Genetically Modified Lactococcus lactis for Delivery of Human Interleukin-10 to Dendritic Cells
(a) Immature DC were matured with no bacteria, 1 × 105 cfu of viable L. lactisIL-10 or L. lactis in the presence of MF. Mature DC were subsequently cultured with naïve Th-cells to induce effector Th-cells which were harvested, washed, and stained with 3 × 10−5 M PKH-26, a red cell-cycle tracking dye. Then, 2.5 × 104 effector Th-cells were preactivated overnight with anti-CD3 (1 : 5000) and anti-CD28 (0.5 mg/mL) in round-bottom 96-well plates. The following day allogenic (CD4+) Th-cells were labeled with CFSE, a green cell-cycle tracking dye. After 5 days, the cellular content of CFSE in the allogenic Th-cells was analyzed by flow cytometry . Depicted is the cell cycle progression of allogenic Th-cells stimulated in the presence of MF derived effector Th-cells (white histograms, reference condition) and of allogenic Th-cells cultured in the presence of MF/L. lactisIL-10 or MF/L. lactis derived effector Th-cells (gray histograms). (b) Depicted is the mean inhibition of fluorescence intensity of allogenic Th-cells from five separate experiments for the indicated conditions. The proliferation of allogenic Th-cells cultured in the presence of effector Th-cells derived from MF-matured DC which is regarded as 100% (reference condition). Mean percentages with standard error of mean are given; *indicates and **indicates using the -test for comparing percentages with the reference condition (MF).
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