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研究生:冉毅驊
研究生(外文):Jan Yi-Hua
論文名稱:在小鼠建立攝護腺癌樹突狀細胞腫瘤疫苗
論文名稱(外文):Establishment of Dendritic Cell-Based Tumor Vaccine for Prostate Cancer in Mice
指導教授:葉世榮葉世榮引用關係江啟勳
指導教授(外文):Yeh S-RChiang Chi-Shiun
學位類別:碩士
校院名稱:國立清華大學
系所名稱:分子醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:英文
論文頁數:40
中文關鍵詞:骨髓樹突狀細胞樹突狀細胞腫瘤疫苗攝護腺癌放射線
外文關鍵詞:Bone marrow-derived dendritic cellsDendritic cells tumor vaccineProstate cancerRadiation
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利用樹突狀細胞(Dendritic cells)呈現腫瘤相關的抗原能夠有效地誘發對抗腫瘤的免疫力。當無法得知特定腫瘤抗原的情況下,利用整個腫瘤細胞作為抗原來刺激樹突狀細胞是一個值得研究的策略。本篇論文的研究目的是在小鼠的動物模式上建立攝護腺癌的樹突狀細胞腫瘤疫苗,並且評估以下三種刺激樹突狀細胞的策略:1. 以腫瘤細胞裂解物(Tumor lysates)刺激樹突狀細胞。2. 以放射線照射的腫瘤細胞刺激樹突狀細胞。3. 以放射線照射的腫瘤細胞加上細胞激素-3(Interleukin-3, IL-3)共同刺激樹突狀細胞。樹突狀細胞可經由骨髓細胞在GM-CSF和IL-4五至六天的培養下分化增生而得,LPS的加入則會刺激樹突狀細胞更進一步分化為成熟樹突狀細胞。這些在體外培養產生的骨髓樹突狀細胞具有典型的樹突狀型態、細胞表面特殊分子、吞噬能力以及刺激不同源T細胞增生的能力。在本篇研究中,以放射線照射的TRAMP C1細胞株為抗原刺激樹突狀細胞做為攝護腺癌腫瘤疫苗,在預防腫瘤產生以及減緩腫瘤生長上最為有效,而IL-3的存在並沒有進一步增強這樣的免疫力。以TRAMP C1 lysates刺激樹突狀細胞相較之下無法有效地誘發抗腫瘤的免疫力。從本篇的實驗結果認為,使用放射線照射的腫瘤細胞做為抗原,是發展攝護腺癌樹突狀細胞腫瘤疫苗很有效的方式。
Dendritic cells(DCs)pulsed with tumor-associated antigens can effectively induce anti-tumor immunity. Antigen loading of DC with whole tumor cell preparations is an attractive mean when defined tumor-associated antigens are not available. The goal of this study was to establish a DC-based vaccine model for prostate cancer in mice and to evaluate three antigen loading strategies: DCs pulsed with whole tumor lysates, DCs pulsed with irradiated tumor cells, and DCs pulsed with irradiated tumor cells in the presence of IL-3. Mouse bone marrow-derived dendritic cells(BM-DCs) were generated after 5-6 days culture with GM-CSF and IL-4. Further maturation of BM-DCs could be induced by LPS for 24 hours. In vitro generated BM-DCs displayed typical DC morphology, surface markers expression, phagocytosis capacity and allostimulatory ability of T cell proliferation. DCs pulsed with irradiated TRAMP C1 cells were most effective in preventing or delaying tumor growth on tumor challenge. The presence of IL-3 did not further enhance this protective immunity. DCs pulsed with TRAMP C1 lystates seemed to be less effective compared to irradiated TRAMP C1 cells. These data suggested DCs pulsed with irradiated tumor cells is the most promising strategy in developing DC-based tumor vaccine for prostate cancer.
中文摘要 I
ABSTRACT II
誌謝 III
CONTENTS IV
INTRODUCTION 1
DCs subtypes 1
DC maturation 2
Antigen uptake, processing and presentation 3
Manipulation of DCs for cancer immunotherapy 4
MATERIALS AND METHODS 5
Animals 5
Cell lines and culture condition 5
Antibodies and reagents 6
Epitope prediction and peptide synthesis 6
Murine bone marrow-derived dendritic cells 7
DC phenotyping by FACS 7
Phagocytosis assay 8
XTT cell proliferation assay 8
Allogenic mixed lymphocyte reaction 9
Apoptosis detection after radiation 9
Phagocytosis of irradiated TRAMP C1 10
DC vaccine preparation 10
Immunization of mice and tumor challenge in vivo 10
CTL assay 11
RESULTS 12
Generation of bone marrow-derived dendritic cells 12
Surface markers phenotyping 13
Phagocytosis assay and allogenic mixed lymphocyte reaction 13
Irradiated TRAMP C1 cells are phagocytosed by DCs 14
DCs pulsed with irradiated TRAMP C1 cells up-regulate CD80, CD86 and MHC II expression 15
Induction of protective immunity by DCs pulsed with irradiated TRAMP C1 cells 16
Tumor specific CTL responses induced by DCs pulsed with irradiated TRAMP C1 cells 17
