树突状细胞诱导供体脾淋巴细胞过继性免疫治疗预防肝移植术后肝癌复发的实验研究
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摘要
第一部分 大鼠肝移植急性排斥反应模型的建立
目的:掌握大鼠肝移植模型的制作方法,结合国内外报道,对大鼠肝移植模型进行技术改进,并观察以SD大鼠为供体Wistar大鼠为受体时免疫排斥反应发生的情况,建立稳定的大鼠肝移植急性排斥反应模型。
材料和方法:应用Kamada“二袖套法”行大鼠肝移植,在既往国内外文献报道的基础上对供肝切取、灌注、袖套管制作、受体麻醉、肝上下腔静脉吻合和缩短无肝期等方面进行改进,熟练实验操作。行SD→wistar大鼠肝移植20例,并随机分为排斥反应组、环孢霉素A处理组(CsA,5mg/kg)各10例,以wistar→Wistar同系移植组10例作为对照组,观察移植术后免疫排斥情况。
结果:供体手术时间29.6±5.2min,供肝修整时间20.1±3.3min,受体总手术时间45.7±5.9min,受体无肝期17.2±2.4min,肝上下腔静脉缝合时间10.5±2.2min,手术成功率93.3%。排斥反应组大鼠术后逐渐出现黄疸、腹水等表现,肝功能进行性恶化、最终出现肝功能衰竭症状而死亡,生存期5～15天,组织病理学提示有典型的免疫排斥,而对照组和CsA治疗组大鼠均存活1月以上。术后第7天排斥反应组血清IFN-γ、IL-4水平显著升高,而应用CsA治疗组血清IFN-γ和IL-4水平处于低水平。
结论:对大鼠肝移植技术进行改良可明显提高移植大鼠的生存率。以SD大鼠
Part 1 The establishment of acute rejection model after allogenic rat orthotopic liver transplantationObjective To study and reform the technique of rat orthotopic liver transplantation (ROLT), and observe the allograft rejection of liver transplantation for SD→Wistar rat.Methods ROLTs were performed according to Kamada's two cuff technique with some modifications in donor liver acquirement, perfusion, cuff made, recipient anesthesia, anastomosis of suprahepatic inferior vena cava (SHIVC) and shorten anhepatic time. 20 cases who underwent SD→Wistar ROLT were randomly divided into two groups, rejection group and cyclosporine treatment group(CsA group), with 10 cases each. 10 cases who underwent Wistar→Wistar syngenic ROLT were taken for control group. Posttransplantation rejection were observed in each group. Results The time used for total donor operation was 29. 6 ± 5. 2min,
trimming of donor liver 20.1+3. 3min, total recipient operation 45.7 ±5.9min, anhepatic phase 17.2 + 2. 4min, anastomosis of SHIVC 10.5 + 2. 2min. More than 93. 3% of the operations were successful. Jaundice, ascites, progressively deterioration of liver function, and finally liver failure could be observed in rats of the rejection group with survival time 5-15 days after allogenic liver transplantation. All of the rats of the control group and CsA group survived more than 1 month. Serum IFN-yand IL-4 level was significantly elevated in the rejection group on the 7th day after transplantation. Both serum IFN- y and IL-4 were at low level in CsA group.Conclusion Posttransplantion survival rate was significantly increased after the reform of surgical technique. The donor liver of SD rats and recipient Wistar rats combination is a high rejection model, and it can be effectively used to study allograft rejection. IFN- y is one of the most important cytokine of liver allograft rejection. As an good index for indication of rejection at early phase of transplantion, high concentration of serum IFN- Y is correlated with the severity of rejection. Activation of T cells and their differentiation from ThO to Thl and Th2 were suppressed by use of CsA, as well as the allograft rejection, but tolerance of the allograft was not induced.Part 2 Recurrence of hepatocellular carcinomar in livergraft: an animal model
Objective To explore a new way to develop a rat model of tumor recurrence in liver allograft with intrahepatic implantation of tumor fragments and the use of immunosuppressant, mimic the progress of clinical hepatocellular carcinoma (HCC) recurrence in liver graft, study tumor recurrence and tumor growth in liver grafts in recipients with HCC under well-defined conditions.Methods CBRH-7919 cell line, which is syngenic to Wistar rat, were cultured in vivo, then implanted subcutaneously in the Wistar rats and serially passaged prior to implantation in the liver. Small tumor fragments of 1X1X lmm were implanted under the liver capsule in the left lateral or median liver lobe. The tumor size was measured with ultrasound under ether anesthesia after 7 days and the tumor volume was calculated. Group A (n = 5), a control group, a tumor was implanted and measured in Wistar livers. In group B (n = 5), a SD-*Wistar allogenic ROLT was performed and a fresh tumor piece was implanted after reperfusion. In group C (n = 5), a tumor was implanted in the liver 7 days after ROLT. Tumor growth curves were compared.Results Tumor take was five of five in group A, one of five in group B, and five of five in group C. A significant growth retardation of the tumor was seen in group B as compared to controls. No difference in growth rate was observed between group C and controls.Conclusion Tumor implantation in the allograft liver 7 days after ROLT under immunosuppression established a safe and reliable rat model of tumor recurrence in liver graft.
Part 3 Experimental study on the prevention forpost-transplantation recurrence of hepatocel lular carcinomawith dendritic cells induced donor spleen lymphocytes invitro and in vivoObjective To study the feasibility and validity of adoptive immunotherapy using dendritic cell(DC) activated tumor specific lymphocytes from donor spleen cells to prevent recipient hepatocellular carcinoma(HCC) recurrence after liver transplantation, and to develop an effective method to eliminate residue cancer cells of the recipient, breakthrough the tolerance state to cancer and decrease the recurrence rate of the liver graft.Methods The precursors of dendritic cells were isolated from bone marrow of recipient rats(Wistar), stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4) and cultured for 5 days. The DCs were identified with microscopic morphology and phenotype. Then rat DCs were pulsed with CBRH-7919 tumor cell antigens prepared with freeze-thawed tumor cell lysates. Spleen cells from donor(SD)/recipient(Wistar) rat were cocultured with these DCs respectively to induce CTL as two experimental groups, and unactivited blank donor/recipient spleen cells were
maintained survival by lower dose of rrIL-2 as two control groups. Supernatant IFN- y release were determined by ELISA, and the cytotoxicity of CTL was assayed by LDH release test. 26 cases of posttransplantation HCC recurrence rat model were randomly selected into five groups, 6 cases each. Four groups were treated with adoptive lymphocyte infusion which come from the correspondent groups in vitro, and another group was saline infusion group for blank control. Tumorgenecity, tumor infiltrating-lymphocyte, tumor growth, rat survival, hepatic impairment, rejection, side effect and serum cytokine level were compared between each group.Results Spleen cells activated by DCs loaded with tumor antigen manifested much higher cytotoxic to tumor cells than unactivated group and saline control group. The cytotoxisity of activated cells from donor spleen was significantly higher than that from the resipient. Cytotoxisity to the tumor cells was not observed in both donor and recipient orgin unactivated lymphocytes group. The aggregation of lymphocytes within and near the tumor tissue with necrosis of tumor cells, retardation of the tumor growth, decrease of the tumorgenecity and prolong of the survival time were observed in activated donor spleen cell infusion group without any evidence of liver toxicity and side effect. Some tumor suppression were also observed in activated recipient origin spleen cell infusion group, such as prolong of the survival time and tumor growth retardation, but serum aminotransaminase enzyme was significantly elevated. Conclusion Donor spleen cells infusion which activated by DCs pulsed
with tumor antigen is an practical, safe and effective way to prevent and treat posttransplantation recurrence of HCC. It can decrease postoperative recurrence rate, suppress tumor growth and prolong survival time without any impairment to the liver graft, and it has better treatment and prevention effect than the spleen cells from recipient.
