following 72 h of treatment with anti-4-1BB scFvs, cells were collected and analyzed for IFN-γ, IL-2 and IL-4 mRNA expression levels using real-time PCR, with the amplification normalized against HPRT. Results from quantitative real-time PCR demonstrate that mRNA for IFN-γ and IL-2 was significantly increased in T cells treated with PI.12, PII.16 and PII.29 scFvs.PI.42 and that ES1 scFvs could not induce IFN-γ and IL-2 production in T cells. No IL-4 mRNA expression was observed in the treated cells. NS: no significant difference. *p < 0.05. Data represent the mean of three experiments ± SD. b Cytokine secretion by T-cells treated with anti-4-1BB scFvs was determined using ELISA. IFN-γ and IL-2 increased significantly following treatment of T cells with PI.12, PII.16 and PII.29 scFvs (p < 0.05), while the increase in IL-4 production was not significant. The results shown are means ± SD amounts of cytokine secreted by treated T cells (in pg/ml). Each experiment was repeated a minimum of three times