Detection of human T-lymphotropic virus type I p40tax protein in cerebrospinal fluid cells from patients with human T-lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis.
Detection of human T-lymphotropic virus type I p40tax protein in cerebrospinal fluid cells from patients with human T-lymphotropic virus type I-associated myelopathy/tropical spastic paraparesis.
复制标题
人类嗜T淋巴细胞病毒I型相关性脊髓病/热带痉挛性截瘫患者脑脊液细胞中人类嗜T淋巴细胞病毒I型p40tax蛋白的检测。
作者：
T. Moritoyo;S. Izumo;H. Moritoyo;Yuetsu Tanaka;Yoshimasa Kiyomatsu;M. Nagai;K. Usuku;M. Sorimachi;M. Osame
我们研究了人类T淋巴细胞病毒I型相关脊髓病(HAM)/热带痉挛麻痹(TSP)患者脑脊液(CSF)细胞和外周血单核细胞(PBMC)中人类T淋巴细胞病毒I型(HTLV-I)病毒转录本的作用。为了检测HTLV-I p40Tax蛋白，我们建立了一种结合酪胺信号扩增和定量分析的新的敏感的免疫组织化学方法。对7例HAM/TSP患者进行了检查。作为对照，四名患有其他神经系统疾病的患者接受了检查；其中两名患者感染了HTLV-I，另外两名患者没有感染。脑脊液细胞和外周血单个核细胞均与抗p40Tax蛋白的单抗LT-4反应，再用辣根过氧化物酶标记的二抗反应。用直接用酪胺标记的荧光素进行观察，并用激光扫描细胞仪对阳性细胞进行定量。在HAM/TSP患者的样本中，通过酪胺信号扩增成功地检测到HTLV-I p40Tax蛋白，但不是没有。HAM/TSP患者脑脊液细胞HTLV-Ip40Tax蛋白阳性率为0.04~1.16%，PBMCs阳性率为0.02~0.54%。HAM/TSP患者和HTLV-I携带者脑脊液细胞表达HTLV-Ip40Tax蛋白的频率均高于PBMC，且在病程较短的HAM/TSP患者中表达频率更高。这项技术可能成为研究HTLV-I相关疾病致病机制的有力工具。
We investigated the role of viral transcripts of human T-lymphotropic virus type I (HTLV-I) in the cerebrospinal fluid (CSF) cells and peripheral blood mononuclear cells (PBMCs) of patients with human T-lymphotropic virus type I-associated myelopathy (HAM)/tropical spastic paraparesis (TSP). To detect the HTLV-I p40tax protein, we developed a new sensitive method of immunohistochemistry combined with tyramide signal amplification and quantitative analysis. Seven patients with HAM/TSP were examined. As controls, four patients with other neurological diseases were examined; two of these patients were infected with HTLV-I and the other two were not. Both the CSF cells and PBMCs were reacted with a monoclonal antibody, Lt-4, for p40tax protein, followed by secondary antibody labeled with horseradish peroxidase. This was visualized by fluorescein directly labeled with tyramide and the number of positive cells was quantified with a Laser Scanning Cytometer. In the samples from patients with HAM/TSP, the HTLV-I p40tax protein was successfully detected by tyramide signal amplification, but not without it. In HAM/TSP patients, 0.04-1.16% of the CSF cells and 0.02-0.54% of PBMCs were positive for the HTLV-I p40tax protein, respectively. The expression of the HTLV-I p40tax protein in the CSF cells was more frequent than that in PBMCs in both HAM/TSP patients and HTLV-I carriers, and was also more frequent in the patients with HAM/TSP of shorter duration of illness. This technique could be a powerful tool to investigate the pathogenic mechanism of diseases associated with HTLV-I.