【摘要】目的探讨小鼠骨髓树突状细胞(dendritic cell,DCs)， 体外 经Hepa 1-6肝癌细胞株抗原粗提物致敏后，对特异性细胞毒T淋巴细胞(CTL)的诱导作用。 方法自小鼠骨髓分离DCs前体细胞，经GM-CSF+IL-4培养、扩增； 制备H epa 1-6 小鼠肝癌细胞株抗原粗提物，体外脉冲DCs，检测DCs诱导同基因型小鼠T细胞 增殖能 力及特异性CTL的反应能力。结果和结论经肿瘤抗原致敏的DCs组 织相容 性分子(MHC-Ⅰ、Ⅱ)及共刺激分子(B7-2、B7-1)表达明显增高，能诱导较 强的同基因型小鼠T细胞增殖，且能诱导Hepa 1-6特异性细胞毒T淋巴细胞。

【中图分类号】R73-36⁺2【文献标识码】A【文章编号】0529-1356(200 0)增-89

In vitro INDUCTION OF ANTI-TUMOR RESPONSIVENESS BY DENDRITIC CELLS PULSED WITH TUMOR EXTRACTS

WANG Guo-qiang,ZHONG Cui-ping,FAN Qiang,FU Ji-dong,GU Yung-di

(1.Deptartment of Histology and Embryology;)

HUANG Hui

(Department of Immmunol ogy;)

YE Sheng-long

(Liver Cancer Institute，Shanghai Medical University,Shanghai 200032,Chi na)

【Abstract】Objective The present stud y was designed to inv estigate whether pulsing DCs with tumor-derived extracts is an effective way to induce CTL and antitumor immunity.Methods DCs were propagated f r om bone marrow (BM)of C57BL/6J(H-2K^b、I-A^b)mice in vitro with GM -CSF+IL- 4tumor associated antigen (TAA)extracted from actively growing Hepa 1-6 cells w as used to activate DCs.The phenotypes of DCs were detected by FACS,the cytotoxi c ity of CTL was assayed by ³H-TdR labbel assay.Result and Conclusi on The TAA extract pulsed DCs exhibited much more and longer cell processe s and increased expression of MHC-I,MHC-II,CD80(B7-1)、CD86(B7-2).This expe riment has shown that DCs pulsed with TAA extracts of C57B/6J cells could stimul ate effectively the responsiveness of syngenic splenic T cells to induce specifi c CTL against C57BL/6J cells.

【Key word】Dendritic cells;Hepatocarcinoma;Antigen;Cytoto xic of T-lymphocytes

【基金项目】国家自然科学基金资助项目(39770387)

【作者简介】王国强(1970-)，男(汉族)，山东省博兴县人，在读博士研究 生

参考文献

［1］曹雪涛.树突状细胞与肿瘤的免疫治疗和基因治疗［J］.中国肿瘤生物治疗 杂志，1998，5(2)：82～84.

［2］Bandchereav J,Steinman RM.Dendritic cells and the control of immun ity［J］. Nature,1998,392(19):245251.

［3］Flamand.V,Sornasse T,Thielemans K,et al.Murine dendritic cells pul sed in vitro with tumor antigen induce tumor resistance in vivo［J］.Europ J Immunol,19 94 24(3):605-610.

［4］Nair SK,Snyder D,Rouse BT,et al.Regression of tumors in mice vacci nated w ith professional antigen-presenting cells pulsed with tumor extracts［J］.Int J Cancer,1997,70(6):706-715.

［5］沈关心，周汝麟.现代免疫学实验技术［M］.武汉：湖北科学技术出版社，1 998：179～187，390～392，301～303.

［6］Inaba K,Inaba M,Romani N,et al.Generation of large numbers of dendritic c ells from mouse bone marrow cultures supplemented with GM-CSF［J］. J Exp Med,1 992 176(6):16931702.

［7］吴厚生，谢琪，姜小玲，等.用³H-TdR释放法测量细胞介导的细胞毒功能 ［J］.上海免疫学杂志，1987，7(4)：230～235.

［8］Viola A.Lanzavecchia A T cell activation determined by Tcell recep tor number and tunable thresholds［J］. Science,1996,273(5271):104-106.

［9］刘彬彬，叶胜龙，贺平，等.肝癌患者外周血树突状细胞的体外扩增、鉴定 及吞噬能力观察［J］.肿瘤，1999，19(5)：259262.

【收稿日期】2000-07-04【修回日期】2000-08-02