肿瘤抗原MUC4 HLA-A~*0201限制性CTL表位的体外免疫效应研究
摘要
目的:探讨肿瘤抗原MUC4 HLA-A*0201限制性CTL表位肽负载树突状细胞诱导细胞毒性T淋巴细胞(CTL)的体外免疫效应。
方法:从外周血单个核细胞(PBMC)中诱导树突状细胞(DC)。用之前表位预测法预测并合成的五种肽分子分别脉冲成熟的DC,并刺激HLA-A*0201~+健康人自体CD8~+T细胞成为CTL。周酶联免疫斑点法(ELISPOT),检测CTL IFN-γ的分泌。同时用乳酸脱氢酶(LDH)释放法检测其对靶细胞的杀伤作用。
结果:人PBMC可在多种细胞因子的作用下被诱导成为DC。肽P01204诱导的CTL,在不同的效靶比可特异性杀伤P01204脉冲的T2细胞(5:1时为9.03%±0.24%,10:1时为19.62%±0.44%,20:1时为27.93%±2.22%)和MUC4~+、HLA-A2~+的HCT-116细胞(5:1时为7.26%±0.18%,10:1时为16.83%±0.40%,20:1时为25.23%±1.35%),均高于阴性对照组(P＜0.05),能够产生IFN-γ的细胞数(130.33±6.58)明显高于阴性对照组(P＜0.05)。
结论:肿瘤相关抗原MUC4的HLA-A*0201限制性CTL表位P01204,能诱导人外周血单核细胞的CTL反应,该活化的CTL能分泌免疫活性物质诱导特异性靶细胞的凋亡。
Objective: To explore the in vitro immune response to tumor associated antigen MUC4-derived cytotoxic T lymphocyte (CTL) epitope-pulsed dendritic cell (DC).
Methods: DCs was induced from the HLA-A*0201-positive healthy individuals' peripheral blood mononuclear cells (PBMC). DCs' phenotype was identified by flow cytometry. CD8~+ T cells were isolated and purified through MACS (magnetic activated cell sorting). Mature DCs were pulsed with each of the five synthesized peptides which were selected as possible CTL epitopes by software analysis previously. Autologous CD8~+ T cells from the HLA-A* 0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase (LDH) release assay and IFN-γreleased by enzyme-linked immuno spot assay (ELISPOT).
Results: Mature DCs could be induced from PBMCs through several cytokines' action. CTL induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204 (9.03%±0.24%,19.62%±0.44%, 27.93%±2.22% at ratios of 5:1,10:1,20:1) and HCT-116 cells (MUC4~+, HLA-A2~+) (7.26%±0.18%,16.83%±0.40%,25.23%±1.35% at ratios of 5:1,10:1,20:1). The number of CTL induced by peptide P01204 which could secret IFN-γ(130.33±6.58) was obviously higher than the negative group.
Conclusion: Tumor associated antigen MUC4-derived HLA-A*0201-restrictive cytotoxic T lymphocyte (CTL) epitope P01204 could induce PBMCs' CTL reaction. The CTL could secret immunologic active material to induce the specific target cells' lysis.
方法:从外周血单个核细胞(PBMC)中诱导树突状细胞(DC)。用之前表位预测法预测并合成的五种肽分子分别脉冲成熟的DC,并刺激HLA-A*0201~+健康人自体CD8~+T细胞成为CTL。周酶联免疫斑点法(ELISPOT),检测CTL IFN-γ的分泌。同时用乳酸脱氢酶(LDH)释放法检测其对靶细胞的杀伤作用。
结果:人PBMC可在多种细胞因子的作用下被诱导成为DC。肽P01204诱导的CTL,在不同的效靶比可特异性杀伤P01204脉冲的T2细胞(5:1时为9.03%±0.24%,10:1时为19.62%±0.44%,20:1时为27.93%±2.22%)和MUC4~+、HLA-A2~+的HCT-116细胞(5:1时为7.26%±0.18%,10:1时为16.83%±0.40%,20:1时为25.23%±1.35%),均高于阴性对照组(P＜0.05),能够产生IFN-γ的细胞数(130.33±6.58)明显高于阴性对照组(P＜0.05)。
结论:肿瘤相关抗原MUC4的HLA-A*0201限制性CTL表位P01204,能诱导人外周血单核细胞的CTL反应,该活化的CTL能分泌免疫活性物质诱导特异性靶细胞的凋亡。
Objective: To explore the in vitro immune response to tumor associated antigen MUC4-derived cytotoxic T lymphocyte (CTL) epitope-pulsed dendritic cell (DC).