目的:掌握大鼠肝移植模型的制作方法,结合国内外报道,对大鼠肝移植模型进行技术改进,并观察以SD大鼠为供体Wistar大鼠为受体时免疫排斥反应发生的情况,建立稳定的大鼠肝移植急性排斥反应模型。
材料和方法:应用Kamada“二袖套法”行大鼠肝移植,在既往国内外文献报道的基础上对供肝切取、灌注、袖套管制作、受体麻醉、肝上下腔静脉吻合和缩短无肝期等方面进行改进,熟练实验操作。行SD→wistar大鼠肝移植20例,并随机分为排斥反应组、环孢霉素A处理组(CsA,5mg/kg)各10例,以wistar→Wistar同系移植组10例作为对照组,观察移植术后免疫排斥情况。
结果:供体手术时间29.6±5.2min,供肝修整时间20.1±3.3min,受体总手术时间45.7±5.9min,受体无肝期17.2±2.4min,肝上下腔静脉缝合时间10.5±2.2min,手术成功率93.3%。排斥反应组大鼠术后逐渐出现黄疸、腹水等表现,肝功能进行性恶化、最终出现肝功能衰竭症状而死亡,生存期5～15天,组织病理学提示有典型的免疫排斥,而对照组和CsA治疗组大鼠均存活1月以上。术后第7天排斥反应组血清IFN-γ、IL-4水平显著升高,而应用CsA治疗组血清IFN-γ和IL-4水平处于低水平。
结论:对大鼠肝移植技术进行改良可明显提高移植大鼠的生存率。以SD大鼠
Part 1 The establishment of acute rejection model after allogenic rat orthotopic liver transplantationObjective To study and reform the technique of rat orthotopic liver transplantation (ROLT), and observe the allograft rejection of liver transplantation for SD→Wistar rat.Methods ROLTs were performed according to Kamada's two cuff technique with some modifications in donor liver acquirement, perfusion, cuff made, recipient anesthesia, anastomosis of suprahepatic inferior vena cava (SHIVC) and shorten anhepatic time. 20 cases who underwent SD→Wistar ROLT were randomly divided into two groups, rejection group and cyclosporine treatment group(CsA group), with 10 cases each. 10 cases who underwent Wistar→Wistar syngenic ROLT were taken for control group. Posttransplantation rejection were observed in each group. Results The time used for total donor operation was 29. 6 ± 5. 2min,
trimming of donor liver 20.1+3. 3min, total recipient operation 45.7 ±5.9min, anhepatic phase 17.2 + 2. 4min, anastomosis of SHIVC 10.5 + 2. 2min. More than 93. 3% of the operations were successful. Jaundice, ascites, progressively deterioration of liver function, and finally liver failure could be observed in rats of the rejection group with survival time 5-15 days after allogenic liver transplantation. All of the rats of the control group and CsA group survived more than 1 month. Serum IFN-yand IL-4 level was significantly elevated in the rejection group on the 7th day after transplantation. Both serum IFN- y and IL-4 were at low level in CsA group.Conclusion Posttransplantion survival rate was significantly increased after the reform of surgical technique. The donor liver of SD rats and recipient Wistar rats combination is a high rejection model, and it can be effectively used to study allograft rejection. IFN- y is one of the most important cytokine of liver allograft rejection. As an good index for indication of rejection at early phase of transplantion, high concentration of serum IFN- Y is correlated with the severity of rejection. Activation of T cells and their differentiation from ThO to Thl and Th2 were suppressed by use of CsA, as well as the allograft rejection, but tolerance of the allograft was not induced.Part 2 Recurrence of hepatocellular carcinomar in livergraft: an animal model
Objective To explore a new way to develop a rat model of tumor recurrence in liver allograft with intrahepatic implantation of tumor fragments and the use of immunosuppressant, mimic the progress of clinical hepatocellular carcinoma (HCC) recurrence in liver graft, study tumor recurrence and tumor growth in liver grafts in recipients with HCC under well-defined conditions.Methods CBRH-7919 cell line, which is syngenic to Wistar rat, were cultured in vivo, then implanted subcutaneously in the Wistar rats and serially passaged prior to implantation in the liver. Small tumor fragments of 1X1X lmm were implanted under the liver capsule in the left lateral or median liver lobe. The tumor size was measured with ultrasound under ether anesthesia after 7 days and the tumor volume was calculated. Group A (n = 5), a control group, a tumor was implanted and measured in Wistar livers. In group B (n = 5), a SD-*Wistar allogenic ROLT was performed and a fresh tumor piece was implanted after reperfusion. In group C (n = 5), a tumor was implanted in the liver 7 days after ROLT. Tumor growth curves were compared.Results Tumor take was five of five in group A, one of five in group B, and five of five in group C. A significant growth retardation of the tumor was seen in group B as compared to controls. No difference in growth rate was observed between group C and controls.Conclusion Tumor implantation in the allograft liver 7 days after ROLT under immunosuppression established a safe and reliable rat model of tumor recurrence in liver graft.
Part 3 Experimental study on the prevention forpost-transplantation recurrence of hepatocel lular carcinomawith dendritic cells induced donor spleen lymphocytes invitro and in vivoObjective To study the feasibility and validity of adoptive immunotherapy using dendritic cell(DC) activated tumor specific lymphocytes from donor spleen cells to prevent recipient hepatocellular carcinoma(HCC) recurrence after liver transplantation, and to develop an effective method to eliminate residue cancer cells of the recipient, breakthrough the tolerance state to cancer and decrease the recurrence rate of the liver graft.Methods The precursors of dendritic cells were isolated from bone marrow of recipient rats(Wistar), stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4) and cultured for 5 days. The DCs were identified with microscopic morphology and phenotype. Then rat DCs were pulsed with CBRH-7919 tumor cell antigens prepared with freeze-thawed tumor cell lysates. Spleen cells from donor(SD)/recipient(Wistar) rat were cocultured with these DCs respectively to induce CTL as two experimental groups, and unactivited blank donor/recipient spleen cells were
maintained survival by lower dose of rrIL-2 as two control groups. Supernatant IFN- y release were determined by ELISA, and the cytotoxicity of CTL was assayed by LDH release test. 26 cases of posttransplantation HCC recurrence rat model were randomly selected into five groups, 6 cases each. Four groups were treated with adoptive lymphocyte infusion which come from the correspondent groups in vitro, and another group was saline infusion group for blank control. Tumorgenecity, tumor infiltrating-lymphocyte, tumor growth, rat survival, hepatic impairment, rejection, side effect and serum cytokine level were compared between each group.Results Spleen cells activated by DCs loaded with tumor antigen manifested much higher cytotoxic to tumor cells than unactivated group and saline control group. The cytotoxisity of activated cells from donor spleen was significantly higher than that from the resipient. Cytotoxisity to the tumor cells was not observed in both donor and recipient orgin unactivated lymphocytes group. The aggregation of lymphocytes within and near the tumor tissue with necrosis of tumor cells, retardation of the tumor growth, decrease of the tumorgenecity and prolong of the survival time were observed in activated donor spleen cell infusion group without any evidence of liver toxicity and side effect. Some tumor suppression were also observed in activated recipient origin spleen cell infusion group, such as prolong of the survival time and tumor growth retardation, but serum aminotransaminase enzyme was significantly elevated. Conclusion Donor spleen cells infusion which activated by DCs pulsed
with tumor antigen is an practical, safe and effective way to prevent and treat posttransplantation recurrence of HCC. It can decrease postoperative recurrence rate, suppress tumor growth and prolong survival time without any impairment to the liver graft, and it has better treatment and prevention effect than the spleen cells from recipient.
