Cytokine-induced Killer Cell Activation by Dentritic Cells Loaded with the Tumor Antigen from Human Lung Cancer Cell Line A549 / 中国慢性病预防与控制

ABSTRACT

Objective To explore the specific anti-tumor effect of cytokine-induced killer cell (CIK) activation by dentritic cells (DC) loaded with the tumor antigen from human lung cancer cell line A549. Methods CIKs and DCs derived from the health donor's peripheral blood monocyte (PBMC) were isolated and induced by the cytokine in vitro. DCs were generated by culture of adherent cells from PBMC for 7days in the presence of IL-4 and GM-CSF. CIKs were generated by culture of non-adherent cells for 7 or 14 days in the presence of INF-?, IL-2, IL-1?, and mouse anti-human CD3 monoclonal antibody. The phenotype of DC and CIK was analyzed by FCM. The activity of DC was evaluated by MLR; and the cytotoxicity of CIK was assayed by MTT. The CIK expression profile of cytokine and signal transduction genes were detected by the DNA micro-array under different circumstances. Results Phenotypic analysis indicated that CD3+CD56+NKT cells were predominant in the CIK cells generated in the current study, and that DC cells expressed high levels of CD40, CD80, CD86 and HLA-DR. As demonstrated by MTT assay, the CIK cells proliferated rapidly, and the DC cells were able to induce an MLR. Both antigen-pulsed and unpulsed DC stimulated the proliferation of CIK cells, and no significant difference was found between the two kinds of DC cells. Unpulsed DC cells did not enhance cytotoxicity mediated by CIK cells even though they were able to stimulate CIK proliferation. Antigen-pulsed DC, however, stimulated CIK cells to specifically kill target cells. The specific antitumor effect was also observed in nude mice bearing tumors. Conclusion One of the major effects of antigen-pulsed DC is to activate CIK cells and to make them specific in killing tumor cells.