Methods: DCs was induced from the HLA-A*0201-positive healthy individuals' peripheral blood mononuclear cells (PBMC). DCs' phenotype was identified by flow cytometry. CD8~+ T cells were isolated and purified through MACS (magnetic activated cell sorting). Mature DCs were pulsed with each of the five synthesized peptides which were selected as possible CTL epitopes by software analysis previously. Autologous CD8~+ T cells from the HLA-A* 0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase (LDH) release assay and IFN-γreleased by enzyme-linked immuno spot assay (ELISPOT).
Results: Mature DCs could be induced from PBMCs through several cytokines' action. CTL induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204 (9.03%±0.24%,19.62%±0.44%, 27.93%±2.22% at ratios of 5:1,10:1,20:1) and HCT-116 cells (MUC4~+, HLA-A2~+) (7.26%±0.18%,16.83%±0.40%,25.23%±1.35% at ratios of 5:1,10:1,20:1). The number of CTL induced by peptide P01204 which could secret IFN-γ(130.33±6.58) was obviously higher than the negative group.
Conclusion: Tumor associated antigen MUC4-derived HLA-A*0201-restrictive cytotoxic T lymphocyte (CTL) epitope P01204 could induce PBMCs' CTL reaction. The CTL could secret immunologic active material to induce the specific target cells' lysis.
引文
1 Byrd J C,Bresalier R S.Mucins and mucin binding proteins in colorectal[J].Cancer Metastasis Rev,2004,23(1-2):77-99.
2 汪荣泉,房殿春,刘为纹,等.用BC2单抗检测正常腺上皮组织中表达MUC1基因的表达[J].第三军医大学学报,1999,21(6):413-415.
3 汪荣泉,房殿春,罗元辉,等.胃癌及癌前病变组织中MUC5AC基因的异常表达及意义[J].癌症,2000,19(2):121-123.
4 Aksoy N,Akinci O F.Mucin macromolecules in normal,adenomatous,and carcinomatous colon:evidence for the neotransformation[J].Macromol biosci.2004,4(5):483-96.
5 Buisine M P,Desreumaux P,Debailleul V,et al.Abnormalities in mucin gene expression in Crohn's disease[J].Inflamm Bowel Dis.1999,5(1):24-32.
6 Moniaux N,Nollet S,Porchet N,et al.Complete sequence of the human mucin MUC4:a putative cell membrane-associated mucin[J].Biochem J,1999,338(Pt2):325-333.
7 Kanno A,Satoh K,Kimurak,et al.The expression of MUC4 and MUC5AC is related to the biologic malignancy of intraductal papillary mucinous neoplasms of the pancreas[J].Pancreas,2006,33(4):391-396.
8 Piessen G,Jonckheere N,Vincent A,et al.Regulation of the human mucin MUC4 by taurodeoxycholic and taurochenodeoxycholic bile acids in oesophageal cancer cells is mediated by hepatocyte nuclear factor lalpha [J]. Biochem J, 2007, 402 (1):81-91.
9 Hanaoka J, Kontani K, Sawai S, et al. Analysis of MUC4 mucin expression in lung carcinoma cells and its immunogenicity [J]. Cancer, 2001,92:2148-2157.
10 Guillem P, BilleretV, Buisine MP, et al. Mucin gene expression and cell differentiation in human normal, premalignant and malignant esophagus [J]. Int J Cancer, 2000, 88: 856-861.
11 Buisine MP, Devisme L, Degand P, et al. Developmental mucin gene expression in the gastroduodenal tract and accessory digestive glands II. Duodenum and liver, gallbladder, and pancreas. J Histochem Cytochem, 2000, 48: 1667-1676.
12 Weed DT, Gomez-Fernandez C, Bonfante E, et al. MUC4 (sialomucin complex) expression in salivary gland tumors and squamous cell carcinoma of the upper aerodigestive tract. Otolaryngol Head Neck Surg, 2001, 124: 127-141.
13 Carraway KL, Price-Schiavi SA, Komatsu M,et al. Muc4/Sialomucin complex in the mammary gland and breast cancer. J Mammary Gland Biol Neoplasia, 2001, 6: 323-337.
