冻融肿瘤疫苗的制备及对小鼠恶性黑色素瘤抑瘤作用的实验研究
|
摘要
目的：探讨冻融肿瘤疫苗对C57BL/6J小鼠黑色素瘤的抑制作用。
方法：(1)将B16Fo恶性黑色素瘤细胞皮下接种C57BL/6J小鼠建立肿瘤模型后,行氩氦刀冷冻消融肿瘤组织。(2)取冻融肿瘤组织联合弗氏不完全免疫佐剂制成肿瘤疫苗。(3)12只C57BL/6J小鼠随机分为实验组和对照组,每组6只。实验组小鼠注射肿瘤疫苗,每周1次,共4次,并于每次注射疫苗后眼眶采血,测量外周血CD4+、CD8+免疫指标变化；对照组小鼠注射佐剂和生理盐水。第四次疫苗注射完成后各组小鼠均皮下接种黑色素瘤细胞,观察各组小鼠的成瘤率、出瘤时间,肿瘤生长状态以及生存期。
结果：1.实验组及对照组小鼠全部成瘤,成瘤率为100%。实验组和对照组小鼠平均出瘤时间分别为11.35±1.6天和6.7±1.2天,两组比较差异有显著性(P＜0.01)；平均生存时间分别为36.0±2.8天和25.6±1.5天,两组比较差异有显著性(P＜0.01)。
2.实验组小鼠外周血T细胞亚群在免疫后呈动态变化,CD4+、CD4+／CD8+比值在免疫后逐渐增加,在第2周时与疫苗接种前相比差异有显著性(P＜0.01),在第4周时达到高峰；CDs+在免疫后开始下降,在第4周时与疫苗接种前相比差异有显著性(P＜0.01)。
结论：1.冻融肿瘤疫苗能有效延缓小鼠恶性黑色素瘤的生长,延长荷瘤小鼠的生存期。
2.冻融肿瘤疫苗能刺激机体细胞免疫,改善机体的免疫状况。
Objective:To investigate the inhibitory effect of tumor vaccines(the necrotic tumor tissues after cryosurgery) in mice with malignant melanoma carcinoma.
Methods:(1)The tumor model was established by subcutaneous implantation of B16Fo cells in C57BL/6J mice,then treated with cryosurgery.(2)Frozen tumor tissues combined with incomplete Freund adjuvant were made into tumor Vaccines.(3)Twelve C57BL/6J mice were randomly divided into 2 groups:experimental group and control group.Experimental mice group was injected with tumor Vaccines and blood was taken from the orbit after each vaccination,then the changes of CD4+、CD8+ were analyzed. Control mice group was injected with Adjuvant and NS.B16Fo cells was implanted subcutaneously into mice after 4th vaccination. The tumor growth and survival period were observed.
Results:1.The tumor formation rate of each group was 100%. The mean tumor growth time of experimental and control group were 11.3±1.6 and 6.7d±1.2 days,respectively(P<0.01),and the mean survival time of two group mice were 36.0±2.8 and 25.6d±1.5 days,respectively (P<0.01).
2.1n experimental group, T cell subsets in peripheral blood after immunization showed dynamic changes.CD4+、CD47/CD8+ gradually increased and CD8+ decreased after immunization,and reached a peak value in the 4th week(P<0.01).
Conclutions:1.Tumor vaccine can effectively delay the growth of malignant melanoma carcinoma and prolong survival time of immunized mices.
2.Tumor vaccine can stimulate body cell immunity and improve immune function.
方法：(1)将B16Fo恶性黑色素瘤细胞皮下接种C57BL/6J小鼠建立肿瘤模型后,行氩氦刀冷冻消融肿瘤组织。(2)取冻融肿瘤组织联合弗氏不完全免疫佐剂制成肿瘤疫苗。(3)12只C57BL/6J小鼠随机分为实验组和对照组,每组6只。实验组小鼠注射肿瘤疫苗,每周1次,共4次,并于每次注射疫苗后眼眶采血,测量外周血CD4+、CD8+免疫指标变化；对照组小鼠注射佐剂和生理盐水。第四次疫苗注射完成后各组小鼠均皮下接种黑色素瘤细胞,观察各组小鼠的成瘤率、出瘤时间,肿瘤生长状态以及生存期。
结果：1.实验组及对照组小鼠全部成瘤,成瘤率为100%。实验组和对照组小鼠平均出瘤时间分别为11.35±1.6天和6.7±1.2天,两组比较差异有显著性(P＜0.01)；平均生存时间分别为36.0±2.8天和25.6±1.5天,两组比较差异有显著性(P＜0.01)。
2.实验组小鼠外周血T细胞亚群在免疫后呈动态变化,CD4+、CD4+／CD8+比值在免疫后逐渐增加,在第2周时与疫苗接种前相比差异有显著性(P＜0.01),在第4周时达到高峰；CDs+在免疫后开始下降,在第4周时与疫苗接种前相比差异有显著性(P＜0.01)。
结论：1.冻融肿瘤疫苗能有效延缓小鼠恶性黑色素瘤的生长,延长荷瘤小鼠的生存期。
2.冻融肿瘤疫苗能刺激机体细胞免疫,改善机体的免疫状况。
Objective:To investigate the inhibitory effect of tumor vaccines(the necrotic tumor tissues after cryosurgery) in mice with malignant melanoma carcinoma.
