Fig 6. Tolerance and cross-tolerance to splenocyte cytokine release is unaffected by blocking PD-1.

The presented data was derived from at least 3 independent experiments, each in triplicate. Data represents splenocyte cytokine release 48 hours after successive 24-hour exposures to the ligand treatment combinations detailed on the X-axis. (A) Pre-treatment (30 minutes) of Flt3L-expanded splenocytes (1 × 10⁶ cells/ml) with an mouse anti-PD-1 blocking antibody (or corresponding isotypic control antibody, PD-1C) (5 μg/ml) prior to the secondary 24-hour ligand treatment had no effect upon resultant TNFα release regardless of the treatment combination employed. (B) Similarly, anti-PD-1 pre-treatment did not alter resultant splenocyte IFNγ release for any of the treatment combinations used. (C) Consistently, resultant splenocyte IL-10 release was unaffected for all treatment combinations by anti-PD-1 pre-treatment.