脑膜瘤OY-TES-1mRNA表达及抗体血清检测
摘要
目的:初步了解OY-TES-1mRNA在脑膜瘤中的表达及血清抗体出现情况,探讨将其用于脑膜瘤免疫治疗及辅助诊断的可能性。
方法:收集病人脑膜瘤标本及配套血清30例,正常脑组织6例,正常人血清76例。提取总RNA,合成cDNA,采用逆转录-聚合酶链反应(RT-PCR)技术,扩增OY-TES-1开放阅读框架片段。以本实验室制备的OY-TES-1重组蛋白为抗原,用酶联免疫吸附分析技术(enzyme linkedimmunosorbent assay,ELISA)对病人及健康人血清OY-TES-1抗体进行检测。
结果:脑膜瘤中OY-TES-1mRNA阳性率为56.67%(17/30),6例正常脑组织中未能检测到OY-TES-1mRNA。ELISA检测发现,脑膜瘤患者血清OY-TES-1抗体阳性率为63.33%(19/30),正常人血清为阴性。在抗体阳性的病人中,女性13例,男性6例,阳性率分别为65%和60%。在检测的30例脑膜瘤中,40%(12例)为肿瘤组织mRNA和血清抗体均为阳性。
结论:OY-TES-1mRNA在脑膜瘤组织具有较高的阳性率,而在正常脑组织为阴性,提示OY-TES-1有望成为用于脑膜瘤免疫治疗的靶抗原。脑膜瘤患者血清具有较高OY-TES-1抗体阳性率,其临床意义有待进一步的确定。
Objective:To primarily examine the expression of OY-TES-1mRNA in meningioma tissue,and the occurrence of OY-TES-1 antibody in sera from patient with meningiomas.The opposibility of applying OY-TES-1 in tumor immunotherapy and assistant diagnosis is explored.
Methods:30 meningiomas tissues with patient's sera,6 normal brain tissues and 76 sera from health individual were collected.Exaction of total RNA was performed,and cDNA was synthesized.The fraction of opening reading frame of OY-TES-1 was amplifed with reverse-transcriptase polymerase chain reaction(RT-PCT)With OY-TES-1 recombinant protein produced by our laboratory as a antigen,enzyme linked immunosorbent assay(ELISA)was used to detect the OY-TES-1 antibody in serum from patient and health individual,
Results:The expression of OY-TES-1 mRNA in meningiomas was 56.67% (17/30).Normal brain tissues were all negative for the OY-TES-1 mRNA.The result of ELISA showed 63.33%(19/30)meningioma patients with serum antibody of OY-TES-1,while sera from health individual were all negative.Of 19 positive sera,female and male were 13(65%)and 6(60%),respectively.Of 30 samples tested,12(40%)was both mRNA in tissue and antibody in serum positive.
Conclusion:The expression of OY-TES-1 mRNA in meningiomas has higher frequence,while it was negative in the normal brain tissues tested,which suggested that OY-TES-1 would be a target antigen of immunotherapy for meningiomas.The clinical significance of OY-TES-1 antibody with higher frequence has to be further explored.
方法:收集病人脑膜瘤标本及配套血清30例,正常脑组织6例,正常人血清76例。提取总RNA,合成cDNA,采用逆转录-聚合酶链反应(RT-PCR)技术,扩增OY-TES-1开放阅读框架片段。以本实验室制备的OY-TES-1重组蛋白为抗原,用酶联免疫吸附分析技术(enzyme linkedimmunosorbent assay,ELISA)对病人及健康人血清OY-TES-1抗体进行检测。
结果:脑膜瘤中OY-TES-1mRNA阳性率为56.67%(17/30),6例正常脑组织中未能检测到OY-TES-1mRNA。ELISA检测发现,脑膜瘤患者血清OY-TES-1抗体阳性率为63.33%(19/30),正常人血清为阴性。在抗体阳性的病人中,女性13例,男性6例,阳性率分别为65%和60%。在检测的30例脑膜瘤中,40%(12例)为肿瘤组织mRNA和血清抗体均为阳性。
结论:OY-TES-1mRNA在脑膜瘤组织具有较高的阳性率,而在正常脑组织为阴性,提示OY-TES-1有望成为用于脑膜瘤免疫治疗的靶抗原。脑膜瘤患者血清具有较高OY-TES-1抗体阳性率,其临床意义有待进一步的确定。
Objective:To primarily examine the expression of OY-TES-1mRNA in meningioma tissue,and the occurrence of OY-TES-1 antibody in sera from patient with meningiomas.The opposibility of applying OY-TES-1 in tumor immunotherapy and assistant diagnosis is explored.
