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

该项目旨在通过原位杂交（ISH），采用聚合酶链反应合成的高度生物素化探针，检测HTLV-1相关疾病，特别是恶性淋巴瘤（ML）细胞组成中的HTLV-1前病毒DNA及其信号。ISH未能检测到HTLV-1前病毒DNA，但在HTLV-1相关细胞系细胞和人ML细胞的石蜡切片中成功检测到其信号。HTLV-1前病毒DNA pX Tax区信号在包括成人T细胞白血病/淋巴瘤（ATLL）在内的T细胞多形性淋巴瘤（T-Pleo）中最常见。部分低度恶性T细胞淋巴细胞和其他淋巴细胞中的信号提示存在HTLV-1相关的除ATLL以外的其他淋巴细胞，以及HTLV-1相关低度恶性T细胞淋巴细胞向ATLL的促进作用。结果表明，HTLV-1 p40 tax的反式激活功能与HTLV-1前病毒DNA信号和宿主细胞基因激活的直接比较是不一致的。抗原修复法和ImmunoMax法可观察到ATLL细胞核内少量p40 tax蛋白。然后，考虑到HTLV-1 p40 tax的反式激活以外的功能，通过ISH和抗原修复以及ImmunoMax方法的组合，将能够分析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。

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.