HTLV-I MYCOSIS FUNGOIDES/SEZARY SYNDROME

真菌病(MF)及其白血病变种Sezary综合征(SS)， 是最常见的皮肤T细胞淋巴瘤。流行病学 研究表明，MF/SS的发病率正在增加，在 绝对值和占所有淋巴瘤的比例。统计数字也 表明HTLV-I型逆转录病毒感染的发病率在#年上升。 美国逆转录为前病毒DNA后，HTLV-I 整合到宿主T细胞的基因组中。潜伏期过后 20-30岁，另一种形式的皮肤T细胞淋巴瘤被称为成人T- 细胞白血病/淋巴瘤(ATL)在1%的感染者中发生。ATL 据报道，美国有病例，特别是在东南部和 夏威夷。尽管MF/SS患者具有典型的血清阴性的特点 HTLV-I使用常规检测，最新超微结构，分子 生物学和血清学结果表明，这种逆转录病毒或其变种 可能在MF/SS的发病中起一定作用。我们未公布的数据支持这一点 查看。我们筛选了38-58名来自不同地区的MF患者的活检组织。 用聚合酶链式反应检测HTLV-I前病毒DNA的地理区域 HTLV-I的Px、Pol和env基因，并用 Southern杂交分析。其中11例为HTLV-I+，包括6例 PX+Polenv病例、两例PX+Polo+env病例、两例pXpol+env病例和一例 PXpol+env案例。11例HTLV-I+病例包括4/5的加利福尼亚州病例和 1/1智利案例。相比之下，只有6/29的俄亥俄地区病例，0/15的瑞士病例， 0/5的马萨诸塞州病例，没有其他地理分布的病例 均为HTLV-I+。此外，来自俄亥俄州的10-17名非MF/SS皮肤病控制人员 相应的HTLV-I聚合酶链式反应产物均为阴性。聚合酶链式反应产物现在已经 从MT4阳性对照细胞系中克隆并测序，来自 PX+Pol+MF患者和1例env+MF患者。与 来自日本的HTLV-I世界流行株显示相同的PX和POL 序列，但独特的环境序列。这些结果是最广泛的 到目前为止，它们的种类是确凿的。Y提供了非常有说服力 支持HTLV-I或相关逆转录病毒参与的假设 在至少一些MF/SS的发病机制中，并提示了一种潜在的 地理集群。这一假设得到了我们的进一步支持 免疫印迹法检测MF/SS患者血清的高灵敏度分析 用重组的HTLV蛋白。总体而言，23名MF/SS患者中有10人表现出 HTLV血清反应性的一些证据，包括7/13环太平洋病例 相比之下，俄亥俄州只有1/8的案件。我们在这项研究提案中的目标是 通过使用PCR/Southern杂交分析来验证和扩展我们的发现 获取更多数量和地理多样性的HTLV-I基因 在MF/SS患者中。我们还将使用DNA克隆和测序技术 确定不同HTLV-I=之间是否有任何共同点 MF/SS病例，无论是前病毒的基因结构还是前病毒的位置 整合。除了有助于阐明MF/SS的发病机制外， 这为诊断、预防和治疗提供了新的方法 研究可能对理解发病机制有更广泛的影响。 与MF/SS相关的其他类型的淋巴瘤。

Mycosis fungoides (MF) and its leukemic variant, the Sezary syndrome (SS), are the most common forms of cutaneous T-cell lymphoma. Epidemiological studies indicate that the incidence of MF/SS is increasing, both in absolute terms and as a proportion of all lymphomas. Statistics also indicate that the incidence of HTLV-I retrovirus infection is increasing in the U.S.A. Following reverse transcription into proviral DNA, HTLV-I becomes integrated into the genome of host T-cells. After a latent period of 20-30 years, another form of cutaneous T-cell lymphoma known as adult T- cell leukemia/lymphoma (ATL) develops in 1% of infected individuals. ATL cases have been reported in the U.S.A., especially in the Southeast and Hawaii. Although MF/SS patients are characteristically seronegative for HTLV-I using conventional assays, recent ultrastructural, molecular biologic and serologic findings suggest that this retrovirus or a variant may play a role in MF/SS pathogenesis. Our unpublished data support this view. We have screened biopsies from 38-58 MF patients from different geographical regions for HTLV-I proviral DNA using a PCR-based assay for the HTLV-I pX, pol and env genes and have confirmed our findings using Southern blot analysis. Eleven of these cases were HTLV-I+ including six pX+polenv cases, two pX+pol+env cases, two pXpol+env cases and one pXpol+env case. The eleven HTLV-I+ cases included 4/5 California cases and 1/1 Chile case. In contrast only 6/29 Ohio area cases, 0/15 Swiss cases, 0/5 Massachusetts cases and none of other geographically scattered cases were HTLV-I+. Furthermore, 10-17 non-MF/SS skin disease controls from Ohio were negative for corresponding HTLV-I PCR products. PCR products have now been cloned and sequenced from the mT4 positive control cell line, from a pX+pol+ MF patient and from an env+ MF patient. Comparison to a cosmopolitan strain of HTLV-I from Japan indicates identical pX and pol sequences but unique env sequences. These results are the most extensive and definitive of their kind to date. The y provide very compelling support for the hypothesis that HTLV-I or a related retrovirus is involved in the pathogenesis of at least some cases of MF/SS and suggest a potential geographical clustering. This hypothesis is supported further by our highly sensitive analysis of MF/SS patient sera using Western blots spiked with recombinant HTLV proteins. Overall, 10/23 MF/SS patients have shown some evidence of HTLV seroreactivity, including 7/13 pacific Rim cases versus only 1/8 Ohio cases. Our objective in this research proposal is to confirm and extend our findings by using PCR/Southern blot analysis to test for additional HTLV-I genes in a larger number and geographical diversity of MF/SS patients. We will also use DNA cloning and sequencing techniques to determine if there are any common denominators among different HTLV-I= MF/SS cases, either in terms of proviral gene structure or site of proviral integration. In addition to helping clarify the pathogenesis of MF/SS and providing new approaches to diagnosis, prevention and therapy, this research could have broader implications for understanding the pathogenesis of other types of lymphomas that are associated with MF/SS.