The Cultivation Of Dendritic Cells(DC) And DC Transfected With Carcinoembryonic Antigen-vaccinia Recombinant Virus Induces CEA-specific Immunity Mediated By Cytotoxic T Lymphocytes In Vitro

hi our study,dendritic cells (DCs) were derived from the cultivation of peripheral blood monocytes in vitro successfully. Then to observe whether DCs transfected with carcinoembryonic antigen (CEA)-vaccinia recombinant virus (rV-CEA) can induces cytotoxic T lymphocyte-mediated CEA-specific immunity in vitro.First,peripheral blood monocytes from patients of carcinoma were treated with rhGM-CSF of 1000 U/ml plus IL-4 of 500 U/ml (A group) rhGM-CSF of 1000 U/ml (B group) and simply RPMI 1640 (C group). After cultivation of 7 days,change of cellular morphology was observed under light microscopy and electron microscopy. Surface markers on DCs were then analyzed by flow cytometry and the proliferation of T cells was detected by MTT colorimetry. Resoults:Peripheral blood monocytes from patients of carcinoma treated with rhGM-CSF of 1000 U/ml plus IL-4 of 500 U/ml for 7 days could observe DCs with typical morphology. Simultaneously there was a decrease in CD 14 expression and increase in HLA-DR,CD40,CD83 and CD86 on DCs. In addition,it could stimulate evidently proliferation of T cells. Conclusion:dendritic cells(DCs) can be derived from peripheral blood monocytes in vitro successfully treated with rhGM-CSF of 1000 U/ml plus IL-4 of 500 U/ml.Second,DCs from the patients of advanced carcinoma of large intestine were transfected with rV-CEA,and then cocultured with autologous T cells. The cytotoxic activity against CEA-secreting tumor cells were assessed and compared with those of T cells induced by DCs without rV-CEA transfection. Results:DCs transduced with rV-CEA are effective stimuli of T lymphocyte proliferation. T cells induced by DCs transfected with rV-CEA showed specific cytotoxicity against CEA-secreting tumor cells.