Methods:(1)The tumor model was established by subcutaneous implantation of B16Fo cells in C57BL/6J mice,then treated with cryosurgery.(2)Frozen tumor tissues combined with incomplete Freund adjuvant were made into tumor Vaccines.(3)Twelve C57BL/6J mice were randomly divided into 2 groups:experimental group and control group.Experimental mice group was injected with tumor Vaccines and blood was taken from the orbit after each vaccination,then the changes of CD4+、CD8+ were analyzed. Control mice group was injected with Adjuvant and NS.B16Fo cells was implanted subcutaneously into mice after 4th vaccination. The tumor growth and survival period were observed.
Results:1.The tumor formation rate of each group was 100%. The mean tumor growth time of experimental and control group were 11.3±1.6 and 6.7d±1.2 days,respectively(P<0.01),and the mean survival time of two group mice were 36.0±2.8 and 25.6d±1.5 days,respectively (P<0.01).
2.1n experimental group, T cell subsets in peripheral blood after immunization showed dynamic changes.CD4+、CD47/CD8+ gradually increased and CD8+ decreased after immunization,and reached a peak value in the 4th week(P<0.01).
Conclutions:1.Tumor vaccine can effectively delay the growth of malignant melanoma carcinoma and prolong survival time of immunized mices.
2.Tumor vaccine can stimulate body cell immunity and improve immune function.
引文
[1]Dolan BP,Gibbs KD Jr, Ostrand-Rosenberg S. Tumor-specific CD4+T cells are activated by "cross-dressed" dendritic cells presenting peptide-MHC class II complexes acquired from cell based cancer Vaccines[J].J Immunol,2006,176(3):1447-55
[2]Armstrong TD,Pulaski BA,Ostrand-Rosemberg S, et al.Tumor antigen presentation:changing the rules[J].Cancer Immunaol Immunother,199 8,46(2):70-4
[3]Gabrilovich D,Pisarev V. Tumor escape from immune response:mechan-isms and targets of activity[J].Curr Drug Targets,2003,4(7):525-536
[4]Smyth MJ, Hayakawa y, Takeda K,et al.New aspects of natural-Killer-cell surveillance and therapy of cancer[J].Nat Rev Cancer,2002, 2(11):850-861
[5]张彩,田志刚.肿瘤免疫逃逸机制的研究进展.国外医学肿瘤学分册[J],2000,27(2)： 77-79
[6]Bullani RR,Wehrli P,Viard-Leveugle I, et al.Frequent downregulati-on of Fas(CD95) expression and function in melanoma[J].Melanoma Res,2002,12(3):263-70
[7]Quesnel B. Tumor dormancy and immunoescape[J].APMIS,2008,116(7-8): 685-94
[8]Teng MW, Swann JB,et al.Immune-mediated dormancy:an equilibrium with cancer[J].J Leukoc Biol,2008,84(4):988-93
[9]Tu SM, Lin SH, Logofitetis CJ.Stem-cell origin of metastasis and heterogeneity in solid tumors[J].Lancet Oncol,2002,3(8):508-13
[10]Lawson JC,Blatch GL,Edkins AL. Cancer stem cells in breast cancer and metastasis[J].Breast cancer Res Treat,2009,118(2):241-54
[11]Dermime S,Armstrong A, et al. Cancer vaccines and immunotherapy.[J] Br Med Bull,2002,62:149-162
[12]杜娟,钱晓萍,刘宝瑞.肿瘤疫苗的研究研究进展[J].现代肿瘤学,2009.17(1)：161-162
[13]Banat GA,Christ 0,et al.Tumor-induced suppression of immune res- ponse and its correction[J].Cancer Immunol Immunother,2001,49 (11):573-86
[14]Cavallo F, Forni G. Recent advances in cancer immunotherapy with an emphasis on vaccines[J].Expert Rev Vaccines,2009,8(1):25-8
[15]盛秋菊,王洪涛,宋永喜.治疗性肿瘤疫苗的研究进展[J].现代医药卫生,2007,23(15)：2284-2287
[16]Hockertz S. Present and future of cancer vaccines[J].Toxicology,2 005,214(1-2):151-161
[17]Goldman B. Cancer vaccines:finding the best way to train the imm-ne system[J].J Nati cancer Inst,2002,94(20):1523-6
[18]Tabi Z,Man S. Challenges for cancer vaccine development[J].Adv Drug Deli.v Rev,2006,58(8):902-15
