Yu F, Itoyama Y, Kira J, Fujihara K, Kobayashi T, Kitamoto T, Suzumura A, Yamamoto N, Nakajima Y, Goto I
Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
J Immunol. 1994 Jun 15;152(12):5930-8.
Human T lymphotropic virus type I (HTLV-I) is linked to adult T cell leukemia as well as to HTLV-I-associated myelopathy/tropical spastic paraparesis. In this report, we studied the effects of HTLV-I-infected cell supernatants on HUVEC, fibroblasts, and glioma cells. The HTLV-I-infected cell supernatants (HUT102 and MT-2) strongly inhibited the proliferation of HUVEC, although they enhanced the proliferation of the fibroblasts. Regarding the glioma cells, only the MT-2 supernatant showed weak inhibitory effects on the proliferation. However, the HTLV-I-uninfected cell supernatants showed no effects on these target cells. The biologic activities of both HUT102 and MT-2 supernatants were found to be dose dependent and were reduced by heat treatment at 100 degrees C for 5 min, but not at 56 degrees C for 30 min. These activities were not dependent on the concentrations of HTLV-I viral particles and were only minimally affected by the presence of anti-HTLV-I Abs. A bioassay of various cytokines revealed that the activity of TNF was much higher in the HUT102 and MT-2 supernatants than in the HTLV-I-uninfected cell supernatants (MOLT-4, Jurkat, and K-562). rTNF-alpha and rTNF-beta also showed strong inhibitory effects on HUVEC as well as on the enhancement of the fibroblast growth. With the use of Sephadex G-100 column chromatography, we obtained the highest activities from the 60- through 70-kDa fractions of the HUT102 supernatant and some activities from the 20- through 30-kDa fractions. The biologic activities of both the whole HUT102 supernatant and its active fractions were completely blocked by anti-TNF-beta mAb, although they were not blocked by anti-TNF-alpha mAb. In a Western blot assay, the 25- and 27-kDa bands of TNF-beta were shown clearly in the HUT102 supernatant, although no TNF-alpha bands appeared. These findings suggest that TNF-beta is present in either its oligomeric or monomeric form in the HTLV-I-infected cell supernatants and is also mainly responsible for the supernatants' effects on HUVECs and fibroblasts.
人类嗜T淋巴细胞病毒I型（HTLV-I）与成人T细胞白血病以及HTLV-I相关脊髓病/热带痉挛性截瘫有关。在本报告中，我们研究了HTLV-I感染细胞的上清液对人脐静脉内皮细胞（HUVEC）、成纤维细胞和胶质瘤细胞的影响。HTLV-I感染细胞的上清液（HUT102和MT-2）强烈抑制HUVEC的增殖，尽管它们促进了成纤维细胞的增殖。对于胶质瘤细胞，只有MT-2上清液对其增殖显示出微弱的抑制作用。然而，未感染HTLV-I的细胞上清液对这些靶细胞没有影响。发现HUT102和MT-2上清液的生物活性均呈剂量依赖性，并且在100℃加热5分钟后活性降低，但在56℃加热30分钟后活性未降低。这些活性不依赖于HTLV-I病毒颗粒的浓度，并且仅受到抗HTLV-I抗体存在的最小影响。对各种细胞因子的生物测定显示，HUT102和MT-2上清液中肿瘤坏死因子（TNF）的活性远高于未感染HTLV-I的细胞上清液（MOLT-4、Jurkat和K-562）。重组TNF-α和重组TNF-β对HUVEC以及成纤维细胞生长的促进作用也显示出强烈的抑制作用。通过使用Sephadex G-100柱色谱法，我们从HUT102上清液的60至70 kDa组分中获得了最高活性，并从20至30 kDa组分中获得了一些活性。整个HUT102上清液及其活性组分的生物活性均被抗TNF-β单克隆抗体完全阻断，尽管它们未被抗TNF-α单克隆抗体阻断。在蛋白质印迹分析中，HUT102上清液中清晰显示出25 kDa和27 kDa的TNF-β条带，尽管未出现TNF-α条带。这些发现表明，TNF-β以其寡聚体或单体形式存在于HTLV-I感染细胞的上清液中，并且也是上清液对HUVEC和成纤维细胞产生作用的主要原因。
