In-situ-hybridization analysis of HTLV-1-related diseases, employing highly biotinylated probes synthesized by polymerase chain reaction

采用聚合酶链式反应合成的高度生物素化探针对 HTLV-1 相关疾病进行原位杂交分析

基本信息

批准号：
05670176
负责人：
HASUI Kazuhisa
金额：
$ 1.41万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

This project aimed to detect HTLV-1 proviral DNA and its signals in cellular composition of HTLV-1-related diseases, especially malignant lymphomas (MLs), by means of in-situ-hybridization (ISH) employing highly biotinylated probes synthesized by polymerase chain reaction. The ISH failed to detect HTLV-1 proviral DNA but succeeded to detect its signals in paraffin sections of HTLV-1-related cell line cells and human ML cells. The signals of HTLV-1 proviral DNA pX Tax region were recognized the most frequently in T-cell pleomorphic lymphoma (T-Pleo) including adult T-cell leukemia/lymphoma (ATLL). The signals in some of low-grade T-cell MLs and the other MLs suggested the existence of HTLV-1-related MLs other than ATLL and the promotion from HTLV-1-related low-grade T-cell MLs to ATLL.Antigen-retrieval immunohistochemical detection of HTLV-1 p40tax was performed in the MLs revealing the signals in ISH.Bur p40tax distributed in cytoplasm of ML cells in many T-cell MLs and in nuclei in a small number of T-cell MLs, suggesting that the direct comparision of the signals of HTLV-1 proviral DNA and the activation of host cell genes would fail to see the transactivating function of HTLV-1 p40tax. A small amount of p40tax in the nuclei of ATLL cells could be visualized by antigen-retrieval and ImmunoMax method. Then, the relation of the activation of HTLV-1 proviral DNA and host cell gens in HTLV-1-related diseases will be able tp be analyzed by means of the combination of the ISH and the antigen-retrieval and ImmunoMax method, considering the HTLV-1 p40tax's functions other than the transactivation.

该项目旨在通过采用聚合酶链合成的高度生物素化的探针进行原位杂交（ISH）来检测HTLV-1相关疾病，特别是恶性淋巴瘤（ML）的细胞成分中的HTLV-1前病毒DNA及其信号反应。 ISH 未能检测到 HTLV-1 前病毒 DNA，但成功地在 HTLV-1 相关细胞系细胞和人 ML 细胞的石蜡切片中检测到其信号。 HTLV-1 前病毒 DNA pX Tax 区的信号在 T 细胞多形性淋巴瘤 (T-Pleo) 包括成人 T 细胞白血病/淋巴瘤 (ATLL) 中最常见。一些低级别 T 细胞 ML 和其他 ML 中的信号表明存在除 ATLL 之外的 HTLV-1 相关 ML，以及从 HTLV-1 相关低级别 T 细胞 ML 向 ATLL 的促进。在 ML 中进行 HTLV-1 p40tax 的检索免疫组织化学检测，揭示 ISH 中的信号。Bur p40tax 分布在许多 ML 细胞的细胞质中。 T 细胞 ML 和少量 T 细胞 ML 的细胞核中，表明直接比较 HTLV-1 前病毒 DNA 的信号和宿主细胞基因的激活将无法看到 HTLV-1 p40tax 的反式激活功能。通过抗原修复和ImmunoMax方法可以观察到ATLL细胞核中的少量p40tax。然后，考虑HTLV，通过ISH与抗原修复和ImmunoMax方法相结合，分析HTLV-1相关疾病中HTLV-1前病毒DNA的激活与宿主细胞基因的关系-1 p40tax 的反式激活以外的功能。