Poiesz B J, Ruscetti F W, Gazdar A F, Bunn P A, Minna J D, Gallo R C
Proc Natl Acad Sci U S A. 1980 Dec;77(12):7415-9. doi: 10.1073/pnas.77.12.7415.
Retrovirus particles with type C morphology were found in two T-cell lymphoblastoid cell lines, HUT 102 and CTCL-3, and in fresh peripheral blood lymphocytes obtained from a patient with a cutaneous T-cell lymphoma (mycosis fungoides). The cell lines continuously produce these viruses, which are collectively referred to as HTLV, strain CR(HTLV(CR)). Originally, the production of virus from HUT 102 cells required induction with 5-iodo-2'-deoxyuridine, but the cell line became a constitutive producer of virus at its 56th passage. Cell line CTCL-3 has been a constitutive producer of virus from its second passage in culture. Both mature and immature extracellular virus particles were seen in thin-section electron micrographs of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle has been seen. Mature particles were 100-110 nm in diameter, consisted of an electron-dense core surrounded by an outer membrane separated by an electron-lucent region, banded at a density of 1.16 g/ml on a continuous 25-65% sucrose gradient, and contained 70S RNA and a DNA polymerase activity typical of viral reverse transcriptase (RT; RNA-dependent DNA nucleotidyltransferase). Under certain conditions of assay, HTLV(CR) RT showed cation preference for Mg(2+) over Mn(2+), distinct from the characteristics of cellular DNA polymerases purified from human lymphocytes and the RT from most type C viruses. Antibodies to cellular DNA polymerase gamma and anti-bodies against RT purified from several animal retroviruses failed to detectably interact with HTLV(CR) RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLV(CR) RT to cellular DNA polymerases gamma or RT of these viruses. Six major proteins, with sizes of approximately 10,000, 13,000, 19,000, 24,000, 42,000, and 52,000 daltons, were apparent when doubly banded, disrupted HTLV(CR) particles were chromatographed on a NaDodSO(4)/polyacrylamide gel. The number of these particle-associated proteins is consistent with the expected proteins of a retrovirus, but the sizes of some are distinct from those of most known retroviruses of the primate subgroups.
在两种T细胞淋巴母细胞系HUT 102和CTCL - 3以及一名皮肤T细胞淋巴瘤（蕈样肉芽肿）患者的新鲜外周血淋巴细胞中发现了具有C型形态的逆转录病毒颗粒。这些细胞系持续产生这些病毒，它们统称为HTLV，CR株（HTLV(CR)）。最初，从HUT 102细胞产生病毒需要用5 - 碘 - 2'-脱氧尿苷诱导，但该细胞系在第56代时成为病毒的组成型生产者。细胞系CTCL - 3从其在培养中的第二代起就是病毒的组成型生产者。在固定、沉淀的细胞材料的超薄切片电子显微照片中可见成熟和未成熟的细胞外病毒颗粒；偶尔还可见典型的C型出芽病毒颗粒。未观察到任何形式的细胞内病毒颗粒。成熟颗粒直径为100 - 110纳米，由一个电子致密核心组成，周围是一个由电子透明区域分隔的外膜，在连续的25 - 65%蔗糖梯度上以1.16克/毫升的密度进行区带分离，并含有70S RNA和典型的病毒逆转录酶（RT；RNA依赖性DNA核苷酸转移酶）的DNA聚合酶活性。在某些测定条件下，HTLV(CR) RT对Mg(2+)的阳离子偏好高于Mn(2+)，这与从人淋巴细胞中纯化的细胞DNA聚合酶以及大多数C型病毒的RT的特性不同。在各自同源DNA聚合酶呈阳性的条件下，针对细胞DNA聚合酶γ 的抗体和从几种动物逆转录病毒中纯化的抗RT抗体未能与HTLV(CR) RT发生可检测到的相互作用，这表明HTLV(CR) RT与细胞DNA聚合酶γ 或这些病毒的RT缺乏密切关系。当将双重区带、破坏的HTLV(CR)颗粒在NaDodSO(4)/聚丙烯酰胺凝胶上进行色谱分析时，出现了六种主要蛋白质，大小约为10,000、13,000、19,000、24,000、42,000和52,000道尔顿。这些与颗粒相关的蛋白质数量与逆转录病毒预期的蛋白质一致，但其中一些的大小与灵长类亚组中大多数已知逆转录病毒的不同。
