黑色素瘤相关抗原基因及其抗原肽负载树突状细胞免疫治疗乳腺癌的实验研究
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摘要
背景 自1991年发现第一个黑色素瘤相关抗原基因(Melanoma-associated Antigen Gene,Mage)及其抗原肽(MZ2-E)以来,多年来,人们已在不同肿瘤组织中,发现多个Mage基因,均定位于X染色体,有着一定的同源性,组成了一个Mage基因家族,分为A、B、C、D、E、F六个亚家族。进一步的研究发现,其中A、B、C三个亚家族存在一些独特的特征,只在肿瘤、睾丸和胎盘组织中表达,在其它正常组织迄今为止还没有发现其表达。而且其表达产物是一种肿瘤排斥性抗原,细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)可以识别该抗原。这提示Mage基因有可能作为肿瘤相关的检测标志,有助于肿瘤的诊断与治疗,而其抗原有可能做为肿瘤免疫治疗的靶抗原进行免疫治疗。
树突状细胞(dendritic cells,DCs)是一类形态独特的MHC携带细胞,有强有力的免疫刺激力和抗原递呈能力,也是唯一能激活初始型T细胞、诱发初次免疫应答的抗原递呈细胞。越来越多的证据表明,DC诱发的细胞免疫尤其是CTL介导的免疫应答,在消除恶性肿瘤中起着重要作用。利用树突状细胞负载抗原进行细胞过继免疫治疗(adoptive cellular immunotherapy,ACI)受到广泛关注。
基于Mage基因及其抗原肽的特征,我们对Mage基因在乳腺癌中的表达,在隐匿肿瘤细胞(occult tumor cells,OTC)检测中的价值,以
Background:Since van der Bruggen found the first melanoma-associated antigen gene (Mage) and its peptide MZ2-E in 1991, the researchers had found dozens of Mage in many different types of tumors. They were all mapped to chromosome X and shared certain homologous regions. They were all the members of Mage family, including six subgroups A、 B、 C、 D、 E、 F. And the subgroups A、 B. C shared certain homology. They has been found in many different types of malignant tumors, but not in normal adult tissues except the testis and placenta. Though scientists didin't know the physiological function of Mage genes encoding proteins, researchers had found that they were tumor antigens which could be recognized by specific T cells in malignant tumor. It suggeated us Mage maybe a novel marker to detect tumor and its peptide may be the most promising candidate for tumor-specific immunotherapy of cancer.Dendritic cells (DCs) were the most potent and the only antigen presenting cells (APC), which were capable of activating naive T cells and initiating primary immune response. Recently, more and more evidence showed that the cellular immunity induced by dendritic cells, especial the
CTL response, played an important role in the immune response against the malignant tumor. The adoptive cellular immunotherapy based on the dendritic cells loaded with antigen is becoming the highlight in the biology therpy of malignant tumor.The expression of Mage-A1 、 A2、 A3 in breast cancer tissues and four breast cancer cell lines was researched. Their mRNA was used as the marker to detecte the occult tumor cells in peripheral blood of patients with breast cancer. The experimental immunotherapy based on dendritic cells loaded with Mage-A3 peptide were studied on the mice model of breast cancer. 1. Expression of Mage-A1、 A2、 A3 in breast cancer tissue and breast cancer cell linesObjectives To study the expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines.Methods The expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines, MCF-7、 Sk-Br-3、 MDA-MB-435s and TM40D was detected by reverse transcription polymerase chain reaction (RT-PCR) . Results The positive expression rate of Mage-A in breast cancer tissues was 13/33 (39%) , of Mage-Al was 4/33 (12%) , of Mage-A2 was 8/33 ( 24%), and of Mage-A3 was 7/33( 21%), respectively. Both Mage-A1 and Mage-A3 were positive in breast cancer cell lines MCF-7 and Sk-Br-3. MDA-MB-435s expressed Mage-A2 and Mage-A3. Mage-A3 was positive in TM40D.