引文
1. Lee S, Charters AC, Chandler JG, et al. A technique for orthotopic liver transplantation in the rat. Transplantation, 1973, 16(6): 664-669.
2. Komatsu T, Yamauchi K, Furukawa T, et al. Transcatheter arterial injection of autologous lymphokine-activated killer (LAK) cells into patients with liver cancers. J Clin Immunol, 1990, 10(3): 167-174.
3. Haruta I, Yamauchi K, Aruga A, et al. Analytical study of the clinical response to two distinct adoptive immunotherapies for advanced hepatocellular carcinoma: comparison between LAK cell and CTL therapy. J Immunother Emphasis Tumor Immunol, 1996, 19(3): 218-223.
4. Yamarnoto M, Fujii H. Optimal locoregional immunochemotherapy after tumor-mass reduction for advanced hepatoeellular carcinoma. Hepatogastroenterology, 1995, 42(5): 567-577.
5. Oka M, Hazama S, Yoshino S, et al. Intraarterial combined immunochemotherapy for unresectable hepatocellular carcinoma: preliminary results. Cancer Immunol Immunother, 1994, 38(3): 194-200.
6. Kamada N, Calne RY. A surgical experience with five hundred thirty liver transplants in the rat. Surgery, 1983, 93(1 Pt 1): 64-69.
7. Williams JW, Peters TG, Vera SR, et al. Biopsy-directed immunosuppression following hepatic transplantation in man. Transplantation, 1985, 39(6): 589-596.
8. Kamada N, Calne RY. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage. Transplantation, 1979, 28(1): 47-50.
9. Miyata M, Fischer JH, Fuhs M, et al. A simple method for orthotopic liver transplantation in the rat. Cuff technique for three vascular anastomoses. Transplantation, 1980, 30(5): 335-338.
10. Engemann R, Ulrichs K, Thiede A, et al. Value of a physiological liver transplant model in rats. Induction of specific graft tolerance in a fully allogeneic strain combination. Transplantation, 1982, 33(5): 566-568.
11. Tsuchimoto S, Kusumoto K, Nakajima Y, et al. Orthotopic liver transplantation in the rat. A simplified technique using the cuff method for suprahepatie vena cava anastomosis. Transplantation, 1988, 45(6): 1153-1155.
12. Harihara Y, Sanjo K, Idezuki Y. A modified cuff technique for suprahepatic vena cava anastomosis in rat liver transplantation. Transplantation, 1992, 53(3): 707-709.
13.张绍庚,方石岗.改良三袖套法大鼠原位肝移植.中华显微外科杂志,1997,2054-55.
14.王轩,杨甲梅,严以群,姚晓平,吴孟超.大鼠原位肝移植不同术式的探讨.中华器官移植杂志,1998,19(2):76-78.
15. Lee S, Charters AC, 3rd, OrloffMJ. Simplified technic for orthotopic liver transplantation in the rat. Am J Surg, 1975, 130(1): 38-40.
16.马毅,何晓顺,陈规划,陈细桃,黄洁夫.重建肝动脉血供大鼠原位肝移植模型的术式探讨.中华实验外科杂志,2004,21(1):110-111.
17.彭勇,龚建平,刘长安,王炜,严律南.大鼠原位肝移植模型制作过程中麻醉方法的选择.中国普通外科杂志,2003,12(9):673-676.
18. Haas O, Calmus Y, Legendre C, et al. Transplantation of allogeneic isolated hepatocytes induces a specific prolongation of allograft survival. Transplant Proc, 1990, 22(5): 2247-2249.
19. Zimmermann FA, Butcher GW, Davies HS, et al. Techniques for orthotopic liver transplantation in the rat and some studies of the immunologic responses to fully allogeneic liver grafts. Transplant Proc, 1979, 11 (1): 571-577.
20.许赤,黄洁夫,陈规划,何晓顺.大鼠肝移植术后早期急性排斥反应与细胞因子基因表达的关系.肝胆外科杂志,2000,8(3):229-230.
21.王学浩,孙倍成,钱建民,张峰,李相成.转FasL基因的树突状细胞诱导大鼠肝移植免疫耐受的实验研究.中华肝胆外科杂志,2002,8(5):294-296.
22.章爱斌,郑树森,贾长库,王雁.骨化三醇预防大鼠原位肝移植后急性排斥的动态观察.中华普通外科杂志,2003,18(12):138-740.
23.姜明山,韩德恩.不同品系大鼠之间原位肝移植的实验观察.中华实验外科杂志,2004,21(4):128-129.
24.刘静,高毅,汪爽,孙尔维,张志,单毓强,钟世镇.引起排斥反应模型肝移植大鼠生存时间延长分析.广东医学,2004,25(12):1374-1377.
25. Neuberger J. Liver allograft rejection—current concepts on diagnosis and treatment. J Hepatol, 1995, 23 Suppl 154-61.
26. Mosmann TR, Cherwinskki H, Bond MW, et al. Two types of routine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol, 1986, 136(7): 2348-2357.
27. Wang YL, Tang ZQ, Gao W, et al. Influence ofThl, Th2, and Th3 cytoldnes during the early phase after liver transplantation. Transplant Proc, 2003, 35(8): 3024-3025.
28. Fujimoto N, Ishida H, Nakamura I, et al. Quantities of interleukin-12p40 in mature CD8alpha negative dendritic cells correlate with strength of TCR signal and determine Th cell development. Microbiol Immunol, 2003, 47(12): 1017-1024.
29. Wu S, Gessner R, von Stackelberg A, et al. Cytokine/cytokine receptor gene expression in childhood acute lymphoblastic leukemia: correlation of expression and clinical outcome at first disease recurrence. Cancer, 2005, 103(5): 1054-1063.
30. Curry H, Alvarez GR, Zwilling BS, et al. Toll-like receptor 2 stimulation decreases IFN-gamma receptor expression in mouse RAW264. 7 macrophages. J Interferon Cytokine Res, 2004, 24(12): 699-710.
