The difficulty of detecting HTLV-1 proviral sequences in patients with mycosis fungoides.
The difficulty of detecting HTLV-1 proviral sequences in patients with mycosis fungoides.
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检测蕈样肉芽肿患者 HTLV-1 前病毒序列的困难。
DOI：
10.1097/00042560-199612010-00003
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Zucker-Franklin,D
中科院分区：
文献类型：
--
作者：
Pancake,BA;Zucker-Franklin,D
尽管大多数皮肤T细胞淋巴瘤患者，包括蕈样真菌病（MF）及其白血病变体ssamzary综合征，对人类T细胞淋巴病毒（HTLV- i /II）抗体血清呈阴性，但最近有研究表明，95%以上的此类患者携带与HTLV基因组编码转调控/转化基因tax区域相关的前病毒DNA序列。然而，HTLV序列的证明，即使经过聚合酶链反应（PCR）扩增，也没有普遍成功，一些研究者继续质疑这一观察结果。为了解决这一争议，我们将已发表的不太成功的方法与本实验室目前使用的技术进行了比较。主要差异在于：(a)所使用细胞的性质[新鲜分离的与培养的外周血单个核细胞（PBMC）]和(b)制备PCR样品的方法（全细胞裂解物与DNA提取物）。对10例不同MF患者的PBMC与其中1例患者的健康女儿的PBMC进行比较分析。虽然所有的PBMC裂解物都是阳性的，但这些个体中只有一个的DNA提取物显示了HTLV税基序列。研究还比较了来自7名不同MF患者的税务序列阳性PBMC培养细胞的细胞裂解物和DNA提取物。当全细胞裂解物被用作PCR扩增和Southern分析的靶物时，七个细胞中的四个细胞在不同的培养时间后显示保留了大量序列，而没有一个DNA提取物是阳性的。似乎使用全细胞裂解物代替DNA提取物，使用新鲜细胞代替培养细胞，大大提高了检测MF患者标本中HTLV-I税层序列的能力。
Although most patients with cutaneous T cell lymphomas, including mycosis fungoides (MF) and its leukemic variant, the Sézary syndrome, are seronegative for antibodies to the human T cell lymphotropic viruses (HTLV-I/II), it has recently been shown that> 95% of such patients harbor proviral DNA sequences related to the region of the HTLV genome that encodes the transregulatory/transforming gene, tax. However, the demonstration of HTLV sequences, even after amplification by polymerase chain reaction (PCR), has not been universally successful, and some investigators continue to question this observation. In an effort to resolve this controversy, we have compared published methodologies that have been less successful with techniques currently used in this laboratory. Major differences were found in (a) the nature of the cells used [freshly isolated versus cultured peripheral blood mononuclear cells (PBMC)] and (b) the methods used to prepare samples for PCR (whole cell lysates versus DNA extracts). PBMC from 10 different MF patients and the healthy daughter of 1 of the patients were subjected to comparative analyses. While all of the PBMC lysates were positive, the DNA extract from only one of these individuals revealed HTLV tax sequences. Studies were also conducted comparing cell lysates and DNA extracts of cultured cells derived from tax sequence-positive PBMC from seven different MF patients. The cells from four of the seven were shown to have retained tax sequences after varying times in culture, when whole-cell lysates were used as targets for PCR amplification and Southern analysis, whereas none of the DNA extracts were positive. It appears that the use of whole-cell lysates instead of DNA extracts and the use of fresh instead of cultured cells greatly enhance the ability to detect HTLV-I tax sequences in specimens from MF patients.
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影响因子：
6
作者：
I. Anagnostopoulos;M. Hummel;P. Kaudewitz;Hermann Herbst;O. Braun;H. Stein
通讯作者：
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影响因子：
11.4
作者：
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影响因子：
5.4
作者：
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通讯作者：
G. Khoury
HIV 感染者的皮肤疾病类似于蕈样肉芽肿，其皮肤和血细胞也含有前病毒 HTLV I 型。
影响因子：
1.5
作者：
Zucker-Franklin,D;Pancake,BA;Friedman-Kien,AE
通讯作者：
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删除蕈样肉芽肿患者血液和皮肤病变中的 HTLV-I 原病毒。
影响因子：
56.9
作者：
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通讯作者：
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