[19]Uyl-de Groot CA,Vermorken JB,Hanna MG Jr,et al.Immunotherapy with autologous tumor cell-BCG vaccine in patients with colon cancer:a prospective study of medical and economic benefits [J].Vaccine,2005,23(17-18):2379-2383
[20]张银旭,张俊华,等.自体肿瘤细胞疫苗在结肠癌术后的主动免疫治疗研究[J].中国医师进修杂志,2007,30(3)：17-19
[21]Joeham D,Richter A,Hoffmann L,etal.Adjuvant autologous renal tumourcell vaccine and risk of tarnour progression in patients witll renal-cell carcinoma after radical nephreetomy:phase nl, randomised controlled trial[J].Lancet,2004,363(9409):594-599
[22]Gage AA.History of cryesurgery[J].Semin Surg Oncol,1998,14(2):99-109
[23]汤钊猷.现代肿瘤学[M].上海：上海医科大学出版社,1993.1081-1083
[24]van der Bruggen P, Chomez P, et al.A gene encoding an antigen rec-ognized by cytolytic T lymphoeytes on a human melanoma[J].J immunol,2007,178(5):2617-21
[25]罗锋,魏于全.异种黑色素细胞疫苗诱导小鼠抗恶性黑色素瘤免疫效应[J].中华肿瘤学杂志,2001,23(2)：118-122
[26]马飞,孙文欣,等.SLC基因修饰对肿瘤细胞疫苗免疫效果的初步研究[J].中华肿瘤预防杂志,2008,15(5)：321-325
[27]Berd D.Contribution of dead cells to the immunogenicity of an autologous,hapten-modified melanoma vaccine[J].Vaccine,2003, 21(7-8):795-797
[28]Vaishampayan U,Abrams J,Darrah D, et al.Acitve immunotherapy of metastatic melanoma with allogeneic melanoma lysates and interf-eron alpha[J].Clin Cancer Res,2002,8(12):3696-701
[29]陈慰峰.医学免疫学[M].北京：人民卫生出版社,2001.92-93
[30]Kim TY, Myoung HJ, et al.Both E7 and CpG-Oligodeoxy-nucleotide are required for protective immunity against challenge with human papillomavirus 16(E6/E7)immortalized tumor cells:involvement of CD4+Tand CD8+T cells in protection[J].Cancer Res,2002,62(24):72 34-7240
[31]Kennedy RC, Shearer MH, Watts AM, et al.CD4+T lymphocytes play a role in antibody production and tumor immunity against simian virus 40 large tumor antigen[J].Cancer Res,2003,63(5):1040-1045
[32]李挺,刘剑勇,等.肝癌疫苗激活的肿瘤侵润性淋巴细胞治疗原发性肝癌的研究[J].肝脏普通外科杂志,2002,17(4)：202-203
[33]王仙,李永祥,等.NDV/β-榄香烯自体瘤苗对消化道肿瘤患者T细胞不同亚群平衡的影响[J].Modern oncology,2008,16(4):606-609
[1]Slominski A, Wortsman J, Carlson AJ, et al.Malignant melanoma[J]. ArchPathol Lab Med,2001,125(10):295-306
[2]陈德谊.黑色素瘤胃肠道转移的手术切除[J].国外医学外科学分册,1997,24(5)：101
[3]Ozyuncu N,Sahin M,Altin T, et al.Cardiac metastasis of malignant melanoma:a rare cause of complete atrioventricular block[J].Eur-opace,2006,8(7):545-548
[4]Gorg C,Ramaswamy A,Barth PJ,et al.Splenic metastases in a large unselected autopsy series[J].Pathol Res Pract,2006,202(5):351-356
[5]Schnaeker EM,Ossig R,Ludwig T,et al.Microtubuel-dependent matrix metalloproteinase-2/matrix metalloproteinase-9 exocytosis:prere-quisite in human melanoma cell invasion[J].Cancer Res,2004,64(24) :8924-8931
[6]Mohammed MQ,Abraha HD,Sherwood RA,et al.Serum S100 beta protein as a marker of disease activity in patients with malignant mela-noma[J].Med Oncol,2001,18(2):109
[7]Daycl Jr Mihm MC Jr Sober AJ,et al.Narrower margins for clinical stage I malignant melanoma[J].N EngJ Med,1982,306(2):497
[8]Thompson JA. The revised American Joint Committee on Cancer stag-ing system for melanoma[J].Semin Oncol,2002,29(4):361
[9]储大同.当代肿瘤内科治疗方案评价[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.173-176
[10]Morton DL, Foshag LJ, et al.Active specific immunotherapy in mali-gnant melanoma[J].Semin Surg Oncol,1989,5(6):420-5
[11]Morton DL. Immune response to postsurgical adjuvant active immun-otherapy with Canvaxin polyvalent cancer vaccine:correlations with clinical course of patients with metastatic melanoma[J].Dev Biol(Basel),2004,116:209-217
[12]Menon C, Iyer M, Prabakaran I,et al.TNF-alpha dowuregulates vascu-lar endothetial Flk-1 expression in human melanoma xenograft mo-del[J].Am J Physiol Heart Cire Physiol,2003,284(1):H317-29
