MOLECULAR PATHOLOGICAL ANALYSIS OF NERVOUS DISEASES CAUSED BY CHRONIC VIRAL INFECTION

为了阐明慢性病毒感染所致神经系统疾病的发病机制，我们首先采用HTLV-I前病毒原位聚合酶链式反应和细胞表面标志物免疫组织化学双重染色的方法，研究了HTLV-I感染细胞在HAM/TSP脊髓损伤中的定位。HTLV-I前病毒DNA定位于浸润性UCHL-1+T细胞，尤其是血管周围。HTLV-I+细胞数与病情活动度呈正相关，与病程呈负相关。采用TaqMan定量聚合酶链式反应方法检测202例HAM/TSP患者和206例HTLV-I携带者外周血单核细胞前病毒载量。HAM/TSP患者的前病毒载量明显高于健康携带者，且与疾病进展率、抗HTLV-I抗体效价、抗-HTLV-I抗体滴度、抗HTLV-I抗体滴度呈正相关。在HAM/TSP pa…的外周血单核细胞和脑脊液细胞中也发现了高前病毒载量和频繁的Tax蛋白表达更多采用原位聚合酶链式反应和高灵敏免疫组织化学技术。这些结果提示HTLV-I前病毒载量及其抗原的表达在HAM/TSP的致病机制中具有重要作用，是一种降低病毒载量的治疗方法，其表达可能适用于HAM/TSP的治疗。我们还探讨了HTLV-I携带者持续感染的一种模式。在纵向收集的500份血液标本中，所有HTLV-I血清阳性样本的HTLV-I前病毒的PCR检测均为阳性，而所有HTLV-I血清阴性的血液样本均为HTLV-I前病毒阴性。这表明，血清阴性的HTLV-I携带者是罕见的，在HTLV-I感染中，来自血清阴性献血者的感染可能并不常见。共收集了100例扁桃体切除患者的扁桃体，并检查了HTLV-I感染细胞的定位。所有HTLV-I阳性患者的扁桃体边缘带面积增大，仅在这些区域检测到HTLV-I前病毒。这些发现表明，在HTLV-I感染者中，扁桃体可能是持续感染的部位和感染细胞的储存库。较少

In order to clarify pathogenesis of the nervous diseases caused by chronic viral infection, we firstly investigated localization of HTLV-I infected cells in the spinal cord lesion of HAM/TSP using double staining of in situ PCR for HTLV-I provirus and immunohistochemistry of cell surface markers. HTLV-I proviral DNA was localized at infiltrated UCHL-1+ T-cells especially in the perivascular areas. Numbers of HTLV-I+ cells were positively correlated with the disease activity and negatively with duration of illness. In addition, proviral loads of peripheral blood mononuclear cells in 202 HAM/TSP patients and 206 HTLV-I carriers were measured by quantitative PCR using TaqMan PCR method. The proviral load of HAM/TSP patients was much higher than that of healthy carrier and correlated with the disease progression rate, anti-HTLV-I Ab titers, and the values of. neopterin in CSF.A high proviral load and frequent Tax protein expression were also demonstrated in PBMC and CSF cells of HAM/TSP pa … More tients using in situ PCR and highly sensitive immunohistochemistry technique. Taken together, these findings suggested that HTLV-I proviral load and expression of its antigens are important in pathogenic mechanism of HAM/TSP and a therapeutic method to reduce viral load and its expression may be suitable for the treatment of HAM/TSP.We also investigated a mode of persistent infection in HTLV-I carriers. Among 500 blood samples collected from a longitudinal series of patients, all HTLV-I sero-positive samples were positive for PCR detection of HTLV-I provirus and all of HTLV-I sero-negative blood samples were negative for HTLV-I provirus. This suggested that sero-negative HTLV-I carrier is rare and infection from a sero-negative blood donner may not be frequent in HTLV-I infection. Totally 100 tonsils were collected from a series of patients who received tonsillectomy and examined localization of HTLV-I infected cells. All tonsils from HTLV-I sero-positive patients showed increased area of marginal zone and HTLV-I provirus was detected only from such areas. These findings suggested a possibility that the tonsil might be a site of persistent infection and a reservoir of infected cells in HTLV-I infected individuals. Less