Conclusions Mage genes were often expressed in breast cancer, butexpression of Mages varied in the breast cancer cell lines. Mage geneencoding proteins are eligible for Mage-peptide-based activeimmunotherapy.2. Expression of Mage-A1、 A2、 A3 in the occult tumor cells in theperipheral blood of patients with breast cancerObjective To detect tumor cells in the peripheral blood of patients withbreast cancer by using the mRNA of the Mage-A1 and Mage-A3 genes asspecific tumor markers.Methods Peripheral blood was obtained from 40 breast cancer patients and20 patients with benign diseases. The mRNA of the Mage-A1 and Mage-A3genes in the peripheral blood mononuclear cells (PBMCs) was detected bynested RT-PCR. The Mage-A1 and Mage-A3 transcripts in the tumor tissuesof these breast cancer patients were also detected by RT-PCR.Results Of the 40 breast cancer patients, Mage-Al and Mage-A3 mRNAwere positive in 12.5% ( 5/40) and 17.5% ( 7/40 ) of PBMCs respectively, andin 15 % ( 6/40 ) and 22.5 % ( 9/ 40 ) of breast cancer tissues respectively. Inthe PBMCs of the 40 breast cancer patients, 10( 25 % )samples were detectedto express at least one type of Mage mRNA. Mage mRNA were not found inthe PBMCs from the patients whose tumors did not express the Mage genes,nor in the PBMCs from the 20 patients with benign diseases. The positiverate of Mage mRNA in the PBMCs was closely correlated with the TNM
stages.Conclusion Mage-A1 and Mage-A3 mRNA could be specificallydetected in the PBMCs of breast cancer pati
树突状细胞(dendritic cells,DCs)是一类形态独特的MHC携带细胞,有强有力的免疫刺激力和抗原递呈能力,也是唯一能激活初始型T细胞、诱发初次免疫应答的抗原递呈细胞。越来越多的证据表明,DC诱发的细胞免疫尤其是CTL介导的免疫应答,在消除恶性肿瘤中起着重要作用。利用树突状细胞负载抗原进行细胞过继免疫治疗(adoptive cellular immunotherapy,ACI)受到广泛关注。
基于Mage基因及其抗原肽的特征,我们对Mage基因在乳腺癌中的表达,在隐匿肿瘤细胞(occult tumor cells,OTC)检测中的价值,以
Background:Since van der Bruggen found the first melanoma-associated antigen gene (Mage) and its peptide MZ2-E in 1991, the researchers had found dozens of Mage in many different types of tumors. They were all mapped to chromosome X and shared certain homologous regions. They were all the members of Mage family, including six subgroups A、 B、 C、 D、 E、 F. And the subgroups A、 B. C shared certain homology. They has been found in many different types of malignant tumors, but not in normal adult tissues except the testis and placenta. Though scientists didin't know the physiological function of Mage genes encoding proteins, researchers had found that they were tumor antigens which could be recognized by specific T cells in malignant tumor. It suggeated us Mage maybe a novel marker to detect tumor and its peptide may be the most promising candidate for tumor-specific immunotherapy of cancer.Dendritic cells (DCs) were the most potent and the only antigen presenting cells (APC), which were capable of activating naive T cells and initiating primary immune response. Recently, more and more evidence showed that the cellular immunity induced by dendritic cells, especial the
CTL response, played an important role in the immune response against the malignant tumor. The adoptive cellular immunotherapy based on the dendritic cells loaded with antigen is becoming the highlight in the biology therpy of malignant tumor.The expression of Mage-A1 、 A2、 A3 in breast cancer tissues and four breast cancer cell lines was researched. Their mRNA was used as the marker to detecte the occult tumor cells in peripheral blood of patients with breast cancer. The experimental immunotherapy based on dendritic cells loaded with Mage-A3 peptide were studied on the mice model of breast cancer. 1. Expression of Mage-A1、 A2、 A3 in breast cancer tissue and breast cancer cell linesObjectives To study the expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines.Methods The expression of Mage-A1、 A2、 A3 in breast cancer tissues and four breast cancer cell lines, MCF-7、 Sk-Br-3、 MDA-MB-435s and TM40D was detected by reverse transcription polymerase chain reaction (RT-PCR) . Results The positive expression rate of Mage-A in breast cancer tissues was 13/33 (39%) , of Mage-Al was 4/33 (12%) , of Mage-A2 was 8/33 ( 24%), and of Mage-A3 was 7/33( 21%), respectively. Both Mage-A1 and Mage-A3 were positive in breast cancer cell lines MCF-7 and Sk-Br-3. MDA-MB-435s expressed Mage-A2 and Mage-A3. Mage-A3 was positive in TM40D.