31. Wells AD, Li XC, Li Y, et al. Requirement for T-cell apoptosis in the induction of peripheral transplantation tolerance. Nat Med, 1999, 5(11): 1303-1307.
32. Konieczny BT, Dai Z, Elwood ET, et al. IFN-gamma is critical for long-term allograft survival induced by blocking the CD28 and CD40 ligand T cell costimulation pathways. J Immunol, 1998, 160(5): 2059-2064.
33. Dai Z, Konieczny BT, Baddoura FK, et al. Impaired alloantigen-mediated T cell apoptosis and failure to induce long-term allograit survival in IL-2-deficient mice. J Immunol, 1998, 161(4): 1659-1663.
34. Huang WH, Yah Y, De Boer B, et al. A short course of cyclosporine immunosuppression inhibits rejection but not tolerance of rat liver allogratts. Transplantation, 2003, 75(3): 368-374.
35. Hemming AW, Cattral MS, Reed AI, et al. Liver transplantation for hepatocellular carcinoma. Ann Surg, 2001, 233(5): 652-659.
36.李齐根,杨广顺,卫立辛,杨宁,周学平,吴孟超.大鼠肝癌肝移植模型的建立.中华肝胆外科杂志,2004 10(3):191-195.
37.曾兆林,韩德恩,韩丽姝,汪鹰扬,王志东,张新宇.三氧化二砷预防大鼠肝癌肝移植后肿瘤复发的研究.中华实验外科杂志,2005,22(2):169-170.
38. Kienle P, Weitz J, Klaes R, et al. Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma. Arch Surg, 2000, 135(2): 213-218.
39. Carlsson G, Gullberg B, Hafstrom L. Estimation of liver tumor volume using different formulas-an experimental study in rats. J Cancer Res Clin Oncol, 1983, 105(1): 20-23.
40. Bruix J. Treatment ofhepatocellular carcinoma. Hepatology, 1997, 25(2): 259-262.
41. Ringe B, Wittekind C, Bechstein WO, et al. The role of liver transplantation in hepatobiliary malignancy. A retrospective analysis of 95 patients with particular regard to tumor stage and recurrence. Ann Surg, 1989, 209(1): 88-98.
42. Bismuth H, Chiche L, Adam R, et al. Liver resection versus transplantation for hepatocellular carcinoma in cirrhotic patients. Ann Surg, 1993, 218(2): 145-151.
43. Mazzaferro V, Regalia E, Doci R, et al. Liver transplantation for the treatment of small hepatocellular carcinomas in patients with cirrhosis. N Engl J Med, 1996, 334(11): 693-699.
44. Durand F, Belghiti J. Liver transplantation for hepatocellular carcinoma: should we push the limits? Liver Transpl, 2003, 9(7): 697-699.
45. Roberts JP, Liu T, Freise CE, et al. Liver transplantation improves survival of rats bearing hepatoma-3924A. J Surg Res, 1996, 65(1): 59-62.
46. Yang R, Rescorla FJ, Reilly CR, et al. A reproducible rat liver cancer model for experimental therapy: introducing a technique of intrahepatic tumor implantation. J Surg Res, 1992, 52(3): 193-198.
47. Matsuzaki T, Murase N, Yagihashi A, et al. Liver transplantation for diethylnitrosamine-induced hepatoeellular carcinoma in rats. Transplant Proc, 1992, 24(2): 748-751.
48. Yano K, Fukuda Y, Sumimoto R, et al. Development of a rat model for orthotopic liver transplantation for hepatocellular carcinoma. Surgery, 1995, 118(3): 539-546.
49. Freise CE, Ferrell L, Liu T, et al. Effect of systemic cyclosporine on tumor recurrence after liver transplantation in a model of hepatocellular carcinoma. Transplantation, 1999, 67(4): 510-513.
50. Travis CC, MeClain TW, Birkner PD. Diethylnitrosamine-induced hepatocarcinogenesis in rats: a theoretical study. Toxicol Appl Pharmacol, 1991, 109(2): 289-304.
51. Schotman SN, van Ijken MG, Marquet RL, et al. Tumor recurrence in liver grafts: two animal models. Transplant Proc, 1998, 30(4): 1052-1053.
52.王锦波,何振平.移植法建立大鼠肝癌模型.实验动物科学与管理,1999,16(2):27-28.
53.朱德厚,叶秀珍,陈瑞铭.大鼠肝癌细胞(CBRH-7919)的建株及其生物学特性研究.实验生物学报,1980,13(1):113.117.
54.程文,张青萍,贡雪灏,瘳晓峰.超声监护下免疫抑制大鼠肝癌模型的建立.中国医学影像技术,2003,19(7):840-842.
55.庄志祥,刘根寿,周剑影,段莹莹,李军成,吴浩荣.树突状细胞治疗大鼠肝癌的实验研究.中华普通外科杂志,2001 16(4):228-230.
56. Schlitt HJ, Neipp M, Weimann A, et al. Recurrence patterns of hepatocellular and fibrolamellar carcinoma after liver transplantation. J Clin Oncol, 1999, 17(1): 324-331.
57.王德臣,宋世兵,袁炯,修典荣.朱建平,蒋斌,张同琳.肝移植治疗晚期原发性肝癌的初步报告.中华肿瘤杂志,2003,25(3):295-297.
58.杨冬华.肝癌的免疫治疗现状和前景.中华肝脏病杂志,2003,11(12):757-758.
59. Iwatsuki S, Starzl TE, Sheahan DG, et al. Hepatic resection versus transplantation for hepatoeellular carcinoma. Ann Surg, 1991, 214(3): 221-228;discussion 228-229.
60. Jonas S, Bechstein WO, Steinmuller T, et al. Vascular invasion and histopathologic grading determine outcome after liver transplantation for hepatocellular carcinoma in cirrhosis. Hepatology, 2001, 33(5): 1080-1086.
61. Yoo HY, Patt CH, Geschwind JF, et al. The outcome of liver transplantation in patients with hepatocellular carcinoma in the United States between 1988 and 2001: 5-year survival has improved significantly with time. J Clin Oneol, 2003, 21(23): 4329-4335.
62. Yao FY, Kinkhabwala M, LaBerge JM, et al. The impact of pre-operative loco-regional therapy on outcome after liver transplantation for hepatocellular carcinoma. Am J Transplant, 2005, 5(4 Pt 1): 795-804.
63. Roayaie S, Frischer JS, Emre SH, et al. Long-term results with multimodal adjuvant therapy and liver transplantation for the treatment of hepatocellular carcinomas larger than 5 centimeters. Ann Surg, 2002, 235(4): 533-539.
64. Vivarelli M, Bellusci R, Cucchetti A, et al. Low recurrence rate of hepatocellular carcinoma after liver transplantation: better patient selection or lower immunosuppression? Transplantation, 2002, 74(12): 1746-1751.
65. Carr BI, Selby R, Madariaga J, et al. Prolonged survival after liver transplantation and cancer chemotherapy for advanced-stage hepatocellular carcinoma. Transplant Proc, 1993, 25(1 Pt 2): 1128-1129.