14 Singh AP, Moniaux N, Chauhan SC, et al. Inhibition of MUC4 expression suppresses pancreatic tumor cell growthand metastasis [J]. Cancer Res, 2004, 64 (2): 622- 630
15 Pardoll DM.Therapeutic vaccination for cancer[J].Clin Immunol 2000;95:S44-62.
16 Salskov-Iversen M,Berger CL,Edelson RL,et al.Rapid construction of a dendritic cell vaccine through physical perturbation and apoptotic malignant T cell loading.J Immune Based Ther Vaccines,2005,3(4):8518
17 陈建敏,高文涛,苗毅,等.MUC4 HLA-A*0201限制性CTL表位的预测及结合力分析[J].南京医科大学学报,2006,26(7):513-515.
18 Ahmadzadeh M,Rosenberg SA.TGF-beta I attenuates the acquisition and expression of effecter function by tumor antigen-specific human memory CD8 T cells[J].J Immunol,2005,174(9):5215-5223.
19 罗军,靳风烁.肿瘤的免疫逃逸机制研究进展[J].肿瘤防治研究,2004,3 1(7):454-456.
20 韩超峰,曹雪涛.肿瘤诱导的树突状细胞功能缺陷及机制[J].中国肿瘤生物治疗杂志,2005,12(2):89-93.
21 Bodey B,Siegel SE,Kaiser HE,et al.Antigen presentation by dendritic cells and their significance in antine-oplastic immunotherapy[J].In Vivo,2004,18(1):81-100.
22 Yang L,Carbone DP.Tumor-host immune interactions and dendritic cell dysfunction[J].Adv Cancer Res,2004,92(7):13-27.
23 Liu YJ.Dendritic cell subsets and lineages,and their functions in innate and adaptive immnity[J].J Immunol,2001,106:259-269.
24 陈巍,蔡莉.树突状细胞治疗肿瘤的研究现状[J].实用肿瘤学杂志,2007,21(3):293-295
25 Li B,Wang Y,Chen J,et al.Identification of a new HLA-A~*0201-restricted CD8~+ T cell epitope from hepatocellular carcinoma-associated antigen HCA587[J].Clin Exp Immunol,2005,140(2):310-319.
26 Groeper C,Gambazzi F,Zajac P,et al.Cancer/testis antigen expression and specific cytotoxic T lymphocyte responses in non small cell lung cancer[J].Int J Cancer.2007,120(2):337-343.
27 Pang PH,Chan KT,Tse LY,et al.Induction of cytotoxic T cell response against HCA661 positive cancer cells through activation with novel HLA-A ~*0201 restricted epitopes.Cancer Lett[J].2007,256(2):178-185.
28 Grover A,Kim GJ,Lizee G,et al.Intralymphatic dendritic cell vaccination induces tumor antigen-specific,skin-homing T lymphocytes[J].Clin Cancer Res.2006,12(19):5801-5808.
29 Parmiani G,Castelli C,Dalerba P,et al.Cancer immunotherapy with peptide-based vaccines:what have we achieved? Where are we going[J]? J Natl Cancer Inst,2002,94(11):805-818
30 Han JF,Zhao TT,Liu HL,et al.Identification of a new HLA-A~*0201-restricted cytotoxic T lymphocyte epitope from CML28[J].Cancer Immunol Immunother.2006,55(12):1575-1583.
1. Banchereau J, Briere C, Davoust S, etal. Immunobilogy of dendritic cells [J]. Annu Rev Immunol, 2000: 767-811.
2. Xie YA, Luo XL, Liang AM, etal. In vitro induction of anti-tumor responsiveness by dendritic cells pulsen with hepaocarcinoma cells antigen [J].World Chinese Digest logy, 2004, 12(6): 1479-1481.
3. Heufler C , Koch F , Stanz 1U , et al. Interleukin-12 is produced by dendritic cells and mediates T helper 1 development as well as interferon-gamma production by T helper 1 cells [J]. Eur J Immunol, 1996,26(3):659-668.
4. Norbury CC. Dendritic Cells Pulsed with RNA are Potent Antigen-presenting Cells in Vitro and in Vivo [J]. Eur J Immunol, 1997, 27(1):280-288.
5. Zhou LJ, Tedder TF. A Distinct Pattern of cytokine Gene Expression by Human CD~+ 83 Blood Dendritic Cells [J]. Blood, 1995, 86 (9): 3295-3301.
6. Fong L, Engleman EG. Dendric Cells in Cancer Immuno Therapy [J]. Annu Rev Immunol, 2000, 18: 245-273.
7. Chang E Y, Chen C H, Ji H, et al. Antigen-specific cancer immunotherapy using a GM-CSF secreting allogeneic tumor cell-based vaccine [J]. Int J Cancer, 2000, 86(5): 725-730.
