Chemokine fusion improves protein up take in vivo and in vitro. (A) hMIP3αsFvLF, MIP3αsFv38, or sFvLF fusion proteins at 2 different concentrations (1 μg/mL and 0.1 μg/mL) were incubated for 4 hours with the patient's PBMC-derived DCs. Then, DCs were thoroughly washed, irradiated, and mixed with the patient-derived Id-specific T-cell lines. After 72 hours, IFN-γ, GM-CSF, and TNF-α production was assayed in culture supernatants. Control DCs were incubated with 100 μg/mL and 1 μg/mL of the patient B-cell lymphoma-derived IgM (Id-LF) or LPS (10 ng/mL). Representative of 2 experiments performed in duplicate wells. (B) BALB/c mice (3/group) were immunized subcutaneously with 25 μg endotoxin-free MIP3αVL315, DF2βsFv315, or sFv315 each, or mock injected with PBS. After 10 and 48 hours, lymph node (LN) cells had been produced, irradiated (2000 rad), and mixed directly (without any additional protein stimulation) with 7A10B2 T cells in 96-well round-bottom plates at a 1:1 ratio (2 × 104 cells each) for 48 hours. IFN-γ production was assessed by ELISA in culture supernatants. Representative of 2 consecutive experiments performed in duplicate wells are shown. Error bars depict standard error of the mean of 3 mice per group.