66. Majno PE, Adam R, Bismuth H, et al. Influence of preoperative transarterial lipiodol chemoembolization on resection and transplantation for hepatocellular carcinoma in patients with cirrhosis. Ann Surg, 1997, 226(6): 688-701;discussion 701-683.
67. Yao FY, Ferrell L, Bass NM, et al. Liver transplantation for hepatocellular carcinoma: expansion of the tumor size limits does not adversely impact survival. Hepatology, 2001, 33(6): 1394-1403.
68.张同琳.肝移植对晚期肝癌治疗的价值.中华医学杂志2004,84(18):1499-1500.
69.李坚,王洪林.肝癌肝移植术后肝癌复发的研究进展.中华外科杂志,2005,43(11):753-756.
70.杨甲梅,温增庆.肝移植术治疗原发性肝癌.肝胆外科杂志,1999,7166-167.
71.朱继业,刘燕南.肝癌的免疫治疗.临床外科杂志,2001,9(1):7-9.
72. Austyn JM. The dendritic cell system and anti-tumour immunity. In Vivo, 1993, 7(3): 193-201.
73. Steinman RM, Cohn ZA. Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med, 1973, 137(5): 1142-1162.
74. Banchereau J, Steinman RM. Dendritic cells and the control of immunity. Nature, 1998, 392(6673): 245-252.
75. Steinman RM. The dendritic cell system and its role in immunogenicity. Annu Rev Immunol, 1991, 9271-296.
76. Mailliard RB, Dallal RM, Son YI, et al. Dendritic cells promote T-cell survival or death depending upon their maturation state and presentation of antigen. Immunol Invest, 2000, 29(2): 177-185.
77. Ninomiya T, Akbar SM, Masumoto T, et al. Dendritic cells with immature phenotype and defective function in the peripheral blood from patients with hepatocellular carcinoma. J Hepatol, 1999, 31(2): 323-331.
78. Nair SK, Hull S, Coleman D, et al. Induction of carcinoembryonic antigen (CEA)-specific cytotoxic T-lymphocyte responses in vitro using autologous dendritic cells loaded with CEA peptide or CEA RNA in patients with metastatic malignancies expressing CEA. Int J Cancer, 1999, 82(1): 121-124.
79. Santin AD, Bellone S, Ravaggi A, et al. Induction of ovarian tumor-specific CDS+ cytotoxic T lymphocytes by acid-eluted peptide-pulsed autologous dendritic cells. Obstet Gynecol, 2000, 96(3): 422-430.
80. Inaba K, Metlay JP, Crowley MT, et al. Dendritic cells as antigen presenting cells in vivo. Int Rev Immunol, 1990, 6(2-3): 197-206.
81. Chan L, Linsley PS, Hellstrom KE. Costimulation of T cells for tumor immunity. Immunol Today, 1993, 14(10): 483-486.
82. Porgador A, Gilboa E. Bone marrow-generated dendritic cells pulsed with a class I-restricted peptide are potent inducers of cytotoxic T lymphocytes. J Exp Med, 1995, 182(1): 255-260.
83. Zitvogel L, Angevin E, Tursz T. Dendritic cell-based immunotherapy of cancer. Ann Oncol, 2000, 11 Suppl 3199-205.
84. Friedl J, Stiff A, Paolini P, et al. Tumor antigen pulsed dendritic cells enhance the cytolytic activity of tumor infiltrating lymphocytes in human hepatocellular cancer. Cancer Biother Radiopharm, 2000, 15(5): 477-486.
85. Porgador A, Snyder D, Gilboa E. Induction of antitumor immunity using bone marrow-generated dendritic cells. J Immunol, 1996, 156(8): 2918-2926.
86. Ladhams A, Schmidt C, Sing G, et al. Treatment of non-resectable hepatocellular carcinoma with autologous tumor-pulsed dendritic cells. J Gastroenterol Hepatol, 2002, 17(8): 889-896.
87. Iwashita Y, Tahara K, Goto S, et al. A phase I study of autologous dendritic cell-based immunotherapy for patients with unresectable primary liver cancer. Cancer Immunol Immunother, 2003, 52(3): 155-161.
88. Homma S, Toda G, Gong J, et al. Preventive antitumor activity against hepatocellular carcinoma (HCC) induced by immunization with fusions of dendritic cells and HCC cells in mice. J Gastroenterol, 2001, 36(11): 764-771.
89. Tatsumi T, Takehara T, Kanto T, et al. Administration of intedeukin-12 enhances the therapeutic efficacy of dendritic cell-based tumor vaccines in mouse hepatocellular carcinoma. Cancer Res, 2001, 61(20): 7563-7567.
90.黄波,冯作化,张桂梅.Hsp70肿瘤抗原肽复合物修饰的DC疫苗体内外特异性抗瘤作用.中国免疫学杂志,2002,18(9):614-617.
91.郭建巍,蔡美英,魏大鹏.树突状细胞负载甲胎蛋白、细胞毒性T细胞表位肽后的免疫应答.中华肝脏病杂志,2002,10(3):178-180.
92. Lu J, Leng X, Peng J, et al. Induction of cytotoxic T lymphocytes from the peripheral blood of a hepatocellular carcinoma patient using melanoma antigen-1 (MAGE-1) peptide. Chin Med J (Engl), 2002, 115(7): 1002-1005.
93. Hsu FJ, Benike C, Fagnoni F, et al. Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med, 1996, 2(1): 52-58.
94. Nestle FO, Alijagic S, Gilliet M, et al. Vaccination of melanoma patients with peptide-or tumor lysate-pulsed dendritic cells. Nat Med, 1998, 4(3): 328-332.
95. Appelbaum FR. Haematopoietic cell transplantation as immunotherapy. Nature, 2001, 411(6835): 385-389.
96. Dahl AM, Beverley PC, Stauss HJ. A synthetic peptide derived from the tumor-associated protein mdm2 can stimulate autoreactive, high avidity cytotoxic T lymphocytes that recognize naturally processed protein. J Immunol, 1996, 157(1): 239-246.
97. Moscovitch M, Slavin S. Anti-tumor effects of allogeneic bone marrow transplantation in (NZB X NZW)F1 hybrids with spontaneous lymphosarcoma. J Immunol, 1984, 132(2): 997-1000.
98. Fabre JW. The allogeneic response and tumor immunity. Nat Med, 2001, 7(6): 649-652.
99. Strair RK, Schaar D, Medina D, et al. Antineoplastie effects of partially HLA-matched irradiated blood mononuclear cells in patients with renal cell carcinoma. J Clin Oncol, 2003, 21(20): 3785-3791.
100. Ertl HC, Xiang ZQ. Genetic immunization. Viral Immunol, 1996, 9(1): 1-9.
101. Takayama T, Sekine T, Makuuchi M, et al. Adoptive immunotherapy to lower postsurgical recurrence rates of hepatocellular carcinoma: a randomised trial. Lancet, 2000, 356(9232): 802-807.
102. Wagner U, Schlebusch H, Kohler S, et al. Immunological responses to the tumor-associated antigen CA125 in patients with advanced ovarian cancer induced by the murine monoclonal anti-idiotype vaccine ACA125. Hybridoma, 1997, 16(1): 33-40.