[13]Fujita M,Mason RJ, et al.Pulmonary hypertension in TNF-alpha-Overexpressing mice is associated with decreased VEGF gene Expression[J].J Appl Physiol,2002,93(6):2162-70
[14]Rosenberg SA, Lotze MT, et al.Prospective randomized trial of high-dose interleukin-2 along or in conjunction with lymphokine-actived Killer cells for the treatment of patients with advanced cancer[J].J Natl cancer Inst,1993,85(8):622-32
[15]Choi KS, Song EK,Yim CY.Cytokines secreted by IL-2-activated lym-Phocytes induce endogenous nitric oxide synthesis and apoptosis in macrophages[J].J Leukoc Biol,2008,83(6):1440-50
[16]Oble DA, Loewe R, et al.Focus on TILs:prognostic significance of tumor infiltrating lymphocytes in human melanoma[J].Cancer Immun,2009,9:3
[17]唐佩福、曲国番等,恶性黑色素瘤TIL细胞免疫过继疗法途径的探讨[J].黑龙江医学,1999,3(177)：3
[18]Banchereau J, Briere F, Caux C, et al.Immunobiology of dendritic cells[J].Annu Rev Immunol,2000,18:767-811
[19]Morton DL. Immune response to postsurgical adjuvant immunotherapy With Canvaxin polyvalent cancer vaccine:correlations with clini-cal course of patients with metastatic melanoma[J].Dev Biol,2004, 116:209-17
[20]Trefzer U,Herberth G,Wohlan K,et al.Tumour-dendritic hybrid cell Vaccination for the treatment of patients with malignant melano-a:immunological effects and clinical results[J].Vaccine,2005, 23(17-18):2367-2373
[21]Bahri R, Naji A, et al.Dendritic cells secrete the immunosuppress-Ive HLA-G molecule upon CTLA4-Ig treatment:implication in human Renal transplant acceptance[J].J Immunol,2009,183(11):7054-62
[22]Phan GQ,Touloukian CE,Yang JC,el al..Immunization of patients With metastatic melanoma using both class I-and class II-restri-cted peptides from melanoma-associated antigens[J].J Immunother, 2003,26(4):349-356
[23]查园园,林晨等.基因枪介导的neu基因DNA疫苗抗肿瘤作用的实验研究[J].中华肿瘤杂志,2003,25(4),320-324
[24]Phan GQ,Touloukian CE,Yang JC,el al.Immunization of patients with metastatic melanoma using both class I-and class Ⅱ-restr-icted peptides from melanoma-associated antigens[J].J Immunother 2003,26(4):349-356
[25]Cytron S, Greene D, et al.Cryoablation of the prostate:technical recommendations[J].Prostate cancer Prostatic Dis,2009;12(4):339-46
[26]Yantorno C,Soanes WA,et al.Studies in cryo-immunology.I.The pro-Duction of antibodies to urogenital tissue in consequence of Freezing treatment[J].Immunology,1967,12(4):395-410
[27]Shulman S, Brandt EJ, Yantorno C.Studies in cryo-immunology.Ⅱ. Ti-Ssue and species specificity of the autoantibody response and Comparison with iso-immunization[J].Immunology,1968,14(2):149-15 8
[28]Osada S,Imai H,Tomita H,et al.Serum cytokine levels in response to hepatic cryoablation[J].J Surg Oncol,2007,95(6):491-498
[29]Si T, Guo z,Hao x.Immunologic response to primary cryoablation of High-risk prostate cancer[J].Cryobiology,2008,57(1):66-71
[30]Soanes WA.Gonder MJ.Ablin RJ.A possible immuno-cryothermic resp-onse in prostatic cancer[J].Clin Radiol,1970,21(3):253-255
[31]Abhn RJ,Soanes WA.Gonder MJ.Elution of in vlvo bound antiprosta-Tic epithetlial antibodies following multiple cryotherapy of carcinoma of prostate[J].Urology,1973,2(3):276-279
[32]周霖,王福生.氩氦刀冷冻治疗的抗肿瘤免疫效应研究进展[J].医学综述,2008,14(17)：2631-2633
[33]张克勤,张积仁.氩氦刀冷冻、射频消融和微波凝固对兔VX2肝癌消融作用的对比[J].广东医学,2008,29(2)：212-216
[34]Bagley DH,Farci RP, et al.Lymphocyte mediated cytotoxity after cryosurgery of a murine sarcoma[J].J Surg Res,1974,17:404-406
[35]Rauch D, Gross S, et al.T-cell activation promotes tumorigenesis In Inflammation-associated cancer[J].Retrovirology,2009,17(6):11 6