Conclusions Mage genes were often expressed in breast cancer, butexpression of Mages varied in the breast cancer cell lines. Mage geneencoding proteins are eligible for Mage-peptide-based activeimmunotherapy.2. Expression of Mage-A1、 A2、 A3 in the occult tumor cells in theperipheral blood of patients with breast cancerObjective To detect tumor cells in the peripheral blood of patients withbreast cancer by using the mRNA of the Mage-A1 and Mage-A3 genes asspecific tumor markers.Methods Peripheral blood was obtained from 40 breast cancer patients and20 patients with benign diseases. The mRNA of the Mage-A1 and Mage-A3genes in the peripheral blood mononuclear cells (PBMCs) was detected bynested RT-PCR. The Mage-A1 and Mage-A3 transcripts in the tumor tissuesof these breast cancer patients were also detected by RT-PCR.Results Of the 40 breast cancer patients, Mage-Al and Mage-A3 mRNAwere positive in 12.5% ( 5/40) and 17.5% ( 7/40 ) of PBMCs respectively, andin 15 % ( 6/40 ) and 22.5 % ( 9/ 40 ) of breast cancer tissues respectively. Inthe PBMCs of the 40 breast cancer patients, 10( 25 % )samples were detectedto express at least one type of Mage mRNA. Mage mRNA were not found inthe PBMCs from the patients whose tumors did not express the Mage genes,nor in the PBMCs from the 20 patients with benign diseases. The positiverate of Mage mRNA in the PBMCs was closely correlated with the TNM
stages.Conclusion Mage-A1 and Mage-A3 mRNA could be specificallydetected in the PBMCs of breast cancer pati
引文
1. van der Bruggen P, Traversari C, Chomez P, et al. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science, 1991, 254(5038): 1643-1647.
2. Coulie PG, Weynants P, Lehmann F, et al. Genes coding for tumor antigens recognized by human cytolytic T lymphocytes. J Immunother, 1993, 14(2): 104-109
3. Scanlan MJ, Gure AO, Jungbluth AA, et al. Cancer/testis antigens: an expanding family of targets for cancer immunotherapy. Immunol Rev, 2002, 188: 22-32
4. Lucas S, De Smet C, Arden KC, et al. Identification of a new Mage gene with tumor-specific expression by representational difference analysis. Cancer Res, 1998, 58(4): 743-752
5. Kirkin AF, Dzhandzhugazyan KN, Zeuthen J. Cancer/testis antigens: structural and immunobiological properties. Cancer Invest, 2002, 20(2): 222-236
6. Muscatelli F, Walker AP, De Plaen E, et al. Isolation and characterization of a Mage gene family in the Xp21.3 region. Proc Natl Acad Sci U S A. 1995, 23, 92(11): 4987-1991
7. Lurquin C, De Smet C, Brasseur F, et al. Two members of the human MageB gene family located in Xp21.3 are expressed in tumors of various histological origins. Genomics. 1997, 46(3): 397-408
8. Lucas S, De Smet C, Arden KC, et al. Identification of a new Mage gene with tumor-specific expression by representational difference analysis. Cancer Res, 1998, 58(4): 743-752
9. Itoh K, Hayashi A, Nakao M, et al. Human tumor rejection antigens Mage. JBiochem, 1996, 119(3): 385-390
10. Stone B, Schummer M, Paley PJ, et al. Mage-F1, a novel ubiquitously expressed member of the Mage superfamily. Gene, 2001, 18, 267(2): 173-182
11. Kawano Y, Sasaki M, Nakahira K, et al. Structural characterization and chromosomal localization of the Mage-El gene. Gene, 2001 , 277(1-2): 129-137
12. Bodey B. Cancer-testis antigens: promising targets for antigen directed antineoplastic immunotherapy. Expert Opin Biol Ther, 2002, 2(6): 577-84
13. Juretic A, Spagnoli GC, Schultz TE, et al. Cancer/testis tumour-associated antigens : immunohistochemical detection with monoclonal antibodies. Lancet Oncol, 2003, 4(2): 104-109
