Fig. 3

Effects of pre-injection with constructs on the persistence of the anti-Clec9A-Ag constructs in the serum and on Ab responses. a, b Serum persistence. C57BL/6 mice were injected i.v. with a 2 μg rat anti-Clec9A-M2e mAb construct or b 2 μg rat anti-Clec9A-SP70 mAb construct, both constructs being made using the rat mAb clone 10B4. At day 28 after priming, all groups of mice were given a second injection of 1 μg anti-Clec9A-M2e or 1 μg anti-Clec9A-SP70, respectively. Serum samples were collected at the indicated times post immunisation and the concentration of the rat Ig mAb in the serum was measured by ELISA. Open circles represent the values from the primary response, closed circles the values from the boost response. Each point represents pooled mean with SEM from five mice. c Ab responses following pre-injection with anti-Clec9A mAb only. BALB/c mice (n = 5 mice per group) were injected i.v. with 100 μl PBS for the primary response group or with 2 μg anti-Clec9A mAb (clone 10B4, not linked to M2e) for the Ab pre-injected group. Two weeks later, both groups of mice were given an injection of 2 μg anti-Clec9A-M2e mAb construct. A further 2 weeks later, serum samples were collected and the anti-M2e IgG response was measured by ELISA. Each point represents one individual mouse and the end point titre is shown as geometric mean with 95% CI. Results represent a single experiment. Data were analysed by an unpaired two-tailed Student's t-test, and significance was indicated as *p < 0.05