DISCUSSION 19
FIGURE LEGENDS 24
FIGURES 28
REFERENCES 37
1. Steinman, RM The dendritic cell system and its role in immunogenicity. Annu Rev Immunol, 1991; 9(271-296.
2. Shortman, K and Liu, YJ Mouse and human dendritic cell subtypes. Nat Rev Immunol, 2002; 2(3): 151-161.
3. Shortman, K and Wu, L Parentage and heritage of dendritic cells. Blood, 2001; 97(11): 3325.
4. den Haan, JM, Lehar, SM, and Bevan, MJ CD8(+) but not CD8(-) dendritic cells cross-prime cytotoxic T cells in vivo. J Exp Med, 2000; 192(12): 1685-1696.
5. Pooley, JL, Heath, WR, and Shortman, K Cutting edge: intravenous soluble antigen is presented to CD4 T cells by CD8- dendritic cells, but cross-presented to CD8 T cells by CD8+ dendritic cells. J Immunol, 2001; 166(9): 5327-5330.
6. Reis e Sousa, C and Germain, RN Analysis of adjuvant function by direct visualization of antigen presentation in vivo: endotoxin promotes accumulation of antigen-bearing dendritic cells in the T cell areas of lymphoid tissue. J Immunol, 1999; 162(11): 6552-6561.
7. Pulendran, B, Smith, JL, Caspary, G, et al. Distinct dendritic cell subsets differentially regulate the class of immune response in vivo. Proc Natl Acad Sci U S A, 1999; 96(3): 1036-1041.
8. Smith, AL and Fazekas de St Groth, B Antigen-pulsed CD8alpha+ dendritic cells generate an immune response after subcutaneous injection without homing to the draining lymph node. J Exp Med, 1999; 189(3): 593-598.
9. Aderem, A and Ulevitch, RJ Toll-like receptors in the induction of the innate immune response. Nature, 2000; 406(6797): 782-787.
10. Kalinski, P, Hilkens, CM, Wierenga, EA, and Kapsenberg, ML T-cell priming by type-1 and type-2 polarized dendritic cells: the concept of a third signal. Immunol Today, 1999; 20(12): 561-567.
11. Regnault, A, Lankar, D, Lacabanne, V, et al. Fcgamma receptor-mediated induction of dendritic cell maturation and major histocompatibility complex class I-restricted antigen presentation after immune complex internalization. J Exp Med, 1999; 189(2): 371-380.
12. Rescigno, M, Piguet, V, Valzasina, B, et al. Fas engagement induces the maturation of dendritic cells (DCs), the release of interleukin (IL)-1beta, and the production of interferon gamma in the absence of IL-12 during DC-T cell cognate interaction: a new role for Fas ligand in inflammatory responses. J Exp Med, 2000; 192(11): 1661-1668.
13. Ackerman, AL, Kyritsis, C, Tampe, R, and Cresswell, P Early phagosomes in dendritic cells form a cellular compartment sufficient for cross presentation of exogenous antigens. Proc Natl Acad Sci U S A, 2003; 100(22): 12889-12894.
14. Fonteneau, JF, Larsson, M, and Bhardwaj, N Interactions between dead cells and dendritic cells in the induction of antiviral CTL responses. Curr Opin Immunol, 2002; 14(4): 471-477.
15. Guermonprez, P, Saveanu, L, Kleijmeer, M, Davoust, J, Van Endert, P, and Amigorena, S ER-phagosome fusion defines an MHC class I cross-presentation compartment in dendritic cells. Nature, 2003; 425(6956): 397-402.
16. Steinman, RM and Dhodapkar, M Active immunization against cancer with dendritic cells: the near future. Int J Cancer, 2001; 94(4): 459-473.
17. Ashley, DM, Faiola, B, Nair, S, Hale, LP, Bigner, DD, and Gilboa, E Bone marrow-generated dendritic cells pulsed with tumor extracts or tumor RNA induce antitumor immunity against central nervous system tumors. J Exp Med, 1997; 186(7): 1177-1182.
18. Fields, RC, Shimizu, K, and Mule, JJ Murine dendritic cells pulsed with whole tumor lysates mediate potent antitumor immune responses in vitro and in vivo. Proc Natl Acad Sci U S A, 1998; 95(16): 9482-9487.
