Gene Expression Profile Of Actived Cytokine-induced Killer Cells

Objective: To investigate the stimulatory effects of DC on antitumor effects ofCIK and the changes of DC-induced CIK cell expression profile.Methods: DCs were generated by culture of adherent cells from PBMC for 7daysin the presence of IL-4 and GM-CSE CIKs were generated by culture ofnon-adherent cells for 7 or 14 days in the presence of INF-γ, IL-2, IL-1a, andmouse anti-human CD3 monoclonal antibody. The phenotype of DC and CIK wasanalyzed by FCM. The activity of DC was evaluated by MLR, and cytotoxicity ofCIK was assayed by MTT. We detected the CIK expression profile of cytokineand signal transduction genes by the DNA microarray under differentcircumstances.Results: Phenotypic analysis indicated that CD3~+ CD56~+ NKT cells werepredominant in the CIK cells generated in the current study, and that DC cellsexpressed high levels of CD40,CD80,CD86和HLA-DR. As demonstrated byMTT assay, the CIK cells proliferated rapidly, and the DC cells were able toinduce an MLR. Both antigen-pulsed and unpulsed DC stimulated theproliferation of CIK cells, and no significant difference was found between thetwo kinds of DC cells. Unpulsed DC cells did not enhance cytot0xicity mediatedby 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 beating tumors. InDNA microarray experiment, it was found 50 kinds of genes to be great changebeyond 5 folds.Conclusion: One of the major effects of antigen-pulsed DC is to activated CIKcells and to make them specific in killing of tumor cells. The changes ofDC-induced CIK expression profile are showed in Cys-Cys, MIP-2a, IFNregulatory factor-1,IFN-gamma antagonist cytokine,TGF-β,IL-5,FAST kinase andSARP3.