8. Chong H, Hutchinson G, Hart IR, et al. Expression of B7 co-stimulatory molecules by B16 melanoma results in a naturalkiller cell dependent systemic immuneity only against B7 expressing tumors[J].Br J Cancer,1998,78(8):1043-1050.
9.Nestle FO,Burg G,Dummer R,et al.New perspectives on Immunobiology and immuneotherapy of melanoma[J].Immunlogy Today,1999,20(1):5-7.
10.Ugurel S,Seiter S,Rappl G,et al.Heterogenous susceptibility to CD95-induced apoptosis in melanoma cells correlates with bcl-2 and bcl-x expression and is sensitive to modulation by interferon-γ[J].Int J Cancer,1999,82(5):727-736.
11.Brossart P,Wirths S,Stuhler G,et al.Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptidepulsed dendritic cells[J].Blood,2000;96(9):3102-3108.
12.Nestle FO,Alijagic S,Gilliet M.Vaccination of melanoma patients with peptide or tumor lysate pulsed dendritic cells[J].Nat Med,1998;4(3):328-332.
13.朱学军,曹雪涛,雷虹等.弱酸洗脱提取的肿瘤抗原肽致敏的树突状细胞对CTL的体外激活效应[J].中华微生物学和免疫学杂志,2000;20(2):98-100.
14.Gabrilovich D I,Cunningham H T,Carbone D P.IL-12 and mutant p53 peptide-pulsed dendritic cells for the specific immunotherapy of cancer[J].J Immunother Emphasis Tumor Immunol,1996;19(6):414-418.
15.Strome SE,Voss S,Wilcox R,et al.Strategies for antigen loading of dendritic cells to enhance the antitumor immune response[J]. Cancer Res, 2002, 62(6): 1884-1889.
16. Zitvogel L ,Fernandez N ,Lozier A , et al . Dendritic cells or their exosomes are effective biotherapies of cancer [J]. Eur J Cancer, 1999,35(Suppl 3):36-38.
17. Heiser A, Coleman D, Dannull J, et al. Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors[J]. J Clin Invest, 2002 Feb; 109 (3): 409-417.
18. Choudhury BA, Liang JC, Thomas EK, et al. Dendritic cells derived in vitro from acute myelogenous leukemia cells stimulate autologous, antileukemic T-cell responses [J] Blood, 1999, 93 (3): 780-786
19. Gong J, Chen D, Kashiwaba M, et al. Reversal of tolerance to human MUC1 antigen in MUC1 transgenic mice immunized with fusions of dendritic and carcinoma cells [J]. Proc Natl Acad Sci USA, 1998, 95 (11): 6279-6283.
20. Gong J, Avigan D, Chen D, et al. Activation of antitumor cytotoxic T lymphocytes by fusions of human dendritic cells and breast carcinoma cells [J]. Proc Natl Acad Sci USA, 2000, 97(6): 2715-2718.
2 汪荣泉,房殿春,刘为纹,等.用BC2单抗检测正常腺上皮组织中表达MUC1基因的表达[J].第三军医大学学报,1999,21(6):413-415.
3 汪荣泉,房殿春,罗元辉,等.胃癌及癌前病变组织中MUC5AC基因的异常表达及意义[J].癌症,2000,19(2):121-123.
4 Aksoy N,Akinci O F.Mucin macromolecules in normal,adenomatous,and carcinomatous colon:evidence for the neotransformation[J].Macromol biosci.2004,4(5):483-96.
5 Buisine M P,Desreumaux P,Debailleul V,et al.Abnormalities in mucin gene expression in Crohn's disease[J].Inflamm Bowel Dis.1999,5(1):24-32.
6 Moniaux N,Nollet S,Porchet N,et al.Complete sequence of the human mucin MUC4:a putative cell membrane-associated mucin[J].Biochem J,1999,338(Pt2):325-333.
7 Kanno A,Satoh K,Kimurak,et al.The expression of MUC4 and MUC5AC is related to the biologic malignancy of intraductal papillary mucinous neoplasms of the pancreas[J].Pancreas,2006,33(4):391-396.
8 Piessen G,Jonckheere N,Vincent A,et al.Regulation of the human mucin MUC4 by taurodeoxycholic and taurochenodeoxycholic bile acids in oesophageal cancer cells is mediated by hepatocyte nuclear factor lalpha [J]. Biochem J, 2007, 402 (1):81-91.
