Development of preditection method to prevent occurrence of adult T-cell leukemia/lymphoma (ATLL)

本项目进行了以下调查，旨在预测成人t细胞白血病/淋巴瘤（ATLL）的发生。1、制备包括ATLL、HTLV-1携带者及其他淋巴细胞增生的白血病外周血组织标本（PBTS）。2、建立了新的简化催化信号扩增（nsCSA）系统——超敏免疫组化。3、采用抗htlv -1 Tax抗体的nsCSA系统显示，包括PBTS在内的组织标本中ATLL细胞中有极少量的Tax表达。4、作为htlv -1感染细胞的肿瘤特征，通过nsCSA系统检测Ki67抗原和p53蛋白的表达。(1) HTLV-1携带者的粒细胞分节核被抗ki67抗原抗体标记，提示HTLV-1感染对造血有影响。(2) p53 1801抗体的nsCSA系统检测到p53与凋亡相关的表达，因为p53中调节其凋亡功能的Ser46被p53 1801和…More抗体标记。(3)在ATLL、HTLV-1携带者和传染性单核细胞增多症患者中，p53磷酸化的Ser392被p53Phos nsCSA系统标记，提示p53失活。(4) HTLV-1携带者中p53Phos和p53 1801抗体的表达在老年人群中显著（Kruskal-Walli试验）。5、引入经典甘氨酸处理，阻断一抗非特异性反应，建立自身免疫染色的多重免疫染色方法。6、利用网络上的DNA数据库，利用寡核苷酸探针检测目标DNA的mRNA，可以确定DNA序列。已知杂交溶液和检测系统是mRNA原位杂交的重要因素。利用玻片表面温度传感器，可以实现原位PCR法玻片上组织切片温度的控制。下一阶段检测系统预计引入nsCSA系统（291字）

This project performed following investigations, aiming to predict occurrence of adult T-cell leukemia/lymphoma (ATLL). 1,Peripheral blood tissue specimens (PBTS) of leukemia including ATLL, HTLV-1 carriers and the other lymphocytosis were prepared. 2,Supersensitive immunohistochemistry, new simplified catalyzed signal amplification (nsCSA) system, was established. 3,By the nsCSA system employing anti-HTLV-1 Tax antibody, it was shown that an extremely small amount of Tax was expressed in ATLL cells in tissue specimens includings PBTS. 4,As neoplastic features of HTLV-1-infected cells, expression of Ki67 antigen and p53 protein was examined by the nsCSA system. (1)In HTLV-1 carriers, segmented nuclei of granulocytes were labeled by anti-Ki67 antigen antibody, suggesting an effect of HTLV-1 infection on hematopoiesis. (2)The nsCSA system of p53 1801 antibody detected expression of p53 related to apoptosis, since Ser46 in p53 regulating its function in apoptosis is labeled by p53 1801 an … More tibody. (3)Phosphorylated Ser392 in p53 was labeled by the nsCSA system of p53Phos in ATLL, HTLV-1 carriers and infectious mononucleosis, suggesting inactivation of p53. (4)Expression of p53Phos and p53 1801 antibody was significantly observed in advam\ncing age of HTLV-1 carriers (Kruskal-Walli test). 5,Multi-immunostaing method by an autoimmunostainer was established by introducing classic glycine treatment and blocking non-specific reaction of primary antibody. 6, By the DNA database on the web, it was possible to determine DNA sequence of oligonucleotide probe detecting mRNA of the target DNA. It was known that hybridization solution and detection system to visualize hybridized oligonucleotide probes were important factors in mRNA in-situ hybridization. By means of a sensor of slide glass surface temperature, it was possible to control temperature of tissue section on the slide glass in in-situ PCR method. The next stage detection system expects introduction of the nsCSA system (291 words) Less