Methods:30 meningiomas tissues with patient's sera,6 normal brain tissues and 76 sera from health individual were collected.Exaction of total RNA was performed,and cDNA was synthesized.The fraction of opening reading frame of OY-TES-1 was amplifed with reverse-transcriptase polymerase chain reaction(RT-PCT)With OY-TES-1 recombinant protein produced by our laboratory as a antigen,enzyme linked immunosorbent assay(ELISA)was used to detect the OY-TES-1 antibody in serum from patient and health individual,
Results:The expression of OY-TES-1 mRNA in meningiomas was 56.67% (17/30).Normal brain tissues were all negative for the OY-TES-1 mRNA.The result of ELISA showed 63.33%(19/30)meningioma patients with serum antibody of OY-TES-1,while sera from health individual were all negative.Of 19 positive sera,female and male were 13(65%)and 6(60%),respectively.Of 30 samples tested,12(40%)was both mRNA in tissue and antibody in serum positive.
Conclusion:The expression of OY-TES-1 mRNA in meningiomas has higher frequence,while it was negative in the normal brain tissues tested,which suggested that OY-TES-1 would be a target antigen of immunotherapy for meningiomas.The clinical significance of OY-TES-1 antibody with higher frequence has to be further explored.
引文
1忠诚。神经外科学.武汉:湖北科学技术出版社,1998:512.
2黄文清.神经肿瘤病理学(第二版).军事医学科学出版社,北京,2001,479-506.
3Sawamura Y,de Tribolet N.Immunobiology of brain tumor.Adv Tech Stand Neurosurg,1990,17:3-64.
4Pollack IF,Okada H,Chambers WH.Exploitation of immune mechanisms in the treatment of central nervous system cancer.Semin Pediatr Neurol,2000,7(2):131-143.
5Pallasch CP,Struss AK,Munnia A,et al.Autoantibodies against GLEA2and PHF3 in glioblastoma:tumor-associated autoantibodies correlated with prolonged survival.Int J Cancer.2005,117(3):456-9.
6Liu G,Ying H,Zeng G,et al.HER-2,gpl00,and MAGE-1 are expressed in human glioblastoma and recognized by cytotoxic T cells.Cancer Res,2004,64(14):4980-4986.
7Rodolfo M,Luksch R,Stockert E,et al.Antigen-specific immunity in neuroblastoma patients:antibody and T-cell recognition of NY-ESO-1tumor antigen.Cancer Res,2003,63(20):6948-6955.
8Chen YT,Scanlan MJ,Sahin U,et al.A testicular antigen aberrantly expressed in human cancers detected by autologous antibody screening.Proc Natl Acad Sci U S A,1997,94:1914.
9Ono T,Kurashige T,Harada N,et al.Identification of proacrosin binding protein sp32 precursor as a human cancer/testis antigen.Proc Natl Acad Sci U S A,2001,98:3282.
10Okumura H,Noguchi Y,Uenaka A,et al.Identification of an HLA-A24restricted OY-TES-1 epitope recognized by cytotoxic T-cells.Microbiol Immunol.2005,49(11):1009-16.
11Tammela J,Uenaka A,Ono T,et al.OY-TES-1 expression and serum immunoreactivity in epithelial ovarian cancer.Int J Oncol.2006,29(4):903-10.
12范蓉,肖绍文,何少健等.精子蛋白Sp32/OY-TES.1基因mRNA表达的初步研究[J].解剖学报,2006,(37):102-10.