[36]杨宜娥,付听,吕莎.CD4+CD25+调节性T细胞在恶性肿瘤患者中的应用价值[J].医学检验与临床,2007,18：15-21
[37]钱建新,徐映东.淋巴细胞亚群与免疫的初探[J].免疫学杂志,1992,8：251
[38]李德忠,宋华志.氩氦刀冷冻兔肝后局部组织变化及超微病理研究[J].华南国防医学杂志,2006,20(3)：4-6
[39]Imai H, Saio M, Nonaka K, et al.Depletion of D4+CD25+ regulation T cells enhances interleukin-2-induced antitumor immunity in a mouse model of colon adenocarcinoma[J].Cancer Sci,2007,98(3):416-423
[40]Nair S, Boczkowski D, Fassnacht M, et al.Vaccination against the forkhead family transcription factor Foxp3 enhances tumor immun-ity[J].Cancer Res,2007,67(1):371-380
、[41]杨宝亮,张艳桥.氩氦刀治疗恶性肿瘤对细胞免疫功能影响[J].牡丹江医学报,2008,29：47-49
[42]Aarntzen EH,Figdor CG,Adema GJ,et aL.Dendritic cell vaccination and immune monitoring[J].Cancer Immunol lmmunother,2008,57(10): 1559-1568
[43]Tuettenberg A, Schmitt E, et al.Dendritic cell-based immunotherapy of Malignant:success and limitations[J].J Dtsch Dermatol Ges,200 7,5(3):190-6
[44]Den Brok MH,Sutmuller RP,NierkensS,et al.Synergy between in situ cryoablation and TLR9 stimulation results in a highly effective in vivo dendritic cell vaccine[J].Cancer Res,2006,66(14):7285-7292
[45]Den Bmk MH,Sutmuller RP,Nierkens S,et aL.Efficient loading of dendritic cells following cryo and radiofrequency ablation in combination with immune modulation induces anti-tumour immunity [J].Br J Cancer,2006,95(7):896-905
[46]韦长元,刘剑仑,张传珉,等.肝癌病人化疗栓塞后联合应用s-TIL细胞后T淋巴细胞亚群的变化[J].肿瘤研究与临床,2000,12(2)：681
[47]刘全则,刘作斌.低温学[M].北京：人民卫生出版社,1992.311-317
[48]孙燕,哈献文.关键治疗临床肿瘤[M].南宁：广西科学技术出版社,1999.308-309
[49]Hausen,Gruis NA,Weaver-Feldhaus J,et al.A cell cycleregulator potentially involveding enesisofmany tuner types [J].Science,1997 ,88:110
[50]蒙志斌.冷冻和激光为主综合治疗恶性黑色素瘤20例临床分析[J].广西医 学,1996,18(3)：262
[51]崔盘根,节译.恶性黑色素瘤的免疫治疗[J].国外医学皮肤病学分册,1997,19(2)：102
[52]Rosenberg SA,Spiess P,et al.A new approach to the adoptive immu-notherapy of cancer with tumor infiltrating lymphocytes[J]. Science,1986,233(4707):1318-1321
[53]de Jong RA, Leffers N, et al.Presence of tumor-infiltrating lymph-ocytes is an independen prognostic factor in type Ⅰ and Ⅱ endometrial cancer[J].Gynecol Oncol,2009,114(1):105-10
[54]蒙志斌,欧阳欣,欧盛秋,等.自体冷冻免疫瘤苗澈活肿瘤浸润淋巴细胞治疗恶性黑色索瘤的疗效观察[J].广西医学,2006,28(11)：1689-91
[55]Topalian SL, Muul LM, Solomon D, et al.Expansion of human tumor in-filtrating lymphocytes for use in immunotherapy trial[J].J Immunol Methods,1987,102(1):127-41
[56]蒙志斌,欧盛秋,等.晚期黑色素瘤免疫综合治疗的临床分析[J].中国癌症防治杂志,2009,9(1)：245-248
[57]Gage AA, Baust J. Mechanisms of tissue injury in cryosuegery[J]. Cryobiology,1998,37(3):171-186
[58]骆益宇、钱建新,等.氩氦刀治疗黑色素瘤对小鼠T细胞免疫的影响[J].华南国防医学杂志,2008,22(1)：7-8
[2]Armstrong TD,Pulaski BA,Ostrand-Rosemberg S, et al.Tumor antigen presentation:changing the rules[J].Cancer Immunaol Immunother,199 8,46(2):70-4
[3]Gabrilovich D,Pisarev V. Tumor escape from immune response:mechan-isms and targets of activity[J].Curr Drug Targets,2003,4(7):525-536
[4]Smyth MJ, Hayakawa y, Takeda K,et al.New aspects of natural-Killer-cell surveillance and therapy of cancer[J].Nat Rev Cancer,2002, 2(11):850-861
[5]张彩,田志刚.肿瘤免疫逃逸机制的研究进展.国外医学肿瘤学分册[J],2000,27(2)： 77-79
[6]Bullani RR,Wehrli P,Viard-Leveugle I, et al.Frequent downregulati-on of Fas(CD95) expression and function in melanoma[J].Melanoma Res,2002,12(3):263-70
[7]Quesnel B. Tumor dormancy and immunoescape[J].APMIS,2008,116(7-8): 685-94
[8]Teng MW, Swann JB,et al.Immune-mediated dormancy:an equilibrium with cancer[J].J Leukoc Biol,2008,84(4):988-93
[9]Tu SM, Lin SH, Logofitetis CJ.Stem-cell origin of metastasis and heterogeneity in solid tumors[J].Lancet Oncol,2002,3(8):508-13
