Culture of Dendritic Cell from Normal Peripheral Blood Monocyte and Its Anti-tumor Immune Activity When Pulsed by Renal Cell Carcinoma Cell Line: In vitro Study / 대한비뇨기과학회지

ABSTRACT

PURPOSE: Dendritic cells (DC) play a crucial role in the initiation of primary immune response and are known as an excellent adjuvant for anti-cancer immunity. In this study, we tried to obtain substantial numbers of DC from peripheral blood of normal volunteers. We also investigated the anti-tumor immune response of DC pulsed by renal cell carcinoma cell line A498 in vitro. MATERIALS AND METHODS: DC were generated by culturing plastic adherent mononuclear cells from the peripheral blood in the presence of granulocyte-macrophage colony- stimulating factor and interleukin-4. Immature DC were cocultured with T-cells and pulsed by A498. MTT analysis was performed using the medium in which A498 only was cultured as control. Our experiments were analyzed by means of a commercial IL-12 p70 ELISA (Quantikine; R & D Systems, Minneapolis, MN). The capture antibodies used in both tests specifically recognize the p70 heterodimer, but not the free p40 chains. Detection limits were 30pg/ml of IL-12. RESULTS: We could obtain 1.5-2.0x106 DC with phenotype typical of mature DC (CD14-, CD80+, and CD83+) from the normal peripheral blood. On T-cell proliferation assay, the number of T-cells increased in proportion to that of DC and when DC were pulsed by A498, the same phenomenon could be observed. DC and T-cell media with A498 tumor lysate showed more production of IL-12 on IL-12 p70 ELISA than the media without A498 tumor lysate. CONCLUSIONS: We could successfully obtain mature DC from the peripheral blood. The data revealed indirectly that DC treated with tumor lysate enhance immune activity and thereby increase the anti-cancer effect of T-cells. Further investigations including in-vivo study are necessary to realize the effect of immunotherapy using DC against metastatic renal cell carcinoma.