19. Galea-Lauri, J, Darling, D, Mufti, G, Harrison, P, and Farzaneh, F Eliciting cytotoxic T lymphocytes against acute myeloid leukemia-derived antigens: evaluation of dendritic cell-leukemia cell hybrids and other antigen-loading strategies for dendritic cell-based vaccination. Cancer Immunol Immunother, 2002; 51(6): 299-310.
20. 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.
21. Wolfers, J, Lozier, A, Raposo, G, et al. Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming. Nat Med, 2001; 7(3): 297-303.
22. Foster, BA, Gingrich, JR, Kwon, ED, Madias, C, and Greenberg, NM Characterization of prostatic epithelial cell lines derived from transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Cancer Res, 1997; 57(16): 3325-3330.
23. Lu, J and Celis, E Use of two predictive algorithms of the world wide web for the identification of tumor-reactive T-cell epitopes. Cancer Res, 2000; 60(18): 5223-5227.
24. Inaba, K, Inaba, M, Romani, N, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med, 1992; 176(6): 1693-1702.
25. Pulaski, BA, Yeh, KY, Shastri, N, et al. Interleukin 3 enhances cytotoxic T lymphocyte development and class I major histocompatibility complex "re-presentation" of exogenous antigen by tumor-infiltrating antigen-presenting cells. Proc Natl Acad Sci U S A, 1996; 93(8): 3669-3674.
26. Yeh, KY, McAdam, AJ, Pulaski, BA, Shastri, N, Frelinger, JG, and Lord, EM IL-3 enhances both presentation of exogenous particulate antigen in association with class I major histocompatibility antigen and generation of primary tumor-specific cytolytic T lymphocytes. J Immunol, 1998; 160(12): 5773-5780.
27. Liu, L, Chahroudi, A, Silvestri, G, et al. Visualization and quantification of T cell-mediated cytotoxicity using cell-permeable fluorogenic caspase substrates. Nat Med, 2002; 8(2): 185-189.
28. Lutz, MB, Kukutsch, N, Ogilvie, AL, et al. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods, 1999; 223(1): 77-92.
29. Jiang, W, Swiggard, WJ, Heufler, C, et al. The receptor DEC-205 expressed by dendritic cells and thymic epithelial cells is involved in antigen processing. Nature, 1995; 375(6527): 151-155.
30. Goldszmid, RS, Idoyaga, J, Bravo, AI, Steinman, R, Mordoh, J, and Wainstok, R Dendritic cells charged with apoptotic tumor cells induce long-lived protective CD4+ and CD8+ T cell immunity against B16 melanoma. J Immunol, 2003; 171(11): 5940-5947.
31. Sauter, B, Albert, ML, Francisco, L, Larsson, M, Somersan, S, and Bhardwaj, N Consequences of cell death: exposure to necrotic tumor cells, but not primary tissue cells or apoptotic cells, induces the maturation of immunostimulatory dendritic cells. J Exp Med, 2000; 191(3): 423-434.
32. Paczesny, S, Beranger, S, Salzmann, JL, Klatzmann, D, and Colombo, BM Protection of mice against leukemia after vaccination with bone marrow-derived dendritic cells loaded with apoptotic leukemia cells. Cancer Res, 2001; 61(6): 2386-2389.
33. Ronchetti, A, Rovere, P, Iezzi, G, et al. Immunogenicity of apoptotic cells in vivo: role of antigen load, antigen-presenting cells, and cytokines. J Immunol, 1999; 163(1): 130-136.
34. Matzinger, P Tolerance, danger, and the extended family. Annu Rev Immunol, 1994; 12(991-1045.
35. Gallucci, S and Matzinger, P Danger signals: SOS to the immune system. Curr Opin Immunol, 2001; 13(1): 114-119.
36. Basu, S, Binder, RJ, Suto, R, Anderson, KM, and Srivastava, PK Necrotic but not apoptotic cell death releases heat shock proteins, which deliver a partial maturation signal to dendritic cells and activate the NF-kappa B pathway. Int Immunol, 2000; 12(11): 1539-1546.
37. Bogdan, C The function of type I interferons in antimicrobial immunity. Curr Opin Immunol, 2000; 12(4): 419-424.
38. Caux, C, Massacrier, C, Vanbervliet, B, et al. Activation of human dendritic cells through CD40 cross-linking. J Exp Med, 1994; 180(4): 1263-1272.
39. Sallusto, F, Cella, M, Danieli, C, and Lanzavecchia, A Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J Exp Med, 1995; 182(2): 389-400.
40. Zepter, K, Haffner, A, Soohoo, LF, et al. Induction of biologically active IL-1 beta-converting enzyme and mature IL-1 beta in human keratinocytes by inflammatory and immunologic stimuli. J Immunol, 1997; 159(12): 6203-6208.
41. Shi, Y, Evans, JE, and Rock, KL Molecular identification of a danger signal that alerts the immune system to dying cells. Nature, 2003; 425(6957): 516-521.
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