荷CEA-rV的DC诱导的CIK对CEA阳性原代肿瘤细胞的特异杀伤作用

目的：探讨携带CEA重组基因痘苗病毒（CEA-rV）抗原的树突状细胞（DC）诱导的杀伤细胞（CIK）对CEA阳性原代肿瘤细胞的特异性杀伤作用。方法：用脐血制备脐血单个核细胞（UBMC）、DC、CIK、DC＋CIK、CEArV＋DC＋CIK和CEA-rV＋DC＋UNBC细胞。取新鲜切除的食管癌组织和肺癌组织，制备原代培养的CEA阳性肿瘤细胞作为靶细胞。用MTT法测5组效应细胞对两种靶细胞的杀伤活性。结果：1）用CEA兔抗人单克隆抗体鉴定证实两种原代培养靶细胞系CEA阳性细胞。2）流式细胞仪测定DC的分子表型，证实为DC。3）流式细胞仪测定CIK的CD3^＋CD56^＋双阳性细胞达75．4％。4）DC＋CIK和CIK相比，对两种CEA阳性肿瘤细胞的杀伤率差异无统计学意义，P〉0．05。5）CE-rV4-DC＋CIK对两种CEA阳性肿瘤细胞的杀伤率（78．80％）明显高于DC＋CIK（52．94％），P％0．05。结论：CEA—rV＋DC能明显提高CIK细胞对原代培养的CEA阳性肿瘤细胞的杀伤活性，而单纯DC不能提高CIK的杀伤活性，说明CEA—rV在提高CIK对CEA阳性肿瘤细胞的特异杀伤作用中起重要作用。

Special killing effect of CIK induced by DC loaded with CEA-rV on CEA-positive primary tumor cells

OBJECTIVE:To survey the special killing activity of cytokine induced killer cells(CIK) induced by dendritic cells (DC) loaded with CEA recombinant vaccinia virus(CEA-rV) on CEA-positive tumor cells of primary culture. METHODS: Umbilical blood mononuclear cells(UBMC), DC, CIK, DC+CIK, CEA-rV+DC+CIK, and CEA-rV+DC+UMBC were prepared from umbilical blood. The CEA-positive primary tumor cells derived from freshly esophageal carcinoma tissues and lung cancer tissues were the target cell groups. The killing activity of each effector cell, was measured respectively by MTT reduction assay. RESULTS: 1) Two primary tumor cells confirmed by CEA monoclonal antibody of rabbit anti-human were really CEA positive cells. 2)It was showed by flow-cytometry that the mature DC expressed molecule such as CD86, CD83, CD80 and CD40 highly. 3)There were 75.4% double positive cells expressing the CD3/CD56 in CIK cells by flow-cytometry. 4)For comparing the killing activity to 2 CEA-positive primary tumor cells of CIK group and DC+CIK group, there was no significant difference in statistics, P>0.05. 5)The killing activity to 2 CEA-positive primary tumor cells of CEA-rV+DC+CIK was 78.80%, which was higher than that of DC+CIK about 52.94%. So CEA-rV could distinctly promote the special killing activity to CEA positive tumor cells of CIK, P<0.05. CONCLUSIONS: CEA-rV+DC could obviously enhance the special killing activity of CIK to CEA-positive primary tumor cells, while the DC alone could not. CEA-rV plays an important role during the special killing activity of CIK cells to CEA positive tumor cells.