14. Eichmuller S, Usener D, Thiel D, et al. Tumor-specific antigens in cutaneous T-cell lymphoma: expression and sero-reactivity. Int J Cancer, 2003, 104(4): 482-487
15. Kufer P, Zippelius A, Lutterbuse R, et al. Heterogeneous expression of Mage-A genes in occult disseminated tumor cells: a novel multimarker reverse transcription-polymerase chain reaction for diagnosis of micrometastatic disease. Cancer Res, 2002, 62(1): 251-261
16. Diel IJ , Kaufmann M, Costa SD, et al. Micrometastatic breast cancer cells in bone marrow at primary surgery: prognostic value in comparison with nodal status. J Natl Cancer Inst, 1996, 88(22): 1652-1664
17. Pierga JY, Bonneton C, Magdelenat H, et al. Clinical significance of proliferative potential of occult metastatic cells in bone marrow of patients with breast cancer. BrJ Cancer, 2003, 89(3): 539-545
18. Chung MA, Wazer D, Cady B. Contemporary management of bone marrow. Obstet Gynecol Clin North Am, 2002, 29(1): 173-188
19. Pantel K, Cote RJ, Fodstad O, et al. Detection and clinical importance of micrometastatic disease. J Natl Cancer Inst, 1999, 91(13): 1113-1124.
20. Pelkey TJ, Frierson HF, Bruns DE. Molecular and immunological detection of circulating tumor cells and micrometastases from solid tumors. Clinical Chem, 1996, 42(9): 1369-1381
21. 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
22. Hart-DN. Dendritic cells: unique leukocyte populations which control the primary immune response. Blood, 1997, 90(9): 3245-3287
23. Tarte K, Klein B. Dendritic cell-based vaccine: a promising approach for cancer immunotherapy. Leukemia, 1999, 13(5): 653-663
24. Stockwin LH, McGonagle D, Martin IG, et al. Dendritic cells: immunological sentinels with a central role in health and disease. Immunol Cell Biol, 2000, 78(2): 91-102
25. Dallal RM, Lotze MT. The dendritic cell and human cancer vaccines. Curr Opin Immunol, 2000,12(5): 583-588.
26. Sprinzl GM, Kacani L, Schrott-Fischer A, et al. Dendritic cell vaccines for cancer therapy. Cancer Treat Rev, 2001, 27(4): 247-255
27. Banchereau J, Schuler TB, Palucka AK, et al. Dendritic cells as vectors for therapy. Cell, 2001,106(3): 271-274
28. Thurner B, Haendle I, Roder C, et al. Vaccination with Mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med, 1999, 190(11): 1669-1678
29. Gunzer M, Janich S, Varga G, et al. Dendritic cells and tumor immunity. Semin Immunol, 2001, 13(5): 291-302
30. Burdin N, Moingeon P. Cancer vaccines based on dendritic cells loaded with tumor-associated antigens. Cell Biol Toxicol, 2001,17(2): 67-75
2. Coulie PG, Weynants P, Lehmann F, et al. Genes coding for tumor antigens recognized by human cytolytic T lymphocytes. J Immunother, 1993, 14(2): 104-109
3. Scanlan MJ, Gure AO, Jungbluth AA, et al. Cancer/testis antigens: an expanding family of targets for cancer immunotherapy. Immunol Rev, 2002, 188: 22-32
4. Lucas S, De Smet C, Arden KC, et al. Identification of a new Mage gene with tumor-specific expression by representational difference analysis. Cancer Res, 1998, 58(4): 743-752
5. Kirkin AF, Dzhandzhugazyan KN, Zeuthen J. Cancer/testis antigens: structural and immunobiological properties. Cancer Invest, 2002, 20(2): 222-236
6. Muscatelli F, Walker AP, De Plaen E, et al. Isolation and characterization of a Mage gene family in the Xp21.3 region. Proc Natl Acad Sci U S A. 1995, 23, 92(11): 4987-1991
7. Lurquin C, De Smet C, Brasseur F, et al. Two members of the human MageB gene family located in Xp21.3 are expressed in tumors of various histological origins. Genomics. 1997, 46(3): 397-408