9 Hanaoka J, Kontani K, Sawai S, et al. Analysis of MUC4 mucin expression in lung carcinoma cells and its immunogenicity [J]. Cancer, 2001,92:2148-2157.
10 Guillem P, BilleretV, Buisine MP, et al. Mucin gene expression and cell differentiation in human normal, premalignant and malignant esophagus [J]. Int J Cancer, 2000, 88: 856-861.
11 Buisine MP, Devisme L, Degand P, et al. Developmental mucin gene expression in the gastroduodenal tract and accessory digestive glands II. Duodenum and liver, gallbladder, and pancreas. J Histochem Cytochem, 2000, 48: 1667-1676.
12 Weed DT, Gomez-Fernandez C, Bonfante E, et al. MUC4 (sialomucin complex) expression in salivary gland tumors and squamous cell carcinoma of the upper aerodigestive tract. Otolaryngol Head Neck Surg, 2001, 124: 127-141.
13 Carraway KL, Price-Schiavi SA, Komatsu M,et al. Muc4/Sialomucin complex in the mammary gland and breast cancer. J Mammary Gland Biol Neoplasia, 2001, 6: 323-337.
14 Singh AP, Moniaux N, Chauhan SC, et al. Inhibition of MUC4 expression suppresses pancreatic tumor cell growthand metastasis [J]. Cancer Res, 2004, 64 (2): 622- 630
15 Pardoll DM.Therapeutic vaccination for cancer[J].Clin Immunol 2000;95:S44-62.
16 Salskov-Iversen M,Berger CL,Edelson RL,et al.Rapid construction of a dendritic cell vaccine through physical perturbation and apoptotic malignant T cell loading.J Immune Based Ther Vaccines,2005,3(4):8518
17 陈建敏,高文涛,苗毅,等.MUC4 HLA-A*0201限制性CTL表位的预测及结合力分析[J].南京医科大学学报,2006,26(7):513-515.
18 Ahmadzadeh M,Rosenberg SA.TGF-beta I attenuates the acquisition and expression of effecter function by tumor antigen-specific human memory CD8 T cells[J].J Immunol,2005,174(9):5215-5223.
19 罗军,靳风烁.肿瘤的免疫逃逸机制研究进展[J].肿瘤防治研究,2004,3 1(7):454-456.
20 韩超峰,曹雪涛.肿瘤诱导的树突状细胞功能缺陷及机制[J].中国肿瘤生物治疗杂志,2005,12(2):89-93.
21 Bodey B,Siegel SE,Kaiser HE,et al.Antigen presentation by dendritic cells and their significance in antine-oplastic immunotherapy[J].In Vivo,2004,18(1):81-100.
22 Yang L,Carbone DP.Tumor-host immune interactions and dendritic cell dysfunction[J].Adv Cancer Res,2004,92(7):13-27.
23 Liu YJ.Dendritic cell subsets and lineages,and their functions in innate and adaptive immnity[J].J Immunol,2001,106:259-269.
24 陈巍,蔡莉.树突状细胞治疗肿瘤的研究现状[J].实用肿瘤学杂志,2007,21(3):293-295
25 Li B,Wang Y,Chen J,et al.Identification of a new HLA-A~*0201-restricted CD8~+ T cell epitope from hepatocellular carcinoma-associated antigen HCA587[J].Clin Exp Immunol,2005,140(2):310-319.
26 Groeper C,Gambazzi F,Zajac P,et al.Cancer/testis antigen expression and specific cytotoxic T lymphocyte responses in non small cell lung cancer[J].Int J Cancer.2007,120(2):337-343.
27 Pang PH,Chan KT,Tse LY,et al.Induction of cytotoxic T cell response against HCA661 positive cancer cells through activation with novel HLA-A ~*0201 restricted epitopes.Cancer Lett[J].2007,256(2):178-185.
28 Grover A,Kim GJ,Lizee G,et al.Intralymphatic dendritic cell vaccination induces tumor antigen-specific,skin-homing T lymphocytes[J].Clin Cancer Res.2006,12(19):5801-5808.
29 Parmiani G,Castelli C,Dalerba P,et al.Cancer immunotherapy with peptide-based vaccines:what have we achieved? Where are we going[J]? J Natl Cancer Inst,2002,94(11):805-818
30 Han JF,Zhao TT,Liu HL,et al.Identification of a new HLA-A~*0201-restricted cytotoxic T lymphocyte epitope from CML28[J].Cancer Immunol Immunother.2006,55(12):1575-1583.
