BORIS和CT45两种新的CT抗原单克隆抗体的制备及其蛋白水平表达和分布的初步研究

发布时间：2018-01-03 03:38

本文关键词：BORIS和CT45两种新的CT抗原单克隆抗体的制备及其蛋白水平表达和分布的初步研究　出处：《第四军医大学》2007年硕士论文　论文类型：学位论文

【摘要】： 随着肿瘤免疫学的发展,越来越多的肿瘤相关抗原被应用于肿瘤的免疫研究中。肿瘤/睾丸抗原(Cancer/Testis Antigens, CTA)具有可局限性表达于肿瘤组织并且可以在肿瘤患者体内引起免疫反应等特点,因而成为近年来肿瘤免疫研究的热点。但是目前CT抗原的研究大多停留在DNA及mRNA的水平上,蛋白水平上的研究还相对滞后,主要原因是缺乏特异性强敏感性高的单克隆抗体。 BORIS和CT45是两种新近发现的CT抗原,为了进一步在蛋白水平上对其进行研究,我们首次制备BORIS及CT45的单克隆抗体,并对其在蛋白水平上的表达和分布进行了初步的研究。应用淋巴细胞杂交瘤技术,制备和鉴定了BORIS和CT45两种CT抗原的单克隆抗体,并筛选出特异性强及敏感性高能识别天然抗原的相应单克隆抗体:FMU-BORIS13和CT45-03,免疫组织化学染色最佳工作浓度分别为4.5μg/ml和2.6μg/ml。以获得的单抗为工具,运用免疫组织化学染色的方法在睾丸组织正常乳腺组织乳腺良性疾病组织乳腺癌及肺癌组织上对两种CT抗原在蛋白水平上的分布进行了初步的研究。结果发现在睾丸组织中:两种CT抗原主要分布在生精细胞发育的早期即主要表达在精原细胞及初级精母细胞的细胞核中。在乳腺癌组织中BORIS呈高水平的表达,表达率为85.5% (94/110);在正常乳腺组织及乳腺良性疾病组织中也检测到了BORIS的表达,但表达水平远低于乳腺癌组织;在肺癌组织中CT45表达百分率较低仅为4.8%(3/62)。两种CT抗原在肿瘤组织中的表达与肿瘤组织的分期分级无明显相关性。应用单克隆抗体在睾丸组织及肿瘤组织对两种抗原进行初步的检测,证实了两种CT抗原在蛋白水平上的分布符合CT抗原的特点。两种新的CT抗原的单克隆抗体的获得,为蛋白水平上进一步研究两种CT抗原提供了有力的工具。
[Abstract]:With the development of tumor immunology, immune of tumor associated antigen was applied in more and more tumors. The cancer / testis antigens (Cancer/Testis, Antigens, CTA) has the limitations expressed in tumor tissue and can cause immune reactions etc. in cancer patients, so it has become a hot research topic in recent years. But the tumor immunity at present study on CT antigen mostly remained at DNA and mRNA level, the research on the protein level is still lagging behind, the main reason is the lack of specificity of monoclonal antibody with high sensitivity.?
BORIS and CT45 are two newly discovered CT antigens. In order to further research on protein level, we prepared BORIS and CT45 monoclonal antibodies for the first time, and studied their expression and distribution at protein level.
The application of hybridoma technology and preparation of monoclonal antibody BORIS and CT45 two CT antigen were identified and screened the corresponding monoclonal antibody specificity and sensitivity of high recognition of natural antigens: FMU-BORIS13 and CT45-03, immunohistochemical staining of optimal concentration were 4.5 g/ml and 2.6 g/ml. to obtain the monoclonal antibody for the tool, using immunohistochemical staining method in testicular tissue of normal breast tissue?? breast benign disease? Breast cancer and lung cancer tissue of two CT antigen distribution at protein level were studied. The results showed that in testis: two CT antigen mainly distributed in the early development of spermatogenic cell that is mainly expressed in spermatogonia and primary spermatocyte nuclei. In breast cancer the expression of BORIS was high level, the expression rate was 85.5% (94/110) in normal breast group; Fabric and benign breast disease tissues also detected the expression of BORIS, but the expression level is far lower than that of breast cancer; in lung cancer tissues positive percentage of CT45 was only 4.8% (3/62). The expression of two CT antigen in tumor tissues and tumor staging had no obvious correlation.
Application of monoclonal antibody to detection of these two antigens in testicular tissue and tumor tissue, confirmed the distribution of two kinds of CT antigen at the protein level in accordance with the characteristics of CT antigen. Monoclonal antibodies of two kinds of new CT antigen, which provides a powerful tool for further research on the protein level of two CT antigen.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

【共引文献】