103. Talmor M, Mirza A, Turley S, et al. Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures. Eur J Immunol, 1998, 28(3): 811-817.
104. Winzler C, Rovere P, . Rescigno M, et al. Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures. J Exp Med, 1997, 185(2): 317-328.
105.陈向荣,石汉平.树突状细胞的分离纯化与鉴定.上海免疫学杂志,2001,21(2):122-125.
106. Brenan M, Puklavec M. The MRC OX-62 antigen: a useful marker in the purification of rat veiled cells with the biochemical properties of an integrin. J Exp Med, 1992, 175(6): 1457-1465.
2. Komatsu T, Yamauchi K, Furukawa T, et al. Transcatheter arterial injection of autologous lymphokine-activated killer (LAK) cells into patients with liver cancers. J Clin Immunol, 1990, 10(3): 167-174.
3. Haruta I, Yamauchi K, Aruga A, et al. Analytical study of the clinical response to two distinct adoptive immunotherapies for advanced hepatocellular carcinoma: comparison between LAK cell and CTL therapy. J Immunother Emphasis Tumor Immunol, 1996, 19(3): 218-223.
4. Yamarnoto M, Fujii H. Optimal locoregional immunochemotherapy after tumor-mass reduction for advanced hepatoeellular carcinoma. Hepatogastroenterology, 1995, 42(5): 567-577.
5. Oka M, Hazama S, Yoshino S, et al. Intraarterial combined immunochemotherapy for unresectable hepatocellular carcinoma: preliminary results. Cancer Immunol Immunother, 1994, 38(3): 194-200.
6. Kamada N, Calne RY. A surgical experience with five hundred thirty liver transplants in the rat. Surgery, 1983, 93(1 Pt 1): 64-69.
7. Williams JW, Peters TG, Vera SR, et al. Biopsy-directed immunosuppression following hepatic transplantation in man. Transplantation, 1985, 39(6): 589-596.
8. Kamada N, Calne RY. Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage. Transplantation, 1979, 28(1): 47-50.
9. Miyata M, Fischer JH, Fuhs M, et al. A simple method for orthotopic liver transplantation in the rat. Cuff technique for three vascular anastomoses. Transplantation, 1980, 30(5): 335-338.
10. Engemann R, Ulrichs K, Thiede A, et al. Value of a physiological liver transplant model in rats. Induction of specific graft tolerance in a fully allogeneic strain combination. Transplantation, 1982, 33(5): 566-568.
11. Tsuchimoto S, Kusumoto K, Nakajima Y, et al. Orthotopic liver transplantation in the rat. A simplified technique using the cuff method for suprahepatie vena cava anastomosis. Transplantation, 1988, 45(6): 1153-1155.
12. Harihara Y, Sanjo K, Idezuki Y. A modified cuff technique for suprahepatic vena cava anastomosis in rat liver transplantation. Transplantation, 1992, 53(3): 707-709.
13.张绍庚,方石岗.改良三袖套法大鼠原位肝移植.中华显微外科杂志,1997,2054-55.
14.王轩,杨甲梅,严以群,姚晓平,吴孟超.大鼠原位肝移植不同术式的探讨.中华器官移植杂志,1998,19(2):76-78.
15. Lee S, Charters AC, 3rd, OrloffMJ. Simplified technic for orthotopic liver transplantation in the rat. Am J Surg, 1975, 130(1): 38-40.
16.马毅,何晓顺,陈规划,陈细桃,黄洁夫.重建肝动脉血供大鼠原位肝移植模型的术式探讨.中华实验外科杂志,2004,21(1):110-111.
17.彭勇,龚建平,刘长安,王炜,严律南.大鼠原位肝移植模型制作过程中麻醉方法的选择.中国普通外科杂志,2003,12(9):673-676.
18. Haas O, Calmus Y, Legendre C, et al. Transplantation of allogeneic isolated hepatocytes induces a specific prolongation of allograft survival. Transplant Proc, 1990, 22(5): 2247-2249.
19. Zimmermann FA, Butcher GW, Davies HS, et al. Techniques for orthotopic liver transplantation in the rat and some studies of the immunologic responses to fully allogeneic liver grafts. Transplant Proc, 1979, 11 (1): 571-577.
20.许赤,黄洁夫,陈规划,何晓顺.大鼠肝移植术后早期急性排斥反应与细胞因子基因表达的关系.肝胆外科杂志,2000,8(3):229-230.
21.王学浩,孙倍成,钱建民,张峰,李相成.转FasL基因的树突状细胞诱导大鼠肝移植免疫耐受的实验研究.中华肝胆外科杂志,2002,8(5):294-296.
22.章爱斌,郑树森,贾长库,王雁.骨化三醇预防大鼠原位肝移植后急性排斥的动态观察.中华普通外科杂志,2003,18(12):138-740.
23.姜明山,韩德恩.不同品系大鼠之间原位肝移植的实验观察.中华实验外科杂志,2004,21(4):128-129.
24.刘静,高毅,汪爽,孙尔维,张志,单毓强,钟世镇.引起排斥反应模型肝移植大鼠生存时间延长分析.广东医学,2004,25(12):1374-1377.
25. Neuberger J. Liver allograft rejection—current concepts on diagnosis and treatment. J Hepatol, 1995, 23 Suppl 154-61.
26. Mosmann TR, Cherwinskki H, Bond MW, et al. Two types of routine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol, 1986, 136(7): 2348-2357.
27. Wang YL, Tang ZQ, Gao W, et al. Influence ofThl, Th2, and Th3 cytoldnes during the early phase after liver transplantation. Transplant Proc, 2003, 35(8): 3024-3025.
28. Fujimoto N, Ishida H, Nakamura I, et al. Quantities of interleukin-12p40 in mature CD8alpha negative dendritic cells correlate with strength of TCR signal and determine Th cell development. Microbiol Immunol, 2003, 47(12): 1017-1024.
29. Wu S, Gessner R, von Stackelberg A, et al. Cytokine/cytokine receptor gene expression in childhood acute lymphoblastic leukemia: correlation of expression and clinical outcome at first disease recurrence. Cancer, 2005, 103(5): 1054-1063.
30. Curry H, Alvarez GR, Zwilling BS, et al. Toll-like receptor 2 stimulation decreases IFN-gamma receptor expression in mouse RAW264. 7 macrophages. J Interferon Cytokine Res, 2004, 24(12): 699-710.
31. Wells AD, Li XC, Li Y, et al. Requirement for T-cell apoptosis in the induction of peripheral transplantation tolerance. Nat Med, 1999, 5(11): 1303-1307.
32. Konieczny BT, Dai Z, Elwood ET, et al. IFN-gamma is critical for long-term allograft survival induced by blocking the CD28 and CD40 ligand T cell costimulation pathways. J Immunol, 1998, 160(5): 2059-2064.
33. Dai Z, Konieczny BT, Baddoura FK, et al. Impaired alloantigen-mediated T cell apoptosis and failure to induce long-term allograit survival in IL-2-deficient mice. J Immunol, 1998, 161(4): 1659-1663.