[10]Lawson JC,Blatch GL,Edkins AL. Cancer stem cells in breast cancer and metastasis[J].Breast cancer Res Treat,2009,118(2):241-54
[11]Dermime S,Armstrong A, et al. Cancer vaccines and immunotherapy.[J] Br Med Bull,2002,62:149-162
[12]杜娟,钱晓萍,刘宝瑞.肿瘤疫苗的研究研究进展[J].现代肿瘤学,2009.17(1)：161-162
[13]Banat GA,Christ 0,et al.Tumor-induced suppression of immune res- ponse and its correction[J].Cancer Immunol Immunother,2001,49 (11):573-86
[14]Cavallo F, Forni G. Recent advances in cancer immunotherapy with an emphasis on vaccines[J].Expert Rev Vaccines,2009,8(1):25-8
[15]盛秋菊,王洪涛,宋永喜.治疗性肿瘤疫苗的研究进展[J].现代医药卫生,2007,23(15)：2284-2287
[16]Hockertz S. Present and future of cancer vaccines[J].Toxicology,2 005,214(1-2):151-161
[17]Goldman B. Cancer vaccines:finding the best way to train the imm-ne system[J].J Nati cancer Inst,2002,94(20):1523-6
[18]Tabi Z,Man S. Challenges for cancer vaccine development[J].Adv Drug Deli.v Rev,2006,58(8):902-15
[19]Uyl-de Groot CA,Vermorken JB,Hanna MG Jr,et al.Immunotherapy with autologous tumor cell-BCG vaccine in patients with colon cancer:a prospective study of medical and economic benefits [J].Vaccine,2005,23(17-18):2379-2383
[20]张银旭,张俊华,等.自体肿瘤细胞疫苗在结肠癌术后的主动免疫治疗研究[J].中国医师进修杂志,2007,30(3)：17-19
[21]Joeham D,Richter A,Hoffmann L,etal.Adjuvant autologous renal tumourcell vaccine and risk of tarnour progression in patients witll renal-cell carcinoma after radical nephreetomy:phase nl, randomised controlled trial[J].Lancet,2004,363(9409):594-599
[22]Gage AA.History of cryesurgery[J].Semin Surg Oncol,1998,14(2):99-109
[23]汤钊猷.现代肿瘤学[M].上海：上海医科大学出版社,1993.1081-1083
[24]van der Bruggen P, Chomez P, et al.A gene encoding an antigen rec-ognized by cytolytic T lymphoeytes on a human melanoma[J].J immunol,2007,178(5):2617-21
[25]罗锋,魏于全.异种黑色素细胞疫苗诱导小鼠抗恶性黑色素瘤免疫效应[J].中华肿瘤学杂志,2001,23(2)：118-122
[26]马飞,孙文欣,等.SLC基因修饰对肿瘤细胞疫苗免疫效果的初步研究[J].中华肿瘤预防杂志,2008,15(5)：321-325
[27]Berd D.Contribution of dead cells to the immunogenicity of an autologous,hapten-modified melanoma vaccine[J].Vaccine,2003, 21(7-8):795-797
[28]Vaishampayan U,Abrams J,Darrah D, et al.Acitve immunotherapy of metastatic melanoma with allogeneic melanoma lysates and interf-eron alpha[J].Clin Cancer Res,2002,8(12):3696-701
[29]陈慰峰.医学免疫学[M].北京：人民卫生出版社,2001.92-93
[30]Kim TY, Myoung HJ, et al.Both E7 and CpG-Oligodeoxy-nucleotide are required for protective immunity against challenge with human papillomavirus 16(E6/E7)immortalized tumor cells:involvement of CD4+Tand CD8+T cells in protection[J].Cancer Res,2002,62(24):72 34-7240
[31]Kennedy RC, Shearer MH, Watts AM, et al.CD4+T lymphocytes play a role in antibody production and tumor immunity against simian virus 40 large tumor antigen[J].Cancer Res,2003,63(5):1040-1045
[32]李挺,刘剑勇,等.肝癌疫苗激活的肿瘤侵润性淋巴细胞治疗原发性肝癌的研究[J].肝脏普通外科杂志,2002,17(4)：202-203
[33]王仙,李永祥,等.NDV/β-榄香烯自体瘤苗对消化道肿瘤患者T细胞不同亚群平衡的影响[J].Modern oncology,2008,16(4):606-609
[1]Slominski A, Wortsman J, Carlson AJ, et al.Malignant melanoma[J]. ArchPathol Lab Med,2001,125(10):295-306
[2]陈德谊.黑色素瘤胃肠道转移的手术切除[J].国外医学外科学分册,1997,24(5)：101
[3]Ozyuncu N,Sahin M,Altin T, et al.Cardiac metastasis of malignant melanoma:a rare cause of complete atrioventricular block[J].Eur-opace,2006,8(7):545-548
[4]Gorg C,Ramaswamy A,Barth PJ,et al.Splenic metastases in a large unselected autopsy series[J].Pathol Res Pract,2006,202(5):351-356
[5]Schnaeker EM,Ossig R,Ludwig T,et al.Microtubuel-dependent matrix metalloproteinase-2/matrix metalloproteinase-9 exocytosis:prere-quisite in human melanoma cell invasion[J].Cancer Res,2004,64(24) :8924-8931