13王峰,范蓉,莫发荣等.癌睾丸抗原OY-TES-1基因的克隆、表达及纯化.中国免疫学杂志,2006,22:16-19
14Ono T,Sato S,Kimura N,et al.Serological analysis of BALB/C methylcholanthrene sarcoma Meth A by SEREX:identification of a cancer/testis antigen[J].Int J Cancer,2000,88(6):845-51
15Gure AO,Chua R,Williamson B,et al.Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer.Clin Cancer Res,2005,11(22):8055-8062
1.Ono T,Sa,to S,Kimura N,et al.Serological analysis of BALB/C methylcholanthrene sarcoma Meth A by SEREX:identification of a cancer / testis antigen[J].Int J Cancer,2000,88(6):845-851
2.Ono T,Kurashige T,Harada N,et al.Identification of proacrosin binding protein sp32 precursor as a human cancer/testis antigen[J].Proc Natl Acad Sci USA,2001,98(6):3282-3287
3.Scanlan MJ,Simpson AJ,Old LJ.The cancer/testis genes:Review,standardization,and commentary[J].Cancer Immunity,2004,4:1-15
4.Alsheimer M,Drewers T,Schutz W,et al.The cancer/testis antigen CAGE-1 is a component of the acrosome of spermatids and spermatozoa [J].Eur J Cell Biol,2005,84(223):445-452
5.Chen YT,Scanlan MJ,Sahin U,et al.Identification of cancer/testisantigengenes by massively parallel signature sequencing[J].Proc Natl Acad Sci USA,2005,102(22):7940-7945
6.Chen YT,Venditti CA,Theiler G,et al.Identification of CT46/HORM AD1,an immunogenic cancer/testis antigen encoding a putative meiosisrelatedprotein[J].Cancer Immunity,2005,5:9-17
7.Tureci O,Sahin U,Zwick C,et al.Identification of a meiosisspecific protein as a member of the class of cancer/testis antigens[J].Proc Natl Acad Sci USA,1998,95(9):5211-5216
8.王峰,范蓉,谢小薰.CT抗原-神经系统肿瘤免疫治疗的新靶向[J].肿 瘤防治研究,2007,34(2):154-156
9.Baba T,Niida Y,Michikawa Y,et al.An acrosomal protein,sp32,in mammalian sperm is a binding protein specific for two proacrosins and an acrosin intermediate[J].J Biol Chem,1994,269(13):10133-10140
10.Scanlan MJ,Simpson AJ,Old LJ.The cancer/testis genes:review,standardization,and commentary[J].Cancer Immun,2004,4:1
11.Kubuschok B,Xie X,Jesnowski R,et al.Expression of cancer testis antigens in pancreatic carcinoma cell lines,pancreatic adenocarcinoma and chronic pancreatitis[J].Int J Cancer,2004,109(4):568-575
12.范蓉,肖绍文,何少健,等.精子蛋白Sp32/OY-TES-1基因mRNA表达的探讨[J].解剖学报,2006,37(1):102-105
13.Okumura H,Noguchi Y,Uenaka A,et al.Identification of an HLA-A24-Restricted OY-TES-1 Epitope Recognized by Cytotoxic T-Cells[J].Microbiol Immunol.2005,49(11):1009-1016
14.Tammela J,Uenaka A,One T,et al.OY-TES-1 expression and serum immunoreactivity in epithelial ovarian cancer.Inter J Onco.2006,29:903-910
15.dos Santos NR,Torensma R,de Vries TJ,et al.Heterogeneous expression of the SSX cancer/testis antigens in human melanoma lesions and cell lines[J].Cancer Res.2000,60(6):1654-1662
2黄文清.神经肿瘤病理学(第二版).军事医学科学出版社,北京,2001,479-506.
3Sawamura Y,de Tribolet N.Immunobiology of brain tumor.Adv Tech Stand Neurosurg,1990,17:3-64.
4Pollack IF,Okada H,Chambers WH.Exploitation of immune mechanisms in the treatment of central nervous system cancer.Semin Pediatr Neurol,2000,7(2):131-143.
5Pallasch CP,Struss AK,Munnia A,et al.Autoantibodies against GLEA2and PHF3 in glioblastoma:tumor-associated autoantibodies correlated with prolonged survival.Int J Cancer.2005,117(3):456-9.
6Liu G,Ying H,Zeng G,et al.HER-2,gpl00,and MAGE-1 are expressed in human glioblastoma and recognized by cytotoxic T cells.Cancer Res,2004,64(14):4980-4986.
7Rodolfo M,Luksch R,Stockert E,et al.Antigen-specific immunity in neuroblastoma patients:antibody and T-cell recognition of NY-ESO-1tumor antigen.Cancer Res,2003,63(20):6948-6955.
8Chen YT,Scanlan MJ,Sahin U,et al.A testicular antigen aberrantly expressed in human cancers detected by autologous antibody screening.Proc Natl Acad Sci U S A,1997,94:1914.
