DETECTION AND ISOLATION OF TYPE-C RETROVIRUS PARTICLES FROM FRESH AND CULTURED LYMPHOCYTES OF A PATIENT WITH CUTANEOUS T-CELL LYMPHOMA
DETECTION AND ISOLATION OF TYPE-C RETROVIRUS PARTICLES FROM FRESH AND CULTURED LYMPHOCYTES OF A PATIENT WITH CUTANEOUS T-CELL LYMPHOMA
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DOI：
10.1073/pnas.77.12.7415
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
GALLO, RC
中科院分区：
文献类型：
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作者：
POIESZ, BJ;RUSCETTI, FW;GALLO, RC
在两种T细胞淋巴母细胞系HUT 102和CTCL - 3以及从一名皮肤T细胞淋巴瘤（蕈样肉芽肿）患者获取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，它们被统称为人类皮肤T细胞淋巴瘤病毒CR株（HTLVCR）。最初，从HUT 102细胞中产生病毒需要用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性产生者。细胞系CTCL - 3从培养的第2代起就一直是病毒的组成性产生者。在固定的、沉淀的细胞物质的超薄切片电子显微镜下可以看到成熟和未成熟的细胞外病毒颗粒；有时还能看到典型的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，核心外有一层外膜，两者之间由一个电子透明区域隔开，在连续的25 - 65%蔗糖梯度中以1.16克/毫升的密度成带，并且含有70S RNA和一种具有病毒逆转录酶（RT；依赖RNA的DNA核苷酸转移酶）典型特征的DNA聚合酶活性。在某些测定条件下，HTLVCR RT表现出对Mg²⁺比对Mn²⁺的阳离子偏好，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。针对细胞DNA聚合酶γ的抗体以及针对从几种动物逆转录病毒中纯化的RT的抗体，在各自同源DNA聚合酶呈阳性的条件下，未能与HTLVCR RT发生可检测的相互作用，这表明HTLVCR RT与细胞DNA聚合酶γ或这些病毒的RT没有密切关系。当对经过两次成带、裂解的HTLVCR颗粒在十二烷基硫酸钠/聚丙烯酰胺凝胶上进行层析时，明显出现六种主要蛋白质，其大小约为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数量一致，但其中一些蛋白质的大小与灵长类亚群中大多数已知逆转录病毒的不同。
Retrovirus particles with type C morphology were found in 2 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 human cutaneous T-cell lymphoma virus strain CR (HTLVCR). 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 2nd passage in culture. Both mature and immature extracellular virus particles were seen in thin-section EM of fixed, pelleted cellular material; on occasion, typical type C budding virus particles were seen. No form of intracellular virus particle was 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, HTLVCR RT showed cation preference for Mg2+ over Mn2+, 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 antibodies against RT purified from several animal retroviruses failed to detectably interact with HTLVCR RT under conditions that were positive for the respective homologous DNA polymerase, demonstrating a lack of close relationship of HTLVCR 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 HTLVCR particles were chromatographed on a sodium dodecyl sulfate/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.