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

本项目旨在利用聚合酶链式反应合成的高度生物素化的探针进行原位杂交(ISH)，检测HTLV-1相关疾病，特别是恶性淋巴瘤(MLS)细胞成分中的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T细胞MLs和其他MLs中的信号提示，除ATLL外，还存在HTLV-1相关的MLs，并从HTLV-1相关低度T细胞MLs向ATLL的促进作用。抗原恢复免疫组织化学检测显示，HTLV-1p40Tax在MLS中有信号，但分布在许多T细胞MLL的ML细胞胞浆和少量T细胞MLL的细胞核中，提示HTLV-1前病毒DNA信号与宿主细胞基因激活的直接比较不能看到HTLV-1 p40Tax的反式激活功能。抗原修复和免疫MAX法可见少量的p40Tax分布于ATLL细胞的胞核。然后，考虑到HTLV-1 p40Tax的反式激活以外的功能，采用ISH、抗原修复和免疫MAX相结合的方法，分析HTLV-1相关疾病中HTLV-1前病毒DNA的激活与宿主细胞抗原的关系。

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.