8. Lucas S, De Smet C, Arden KC, et al. Identification of a new Mage gene with tumor-specific expression by representational difference analysis. Cancer Res, 1998, 58(4): 743-752
9. Itoh K, Hayashi A, Nakao M, et al. Human tumor rejection antigens Mage. JBiochem, 1996, 119(3): 385-390
10. Stone B, Schummer M, Paley PJ, et al. Mage-F1, a novel ubiquitously expressed member of the Mage superfamily. Gene, 2001, 18, 267(2): 173-182
11. Kawano Y, Sasaki M, Nakahira K, et al. Structural characterization and chromosomal localization of the Mage-El gene. Gene, 2001 , 277(1-2): 129-137
12. Bodey B. Cancer-testis antigens: promising targets for antigen directed antineoplastic immunotherapy. Expert Opin Biol Ther, 2002, 2(6): 577-84
13. Juretic A, Spagnoli GC, Schultz TE, et al. Cancer/testis tumour-associated antigens : immunohistochemical detection with monoclonal antibodies. Lancet Oncol, 2003, 4(2): 104-109
14. Eichmuller S, Usener D, Thiel D, et al. Tumor-specific antigens in cutaneous T-cell lymphoma: expression and sero-reactivity. Int J Cancer, 2003, 104(4): 482-487
15. Kufer P, Zippelius A, Lutterbuse R, et al. Heterogeneous expression of Mage-A genes in occult disseminated tumor cells: a novel multimarker reverse transcription-polymerase chain reaction for diagnosis of micrometastatic disease. Cancer Res, 2002, 62(1): 251-261
16. Diel IJ , Kaufmann M, Costa SD, et al. Micrometastatic breast cancer cells in bone marrow at primary surgery: prognostic value in comparison with nodal status. J Natl Cancer Inst, 1996, 88(22): 1652-1664
17. Pierga JY, Bonneton C, Magdelenat H, et al. Clinical significance of proliferative potential of occult metastatic cells in bone marrow of patients with breast cancer. BrJ Cancer, 2003, 89(3): 539-545
18. Chung MA, Wazer D, Cady B. Contemporary management of bone marrow. Obstet Gynecol Clin North Am, 2002, 29(1): 173-188
19. Pantel K, Cote RJ, Fodstad O, et al. Detection and clinical importance of micrometastatic disease. J Natl Cancer Inst, 1999, 91(13): 1113-1124.
20. Pelkey TJ, Frierson HF, Bruns DE. Molecular and immunological detection of circulating tumor cells and micrometastases from solid tumors. Clinical Chem, 1996, 42(9): 1369-1381
21. 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
22. Hart-DN. Dendritic cells: unique leukocyte populations which control the primary immune response. Blood, 1997, 90(9): 3245-3287
23. Tarte K, Klein B. Dendritic cell-based vaccine: a promising approach for cancer immunotherapy. Leukemia, 1999, 13(5): 653-663
24. Stockwin LH, McGonagle D, Martin IG, et al. Dendritic cells: immunological sentinels with a central role in health and disease. Immunol Cell Biol, 2000, 78(2): 91-102
25. Dallal RM, Lotze MT. The dendritic cell and human cancer vaccines. Curr Opin Immunol, 2000,12(5): 583-588.
26. Sprinzl GM, Kacani L, Schrott-Fischer A, et al. Dendritic cell vaccines for cancer therapy. Cancer Treat Rev, 2001, 27(4): 247-255
27. Banchereau J, Schuler TB, Palucka AK, et al. Dendritic cells as vectors for therapy. Cell, 2001,106(3): 271-274
28. Thurner B, Haendle I, Roder C, et al. Vaccination with Mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med, 1999, 190(11): 1669-1678
29. Gunzer M, Janich S, Varga G, et al. Dendritic cells and tumor immunity. Semin Immunol, 2001, 13(5): 291-302
30. Burdin N, Moingeon P. Cancer vaccines based on dendritic cells loaded with tumor-associated antigens. Cell Biol Toxicol, 2001,17(2): 67-75