小鼠肝癌抗原粗提物致敏树突状细胞体外诱导特异性细胞毒T淋巴细胞的研究
作者：王国强 钟翠平 范强 傅继东 顾云娣 黄辉 叶胜龙
单位：王国强 钟翠平 范强 傅继东 顾云娣(上海医科大学组织学胚胎学教研室)；黄辉(免疫学教研室)；叶胜龙(肝癌研究所，上 海200032)
关键词：树突状细胞；肝癌；抗原；细胞毒T淋巴细胞
解剖学报00zk23
【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。
, http://www.100md.com
【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;)
, 百拇医药
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-2Kb、I-Ab)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.
, http://www.100md.com
【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):245251.
, 百拇医药
[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.
, http://www.100md.com
[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):16931702.
[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)：259262.
【收稿日期】2000-07-04 【修回日期】2000-08-02, http://www.100md.com
单位：王国强 钟翠平 范强 傅继东 顾云娣(上海医科大学组织学胚胎学教研室)；黄辉(免疫学教研室)；叶胜龙(肝癌研究所，上 海200032)
关键词：树突状细胞；肝癌；抗原；细胞毒T淋巴细胞
解剖学报00zk23
【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。
, http://www.100md.com
【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;)
, 百拇医药
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-2Kb、I-Ab)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.
, http://www.100md.com
【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):245251.
, 百拇医药
[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.
, http://www.100md.com
[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):16931702.
[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)：259262.
【收稿日期】2000-07-04 【修回日期】2000-08-02, http://www.100md.com