Kubota R, Kawanishi T, Matsubara H, Manns A, Jacobson S
Viral Immunology Section, Neuroimmunology Branch, National Institute of Neurological Disorders and Stroke, Bethesda, MD 20892, USA.
J Neuroimmunol. 2000 Jan 24;102(2):208-15. doi: 10.1016/s0165-5728(99)00175-7.
Human T lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is an inflammatory neurological disease caused by HTLV-I infection. It has been shown that HAM/TSP patients have high proviral loads and an extraordinarily high frequency of circulating CD8 + cytotoxic T lymphocytes specific for HTLV-I in their peripheral blood when compared to asymptomatic HTLV-I carriers (AC). We have previously described an intracellular cytokine detection assay, in which interferon-gamma (IFN-gamma) + CD8 + lymphocytes are specific for HTLV-I in infected individuals. Here, we have established a competitive polymerase chain reaction assay to measure the proviral load of patients and investigate a potential relationship between proviral load and virus-specific CD8 + lymphocytes. Genomic DNA was extracted from peripheral blood lymphocytes (PBL) from eight HAM/TSP patients and seven AC for the measurement of HTLV-I measuring proviral loads. The same PBL were analyzed for intracellular IFN-gamma expression by flow cytometry. In the HAM/TSP patients and AC, the average proviral loads were 34,482 and 9784 copy/microg DNA (P = 0.021), and the average of IFN-gamma + CD8 + lymphocytes in total PBL were 1.47 and 0.08% (P = 0.001), respectively. It was confirmed that HAM/TSP patients have both high proviral loads and increased HTLV-I-specific CD8 + lymphocytes. Furthermore, we found a positive correlation between both factors in the patients with HAM/TSP (P = 0.044) but not in the AC (P = 0.508). These findings suggest that the high number of HTLV-I-specific lymphocytes may result from the increased proviral load in HAM/TSP patients.
人类嗜T淋巴细胞病毒I型（HTLV-I）相关脊髓病/热带痉挛性截瘫（HAM/TSP）是一种由HTLV-I感染引起的炎性神经疾病。研究表明，与无症状HTLV-I携带者（AC）相比，HAM/TSP患者的前病毒载量较高，且外周血中循环的HTLV-I特异性CD8 + 细胞毒性T淋巴细胞频率极高。我们之前描述过一种细胞内细胞因子检测方法，其中干扰素-γ（IFN-γ）+ CD8 + 淋巴细胞在受感染个体中对HTLV-I具有特异性。在此，我们建立了一种竞争性聚合酶链反应检测方法来测量患者的前病毒载量，并研究前病毒载量与病毒特异性CD8 + 淋巴细胞之间的潜在关系。从8例HAM/TSP患者和7例AC的外周血淋巴细胞（PBL）中提取基因组DNA，用于测量HTLV-I的前病毒载量。通过流式细胞术分析相同的PBL中细胞内IFN-γ的表达。在HAM/TSP患者和AC中，平均前病毒载量分别为34,482和9784拷贝/μg DNA（P = 0.021），总PBL中IFN-γ + CD8 + 淋巴细胞的平均值分别为1.47%和0.08%（P = 0.001）。证实HAM/TSP患者既有高前病毒载量，又有HTLV-I特异性CD8 + 淋巴细胞增加。此外，我们发现HAM/TSP患者中这两个因素之间存在正相关（P = 0.044），而在AC中则无相关性（P = 0.508）。这些发现表明，HAM/TSP患者中HTLV-I特异性淋巴细胞数量增多可能是前病毒载量增加所致。
