Interleukin-10 (IL-10) plays an indispensable role in mucosal tolerance by programming dendritic cells (DCs) to induce suppressor Th-cells. We have tested the modulating effect of L. lactis secreting human IL-10 ( 𝐿 . 𝑙 𝑎 𝑐 𝑡 𝑖 𝑠 𝐼 𝐿 - 1 0 ) on DC function in vitro. Monocyte-derived DC incubated with 𝐿 . 𝑙 𝑎 𝑐 𝑡 𝑖 𝑠 𝐼 𝐿 - 1 0 induced effector Th-cells that markedly suppressed the proliferation of allogenic Th-cells as compared to L. lactis. This suppressive effect was only seen when DC showed increased CD83 and CD86 expression. Furthermore, enhanced production of IL-10 was measured in both 𝐿 . 𝑙 𝑎 𝑐 𝑡 𝑖 𝑠 𝐼 𝐿 - 1 0 -derived DC and Th-cells compared to L. lactis-derived DC and Th-cells. Neutralizing IL-10 during DC-Th-cell interaction and coculturing 𝐿 . 𝑙 𝑎 𝑐 𝑡 𝑖 𝑠 𝐼 𝐿 - 1 0 -derived suppressor Th-cells with allogenic Th-cells in a transwell system prevented the induction of suppressor Th-cells. Only 130 pg/mL of bacterial-derived IL-10 and 40 times more exogenously added recombinant human IL-10 were needed during DC priming for the generation of suppressor Th-cells. The spatially restricted delivery of IL-10 by food-grade bacteria is a promising strategy to induce suppressor Th-cells in vivo and to treat inflammatory diseases.