[6]Mohammed MQ,Abraha HD,Sherwood RA,et al.Serum S100 beta protein as a marker of disease activity in patients with malignant mela-noma[J].Med Oncol,2001,18(2):109
[7]Daycl Jr Mihm MC Jr Sober AJ,et al.Narrower margins for clinical stage I malignant melanoma[J].N EngJ Med,1982,306(2):497
[8]Thompson JA. The revised American Joint Committee on Cancer stag-ing system for melanoma[J].Semin Oncol,2002,29(4):361
[9]储大同.当代肿瘤内科治疗方案评价[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.173-176
[10]Morton DL, Foshag LJ, et al.Active specific immunotherapy in mali-gnant melanoma[J].Semin Surg Oncol,1989,5(6):420-5
[11]Morton DL. Immune response to postsurgical adjuvant active immun-otherapy with Canvaxin polyvalent cancer vaccine:correlations with clinical course of patients with metastatic melanoma[J].Dev Biol(Basel),2004,116:209-217
[12]Menon C, Iyer M, Prabakaran I,et al.TNF-alpha dowuregulates vascu-lar endothetial Flk-1 expression in human melanoma xenograft mo-del[J].Am J Physiol Heart Cire Physiol,2003,284(1):H317-29
[13]Fujita M,Mason RJ, et al.Pulmonary hypertension in TNF-alpha-Overexpressing mice is associated with decreased VEGF gene Expression[J].J Appl Physiol,2002,93(6):2162-70
[14]Rosenberg SA, Lotze MT, et al.Prospective randomized trial of high-dose interleukin-2 along or in conjunction with lymphokine-actived Killer cells for the treatment of patients with advanced cancer[J].J Natl cancer Inst,1993,85(8):622-32
[15]Choi KS, Song EK,Yim CY.Cytokines secreted by IL-2-activated lym-Phocytes induce endogenous nitric oxide synthesis and apoptosis in macrophages[J].J Leukoc Biol,2008,83(6):1440-50
[16]Oble DA, Loewe R, et al.Focus on TILs:prognostic significance of tumor infiltrating lymphocytes in human melanoma[J].Cancer Immun,2009,9:3
[17]唐佩福、曲国番等,恶性黑色素瘤TIL细胞免疫过继疗法途径的探讨[J].黑龙江医学,1999,3(177)：3
[18]Banchereau J, Briere F, Caux C, et al.Immunobiology of dendritic cells[J].Annu Rev Immunol,2000,18:767-811
[19]Morton DL. Immune response to postsurgical adjuvant immunotherapy With Canvaxin polyvalent cancer vaccine:correlations with clini-cal course of patients with metastatic melanoma[J].Dev Biol,2004, 116:209-17
[20]Trefzer U,Herberth G,Wohlan K,et al.Tumour-dendritic hybrid cell Vaccination for the treatment of patients with malignant melano-a:immunological effects and clinical results[J].Vaccine,2005, 23(17-18):2367-2373
[21]Bahri R, Naji A, et al.Dendritic cells secrete the immunosuppress-Ive HLA-G molecule upon CTLA4-Ig treatment:implication in human Renal transplant acceptance[J].J Immunol,2009,183(11):7054-62
[22]Phan GQ,Touloukian CE,Yang JC,el al..Immunization of patients With metastatic melanoma using both class I-and class II-restri-cted peptides from melanoma-associated antigens[J].J Immunother, 2003,26(4):349-356
[23]查园园,林晨等.基因枪介导的neu基因DNA疫苗抗肿瘤作用的实验研究[J].中华肿瘤杂志,2003,25(4),320-324
[24]Phan GQ,Touloukian CE,Yang JC,el al.Immunization of patients with metastatic melanoma using both class I-and class Ⅱ-restr-icted peptides from melanoma-associated antigens[J].J Immunother 2003,26(4):349-356
[25]Cytron S, Greene D, et al.Cryoablation of the prostate:technical recommendations[J].Prostate cancer Prostatic Dis,2009;12(4):339-46
[26]Yantorno C,Soanes WA,et al.Studies in cryo-immunology.I.The pro-Duction of antibodies to urogenital tissue in consequence of Freezing treatment[J].Immunology,1967,12(4):395-410
[27]Shulman S, Brandt EJ, Yantorno C.Studies in cryo-immunology.Ⅱ. Ti-Ssue and species specificity of the autoantibody response and Comparison with iso-immunization[J].Immunology,1968,14(2):149-15 8