1. Banchereau J, Briere C, Davoust S, etal. Immunobilogy of dendritic cells [J]. Annu Rev Immunol, 2000: 767-811.
2. Xie YA, Luo XL, Liang AM, etal. In vitro induction of anti-tumor responsiveness by dendritic cells pulsen with hepaocarcinoma cells antigen [J].World Chinese Digest logy, 2004, 12(6): 1479-1481.
3. Heufler C , Koch F , Stanz 1U , et al. Interleukin-12 is produced by dendritic cells and mediates T helper 1 development as well as interferon-gamma production by T helper 1 cells [J]. Eur J Immunol, 1996,26(3):659-668.
4. Norbury CC. Dendritic Cells Pulsed with RNA are Potent Antigen-presenting Cells in Vitro and in Vivo [J]. Eur J Immunol, 1997, 27(1):280-288.
5. Zhou LJ, Tedder TF. A Distinct Pattern of cytokine Gene Expression by Human CD~+ 83 Blood Dendritic Cells [J]. Blood, 1995, 86 (9): 3295-3301.
6. Fong L, Engleman EG. Dendric Cells in Cancer Immuno Therapy [J]. Annu Rev Immunol, 2000, 18: 245-273.
7. Chang E Y, Chen C H, Ji H, et al. Antigen-specific cancer immunotherapy using a GM-CSF secreting allogeneic tumor cell-based vaccine [J]. Int J Cancer, 2000, 86(5): 725-730.
8. Chong H, Hutchinson G, Hart IR, et al. Expression of B7 co-stimulatory molecules by B16 melanoma results in a naturalkiller cell dependent systemic immuneity only against B7 expressing tumors[J].Br J Cancer,1998,78(8):1043-1050.
9.Nestle FO,Burg G,Dummer R,et al.New perspectives on Immunobiology and immuneotherapy of melanoma[J].Immunlogy Today,1999,20(1):5-7.
10.Ugurel S,Seiter S,Rappl G,et al.Heterogenous susceptibility to CD95-induced apoptosis in melanoma cells correlates with bcl-2 and bcl-x expression and is sensitive to modulation by interferon-γ[J].Int J Cancer,1999,82(5):727-736.
11.Brossart P,Wirths S,Stuhler G,et al.Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptidepulsed dendritic cells[J].Blood,2000;96(9):3102-3108.
12.Nestle FO,Alijagic S,Gilliet M.Vaccination of melanoma patients with peptide or tumor lysate pulsed dendritic cells[J].Nat Med,1998;4(3):328-332.
13.朱学军,曹雪涛,雷虹等.弱酸洗脱提取的肿瘤抗原肽致敏的树突状细胞对CTL的体外激活效应[J].中华微生物学和免疫学杂志,2000;20(2):98-100.
14.Gabrilovich D I,Cunningham H T,Carbone D P.IL-12 and mutant p53 peptide-pulsed dendritic cells for the specific immunotherapy of cancer[J].J Immunother Emphasis Tumor Immunol,1996;19(6):414-418.
15.Strome SE,Voss S,Wilcox R,et al.Strategies for antigen loading of dendritic cells to enhance the antitumor immune response[J]. Cancer Res, 2002, 62(6): 1884-1889.
16. Zitvogel L ,Fernandez N ,Lozier A , et al . Dendritic cells or their exosomes are effective biotherapies of cancer [J]. Eur J Cancer, 1999,35(Suppl 3):36-38.
17. Heiser A, Coleman D, Dannull J, et al. Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors[J]. J Clin Invest, 2002 Feb; 109 (3): 409-417.
18. Choudhury BA, Liang JC, Thomas EK, et al. Dendritic cells derived in vitro from acute myelogenous leukemia cells stimulate autologous, antileukemic T-cell responses [J] Blood, 1999, 93 (3): 780-786
19. Gong J, Chen D, Kashiwaba M, et al. Reversal of tolerance to human MUC1 antigen in MUC1 transgenic mice immunized with fusions of dendritic and carcinoma cells [J]. Proc Natl Acad Sci USA, 1998, 95 (11): 6279-6283.
20. Gong J, Avigan D, Chen D, et al. Activation of antitumor cytotoxic T lymphocytes by fusions of human dendritic cells and breast carcinoma cells [J]. Proc Natl Acad Sci USA, 2000, 97(6): 2715-2718.