34. Huang WH, Yah Y, De Boer B, et al. A short course of cyclosporine immunosuppression inhibits rejection but not tolerance of rat liver allogratts. Transplantation, 2003, 75(3): 368-374.
35. Hemming AW, Cattral MS, Reed AI, et al. Liver transplantation for hepatocellular carcinoma. Ann Surg, 2001, 233(5): 652-659.
36.李齐根,杨广顺,卫立辛,杨宁,周学平,吴孟超.大鼠肝癌肝移植模型的建立.中华肝胆外科杂志,2004 10(3):191-195.
37.曾兆林,韩德恩,韩丽姝,汪鹰扬,王志东,张新宇.三氧化二砷预防大鼠肝癌肝移植后肿瘤复发的研究.中华实验外科杂志,2005,22(2):169-170.
38. Kienle P, Weitz J, Klaes R, et al. Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma. Arch Surg, 2000, 135(2): 213-218.
39. Carlsson G, Gullberg B, Hafstrom L. Estimation of liver tumor volume using different formulas-an experimental study in rats. J Cancer Res Clin Oncol, 1983, 105(1): 20-23.
40. Bruix J. Treatment ofhepatocellular carcinoma. Hepatology, 1997, 25(2): 259-262.
41. Ringe B, Wittekind C, Bechstein WO, et al. The role of liver transplantation in hepatobiliary malignancy. A retrospective analysis of 95 patients with particular regard to tumor stage and recurrence. Ann Surg, 1989, 209(1): 88-98.
42. Bismuth H, Chiche L, Adam R, et al. Liver resection versus transplantation for hepatocellular carcinoma in cirrhotic patients. Ann Surg, 1993, 218(2): 145-151.
43. Mazzaferro V, Regalia E, Doci R, et al. Liver transplantation for the treatment of small hepatocellular carcinomas in patients with cirrhosis. N Engl J Med, 1996, 334(11): 693-699.
44. Durand F, Belghiti J. Liver transplantation for hepatocellular carcinoma: should we push the limits? Liver Transpl, 2003, 9(7): 697-699.
45. Roberts JP, Liu T, Freise CE, et al. Liver transplantation improves survival of rats bearing hepatoma-3924A. J Surg Res, 1996, 65(1): 59-62.
46. Yang R, Rescorla FJ, Reilly CR, et al. A reproducible rat liver cancer model for experimental therapy: introducing a technique of intrahepatic tumor implantation. J Surg Res, 1992, 52(3): 193-198.
47. Matsuzaki T, Murase N, Yagihashi A, et al. Liver transplantation for diethylnitrosamine-induced hepatoeellular carcinoma in rats. Transplant Proc, 1992, 24(2): 748-751.
48. Yano K, Fukuda Y, Sumimoto R, et al. Development of a rat model for orthotopic liver transplantation for hepatocellular carcinoma. Surgery, 1995, 118(3): 539-546.
49. Freise CE, Ferrell L, Liu T, et al. Effect of systemic cyclosporine on tumor recurrence after liver transplantation in a model of hepatocellular carcinoma. Transplantation, 1999, 67(4): 510-513.
50. Travis CC, MeClain TW, Birkner PD. Diethylnitrosamine-induced hepatocarcinogenesis in rats: a theoretical study. Toxicol Appl Pharmacol, 1991, 109(2): 289-304.
51. Schotman SN, van Ijken MG, Marquet RL, et al. Tumor recurrence in liver grafts: two animal models. Transplant Proc, 1998, 30(4): 1052-1053.
52.王锦波,何振平.移植法建立大鼠肝癌模型.实验动物科学与管理,1999,16(2):27-28.
53.朱德厚,叶秀珍,陈瑞铭.大鼠肝癌细胞(CBRH-7919)的建株及其生物学特性研究.实验生物学报,1980,13(1):113.117.
54.程文,张青萍,贡雪灏,瘳晓峰.超声监护下免疫抑制大鼠肝癌模型的建立.中国医学影像技术,2003,19(7):840-842.
55.庄志祥,刘根寿,周剑影,段莹莹,李军成,吴浩荣.树突状细胞治疗大鼠肝癌的实验研究.中华普通外科杂志,2001 16(4):228-230.
56. Schlitt HJ, Neipp M, Weimann A, et al. Recurrence patterns of hepatocellular and fibrolamellar carcinoma after liver transplantation. J Clin Oncol, 1999, 17(1): 324-331.
57.王德臣,宋世兵,袁炯,修典荣.朱建平,蒋斌,张同琳.肝移植治疗晚期原发性肝癌的初步报告.中华肿瘤杂志,2003,25(3):295-297.
58.杨冬华.肝癌的免疫治疗现状和前景.中华肝脏病杂志,2003,11(12):757-758.
59. Iwatsuki S, Starzl TE, Sheahan DG, et al. Hepatic resection versus transplantation for hepatoeellular carcinoma. Ann Surg, 1991, 214(3): 221-228;discussion 228-229.
60. Jonas S, Bechstein WO, Steinmuller T, et al. Vascular invasion and histopathologic grading determine outcome after liver transplantation for hepatocellular carcinoma in cirrhosis. Hepatology, 2001, 33(5): 1080-1086.
61. Yoo HY, Patt CH, Geschwind JF, et al. The outcome of liver transplantation in patients with hepatocellular carcinoma in the United States between 1988 and 2001: 5-year survival has improved significantly with time. J Clin Oneol, 2003, 21(23): 4329-4335.
62. Yao FY, Kinkhabwala M, LaBerge JM, et al. The impact of pre-operative loco-regional therapy on outcome after liver transplantation for hepatocellular carcinoma. Am J Transplant, 2005, 5(4 Pt 1): 795-804.
63. Roayaie S, Frischer JS, Emre SH, et al. Long-term results with multimodal adjuvant therapy and liver transplantation for the treatment of hepatocellular carcinomas larger than 5 centimeters. Ann Surg, 2002, 235(4): 533-539.
64. Vivarelli M, Bellusci R, Cucchetti A, et al. Low recurrence rate of hepatocellular carcinoma after liver transplantation: better patient selection or lower immunosuppression? Transplantation, 2002, 74(12): 1746-1751.
65. Carr BI, Selby R, Madariaga J, et al. Prolonged survival after liver transplantation and cancer chemotherapy for advanced-stage hepatocellular carcinoma. Transplant Proc, 1993, 25(1 Pt 2): 1128-1129.
66. Majno PE, Adam R, Bismuth H, et al. Influence of preoperative transarterial lipiodol chemoembolization on resection and transplantation for hepatocellular carcinoma in patients with cirrhosis. Ann Surg, 1997, 226(6): 688-701;discussion 701-683.
67. Yao FY, Ferrell L, Bass NM, et al. Liver transplantation for hepatocellular carcinoma: expansion of the tumor size limits does not adversely impact survival. Hepatology, 2001, 33(6): 1394-1403.
68.张同琳.肝移植对晚期肝癌治疗的价值.中华医学杂志2004,84(18):1499-1500.
69.李坚,王洪林.肝癌肝移植术后肝癌复发的研究进展.中华外科杂志,2005,43(11):753-756.