9Ono T,Kurashige T,Harada N,et al.Identification of proacrosin binding protein sp32 precursor as a human cancer/testis antigen.Proc Natl Acad Sci U S A,2001,98:3282.
10Okumura H,Noguchi Y,Uenaka A,et al.Identification of an HLA-A24restricted OY-TES-1 epitope recognized by cytotoxic T-cells.Microbiol Immunol.2005,49(11):1009-16.
11Tammela J,Uenaka A,Ono T,et al.OY-TES-1 expression and serum immunoreactivity in epithelial ovarian cancer.Int J Oncol.2006,29(4):903-10.
12范蓉,肖绍文,何少健等.精子蛋白Sp32/OY-TES.1基因mRNA表达的初步研究[J].解剖学报,2006,(37):102-10.
13王峰,范蓉,莫发荣等.癌睾丸抗原OY-TES-1基因的克隆、表达及纯化.中国免疫学杂志,2006,22:16-19
14Ono T,Sato S,Kimura N,et al.Serological analysis of BALB/C methylcholanthrene sarcoma Meth A by SEREX:identification of a cancer/testis antigen[J].Int J Cancer,2000,88(6):845-51
15Gure AO,Chua R,Williamson B,et al.Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer.Clin Cancer Res,2005,11(22):8055-8062
1.Ono T,Sa,to S,Kimura N,et al.Serological analysis of BALB/C methylcholanthrene sarcoma Meth A by SEREX:identification of a cancer / testis antigen[J].Int J Cancer,2000,88(6):845-851
2.Ono T,Kurashige T,Harada N,et al.Identification of proacrosin binding protein sp32 precursor as a human cancer/testis antigen[J].Proc Natl Acad Sci USA,2001,98(6):3282-3287
3.Scanlan MJ,Simpson AJ,Old LJ.The cancer/testis genes:Review,standardization,and commentary[J].Cancer Immunity,2004,4:1-15
4.Alsheimer M,Drewers T,Schutz W,et al.The cancer/testis antigen CAGE-1 is a component of the acrosome of spermatids and spermatozoa [J].Eur J Cell Biol,2005,84(223):445-452
5.Chen YT,Scanlan MJ,Sahin U,et al.Identification of cancer/testisantigengenes by massively parallel signature sequencing[J].Proc Natl Acad Sci USA,2005,102(22):7940-7945
6.Chen YT,Venditti CA,Theiler G,et al.Identification of CT46/HORM AD1,an immunogenic cancer/testis antigen encoding a putative meiosisrelatedprotein[J].Cancer Immunity,2005,5:9-17
7.Tureci O,Sahin U,Zwick C,et al.Identification of a meiosisspecific protein as a member of the class of cancer/testis antigens[J].Proc Natl Acad Sci USA,1998,95(9):5211-5216
8.王峰,范蓉,谢小薰.CT抗原-神经系统肿瘤免疫治疗的新靶向[J].肿 瘤防治研究,2007,34(2):154-156
9.Baba T,Niida Y,Michikawa Y,et al.An acrosomal protein,sp32,in mammalian sperm is a binding protein specific for two proacrosins and an acrosin intermediate[J].J Biol Chem,1994,269(13):10133-10140
10.Scanlan MJ,Simpson AJ,Old LJ.The cancer/testis genes:review,standardization,and commentary[J].Cancer Immun,2004,4:1
11.Kubuschok B,Xie X,Jesnowski R,et al.Expression of cancer testis antigens in pancreatic carcinoma cell lines,pancreatic adenocarcinoma and chronic pancreatitis[J].Int J Cancer,2004,109(4):568-575
12.范蓉,肖绍文,何少健,等.精子蛋白Sp32/OY-TES-1基因mRNA表达的探讨[J].解剖学报,2006,37(1):102-105
13.Okumura H,Noguchi Y,Uenaka A,et al.Identification of an HLA-A24-Restricted OY-TES-1 Epitope Recognized by Cytotoxic T-Cells[J].Microbiol Immunol.2005,49(11):1009-1016
14.Tammela J,Uenaka A,One T,et al.OY-TES-1 expression and serum immunoreactivity in epithelial ovarian cancer.Inter J Onco.2006,29:903-910
15.dos Santos NR,Torensma R,de Vries TJ,et al.Heterogeneous expression of the SSX cancer/testis antigens in human melanoma lesions and cell lines[J].Cancer Res.2000,60(6):1654-1662