Abstract:

[Abstract] Objective  To investigate the destructive effect of CSC-DC-CIK who were induced by cytokine induced killer (CIK) cells co-cultured with dendritic cells (DCs) on homologous tumor cells and to explore the possibility of CSC anti? gen involving in killing tumor. Methods Kidney cancer stem cells (KSCs) and lung cancer stem cells (LSCs) were isolated through FACS using CD133 + as a selection marker from cultured kidney cancer cell line A498 and lung cancer cell line A549 respectively. Freeze-thaw method was used to obtain the cancer stem cells（CSCs）antigens. DC cells and CIK cells were collected by in vitro expansion and inducted from the mononuclear cells isolated from human cord blood. The CIK cells were co-cultured with the DCs which were pulsed with the CSCs antigens（CSC-DC-CIK）mentioned above. Immunopheno? types of DC and CIK were analyzed by flow cytometry; cytokines levels were detected by ELISA kits and the destructive ef? fects of two kinds of CSC-DC-CIKs were tested by lactate dehydrogenase (LDH) release assay. Results  The expression of phenotypes CD40+, CD80+, CD86+ and HLA-DR+ were higher in CSC-DC than in CD（P < 0.01）; the expression of pheno? types CD40+, CD80+, CD86+ and HLA-DR+ of DC and CSC-DC were higher after co-culture than those before co-culture（ P < 0.01）;the expression of phenotypes CD40+, CD80+, CD86+ and HLA-DR+ of CSC-DC after been co-cultured with CIK were higher than those of DC after been co-cultured with CIK（P < 0.01）. The CIK phenotypes: CD3+, CD8+, CD56+ were in? creased in CIK co-cultured with both CSC-DC and DC than those before co-culture (P < 0.01); the expression of pheno? types CD3+, CD8+, CD56 +were higher in CSC-DC co-cultured with CIK than in DC co-cultured with CIK. DC-CIK and CSC-DC-CIK groups were more capable to express IFN-γ, TNF-α, IL-2 than they were before co-cultured with CIK (P < 0.01). CSC-DC-CIK group can secrete more above cytokines than DC-CIK group does（P < 0.01）. The destructive rates of KSC-DC-CIK and LSC-DC-CIK on target cells were (50.21±4.24)% and (49.32±3.89)% respectively which were much higher than that in DC-CIK（30.25±3.11）%（F = 89.157，P < 0.01）. Conclusion CSC-DC-CIKs have destructive effects on homologous tumor cells. More researches are needed to explore the mechanism and to evaluate the clinical applications.

Key words: Immunohistochemistry, dendritic cells, immunotherapy, cytokine induced killer