70.杨甲梅,温增庆.肝移植术治疗原发性肝癌.肝胆外科杂志,1999,7166-167.
71.朱继业,刘燕南.肝癌的免疫治疗.临床外科杂志,2001,9(1):7-9.
72. Austyn JM. The dendritic cell system and anti-tumour immunity. In Vivo, 1993, 7(3): 193-201.
73. Steinman RM, Cohn ZA. Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med, 1973, 137(5): 1142-1162.
74. Banchereau J, Steinman RM. Dendritic cells and the control of immunity. Nature, 1998, 392(6673): 245-252.
75. Steinman RM. The dendritic cell system and its role in immunogenicity. Annu Rev Immunol, 1991, 9271-296.
76. Mailliard RB, Dallal RM, Son YI, et al. Dendritic cells promote T-cell survival or death depending upon their maturation state and presentation of antigen. Immunol Invest, 2000, 29(2): 177-185.
77. Ninomiya T, Akbar SM, Masumoto T, et al. Dendritic cells with immature phenotype and defective function in the peripheral blood from patients with hepatocellular carcinoma. J Hepatol, 1999, 31(2): 323-331.
78. Nair SK, Hull S, Coleman D, et al. Induction of carcinoembryonic antigen (CEA)-specific cytotoxic T-lymphocyte responses in vitro using autologous dendritic cells loaded with CEA peptide or CEA RNA in patients with metastatic malignancies expressing CEA. Int J Cancer, 1999, 82(1): 121-124.
79. Santin AD, Bellone S, Ravaggi A, et al. Induction of ovarian tumor-specific CDS+ cytotoxic T lymphocytes by acid-eluted peptide-pulsed autologous dendritic cells. Obstet Gynecol, 2000, 96(3): 422-430.
80. Inaba K, Metlay JP, Crowley MT, et al. Dendritic cells as antigen presenting cells in vivo. Int Rev Immunol, 1990, 6(2-3): 197-206.
81. Chan L, Linsley PS, Hellstrom KE. Costimulation of T cells for tumor immunity. Immunol Today, 1993, 14(10): 483-486.
82. Porgador A, Gilboa E. Bone marrow-generated dendritic cells pulsed with a class I-restricted peptide are potent inducers of cytotoxic T lymphocytes. J Exp Med, 1995, 182(1): 255-260.
83. Zitvogel L, Angevin E, Tursz T. Dendritic cell-based immunotherapy of cancer. Ann Oncol, 2000, 11 Suppl 3199-205.
84. Friedl J, Stiff A, Paolini P, et al. Tumor antigen pulsed dendritic cells enhance the cytolytic activity of tumor infiltrating lymphocytes in human hepatocellular cancer. Cancer Biother Radiopharm, 2000, 15(5): 477-486.
85. Porgador A, Snyder D, Gilboa E. Induction of antitumor immunity using bone marrow-generated dendritic cells. J Immunol, 1996, 156(8): 2918-2926.
86. Ladhams A, Schmidt C, Sing G, et al. Treatment of non-resectable hepatocellular carcinoma with autologous tumor-pulsed dendritic cells. J Gastroenterol Hepatol, 2002, 17(8): 889-896.
87. Iwashita Y, Tahara K, Goto S, et al. A phase I study of autologous dendritic cell-based immunotherapy for patients with unresectable primary liver cancer. Cancer Immunol Immunother, 2003, 52(3): 155-161.
88. Homma S, Toda G, Gong J, et al. Preventive antitumor activity against hepatocellular carcinoma (HCC) induced by immunization with fusions of dendritic cells and HCC cells in mice. J Gastroenterol, 2001, 36(11): 764-771.
89. Tatsumi T, Takehara T, Kanto T, et al. Administration of intedeukin-12 enhances the therapeutic efficacy of dendritic cell-based tumor vaccines in mouse hepatocellular carcinoma. Cancer Res, 2001, 61(20): 7563-7567.
90.黄波,冯作化,张桂梅.Hsp70肿瘤抗原肽复合物修饰的DC疫苗体内外特异性抗瘤作用.中国免疫学杂志,2002,18(9):614-617.
91.郭建巍,蔡美英,魏大鹏.树突状细胞负载甲胎蛋白、细胞毒性T细胞表位肽后的免疫应答.中华肝脏病杂志,2002,10(3):178-180.
92. Lu J, Leng X, Peng J, et al. Induction of cytotoxic T lymphocytes from the peripheral blood of a hepatocellular carcinoma patient using melanoma antigen-1 (MAGE-1) peptide. Chin Med J (Engl), 2002, 115(7): 1002-1005.
93. Hsu FJ, Benike C, Fagnoni F, et al. Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med, 1996, 2(1): 52-58.
94. Nestle FO, Alijagic S, Gilliet M, et al. Vaccination of melanoma patients with peptide-or tumor lysate-pulsed dendritic cells. Nat Med, 1998, 4(3): 328-332.
95. Appelbaum FR. Haematopoietic cell transplantation as immunotherapy. Nature, 2001, 411(6835): 385-389.
96. Dahl AM, Beverley PC, Stauss HJ. A synthetic peptide derived from the tumor-associated protein mdm2 can stimulate autoreactive, high avidity cytotoxic T lymphocytes that recognize naturally processed protein. J Immunol, 1996, 157(1): 239-246.
97. Moscovitch M, Slavin S. Anti-tumor effects of allogeneic bone marrow transplantation in (NZB X NZW)F1 hybrids with spontaneous lymphosarcoma. J Immunol, 1984, 132(2): 997-1000.
98. Fabre JW. The allogeneic response and tumor immunity. Nat Med, 2001, 7(6): 649-652.
99. Strair RK, Schaar D, Medina D, et al. Antineoplastie effects of partially HLA-matched irradiated blood mononuclear cells in patients with renal cell carcinoma. J Clin Oncol, 2003, 21(20): 3785-3791.
100. Ertl HC, Xiang ZQ. Genetic immunization. Viral Immunol, 1996, 9(1): 1-9.
101. Takayama T, Sekine T, Makuuchi M, et al. Adoptive immunotherapy to lower postsurgical recurrence rates of hepatocellular carcinoma: a randomised trial. Lancet, 2000, 356(9232): 802-807.
102. Wagner U, Schlebusch H, Kohler S, et al. Immunological responses to the tumor-associated antigen CA125 in patients with advanced ovarian cancer induced by the murine monoclonal anti-idiotype vaccine ACA125. Hybridoma, 1997, 16(1): 33-40.
103. Talmor M, Mirza A, Turley S, et al. Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures. Eur J Immunol, 1998, 28(3): 811-817.
104. Winzler C, Rovere P, . Rescigno M, et al. Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures. J Exp Med, 1997, 185(2): 317-328.
105.陈向荣,石汉平.树突状细胞的分离纯化与鉴定.上海免疫学杂志,2001,21(2):122-125.
106. Brenan M, Puklavec M. The MRC OX-62 antigen: a useful marker in the purification of rat veiled cells with the biochemical properties of an integrin. J Exp Med, 1992, 175(6): 1457-1465.