[28]Osada S,Imai H,Tomita H,et al.Serum cytokine levels in response to hepatic cryoablation[J].J Surg Oncol,2007,95(6):491-498
[29]Si T, Guo z,Hao x.Immunologic response to primary cryoablation of High-risk prostate cancer[J].Cryobiology,2008,57(1):66-71
[30]Soanes WA.Gonder MJ.Ablin RJ.A possible immuno-cryothermic resp-onse in prostatic cancer[J].Clin Radiol,1970,21(3):253-255
[31]Abhn RJ,Soanes WA.Gonder MJ.Elution of in vlvo bound antiprosta-Tic epithetlial antibodies following multiple cryotherapy of carcinoma of prostate[J].Urology,1973,2(3):276-279
[32]周霖,王福生.氩氦刀冷冻治疗的抗肿瘤免疫效应研究进展[J].医学综述,2008,14(17)：2631-2633
[33]张克勤,张积仁.氩氦刀冷冻、射频消融和微波凝固对兔VX2肝癌消融作用的对比[J].广东医学,2008,29(2)：212-216
[34]Bagley DH,Farci RP, et al.Lymphocyte mediated cytotoxity after cryosurgery of a murine sarcoma[J].J Surg Res,1974,17:404-406
[35]Rauch D, Gross S, et al.T-cell activation promotes tumorigenesis In Inflammation-associated cancer[J].Retrovirology,2009,17(6):11 6
[36]杨宜娥,付听,吕莎.CD4+CD25+调节性T细胞在恶性肿瘤患者中的应用价值[J].医学检验与临床,2007,18：15-21
[37]钱建新,徐映东.淋巴细胞亚群与免疫的初探[J].免疫学杂志,1992,8：251
[38]李德忠,宋华志.氩氦刀冷冻兔肝后局部组织变化及超微病理研究[J].华南国防医学杂志,2006,20(3)：4-6
[39]Imai H, Saio M, Nonaka K, et al.Depletion of D4+CD25+ regulation T cells enhances interleukin-2-induced antitumor immunity in a mouse model of colon adenocarcinoma[J].Cancer Sci,2007,98(3):416-423
[40]Nair S, Boczkowski D, Fassnacht M, et al.Vaccination against the forkhead family transcription factor Foxp3 enhances tumor immun-ity[J].Cancer Res,2007,67(1):371-380
、[41]杨宝亮,张艳桥.氩氦刀治疗恶性肿瘤对细胞免疫功能影响[J].牡丹江医学报,2008,29：47-49
[42]Aarntzen EH,Figdor CG,Adema GJ,et aL.Dendritic cell vaccination and immune monitoring[J].Cancer Immunol lmmunother,2008,57(10): 1559-1568
[43]Tuettenberg A, Schmitt E, et al.Dendritic cell-based immunotherapy of Malignant:success and limitations[J].J Dtsch Dermatol Ges,200 7,5(3):190-6
[44]Den Brok MH,Sutmuller RP,NierkensS,et al.Synergy between in situ cryoablation and TLR9 stimulation results in a highly effective in vivo dendritic cell vaccine[J].Cancer Res,2006,66(14):7285-7292
[45]Den Bmk MH,Sutmuller RP,Nierkens S,et aL.Efficient loading of dendritic cells following cryo and radiofrequency ablation in combination with immune modulation induces anti-tumour immunity [J].Br J Cancer,2006,95(7):896-905
[46]韦长元,刘剑仑,张传珉,等.肝癌病人化疗栓塞后联合应用s-TIL细胞后T淋巴细胞亚群的变化[J].肿瘤研究与临床,2000,12(2)：681
[47]刘全则,刘作斌.低温学[M].北京：人民卫生出版社,1992.311-317
[48]孙燕,哈献文.关键治疗临床肿瘤[M].南宁：广西科学技术出版社,1999.308-309
[49]Hausen,Gruis NA,Weaver-Feldhaus J,et al.A cell cycleregulator potentially involveding enesisofmany tuner types [J].Science,1997 ,88:110
[50]蒙志斌.冷冻和激光为主综合治疗恶性黑色素瘤20例临床分析[J].广西医 学,1996,18(3)：262
[51]崔盘根,节译.恶性黑色素瘤的免疫治疗[J].国外医学皮肤病学分册,1997,19(2)：102
[52]Rosenberg SA,Spiess P,et al.A new approach to the adoptive immu-notherapy of cancer with tumor infiltrating lymphocytes[J]. Science,1986,233(4707):1318-1321
[53]de Jong RA, Leffers N, et al.Presence of tumor-infiltrating lymph-ocytes is an independen prognostic factor in type Ⅰ and Ⅱ endometrial cancer[J].Gynecol Oncol,2009,114(1):105-10
[54]蒙志斌,欧阳欣,欧盛秋,等.自体冷冻免疫瘤苗澈活肿瘤浸润淋巴细胞治疗恶性黑色索瘤的疗效观察[J].广西医学,2006,28(11)：1689-91
[55]Topalian SL, Muul LM, Solomon D, et al.Expansion of human tumor in-filtrating lymphocytes for use in immunotherapy trial[J].J Immunol Methods,1987,102(1):127-41
[56]蒙志斌,欧盛秋,等.晚期黑色素瘤免疫综合治疗的临床分析[J].中国癌症防治杂志,2009,9(1)：245-248
[57]Gage AA, Baust J. Mechanisms of tissue injury in cryosuegery[J]. Cryobiology,1998,37(3):171-186
[58]骆益宇、钱建新,等.氩氦刀治疗黑色素瘤对小鼠T细胞免疫的影响[J].华南国防医学杂志,2008,22(1)：7-8