HTLV-I MYCOSIS FUNGOIDES/SEZARY SYNDROME

蕈样肉芽肿（MF）及其白血病变体，塞扎里综合征（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， 绝对值和占所有淋巴瘤的比例。 统计还 表明HTLV-I逆转录病毒感染的发病率在增加， 在逆转录成前病毒DNA后，HTLV-I 整合到宿主T细胞的基因组中。 经过一段潜伏期 20-30岁，另一种形式的皮肤T细胞淋巴瘤称为成人T细胞淋巴瘤， 细胞白血病/淋巴瘤（ATL）在1%的感染个体中发展。 ATL 在美国也有病例报告，尤其是在东南部， 夏威夷。 虽然MF/SS患者的特征性血清阴性， HTLV-I使用常规测定，最近的超微结构，分子 生物学和血清学结果表明，这种逆转录病毒或其变体 可能在MF/SS发病机制中起作用。 我们未公布的数据支持这一点 风景 我们筛选了38-58例MF患者的活检， HTLV-I前病毒DNA的地理区域，使用基于PCR的测定， HTLV-I pX，pol和env基因，并证实了我们的发现， Southern印迹分析。 其中11例为HTLV-I+，包括6例 pX+ pol + env病例2例，pX+pol+env病例2例，pX + pol +env病例1例 pXpol+env case. 11例HTLV-I+病例包括4/5例加州病例， 1/1智利病例。 相比之下，只有6/29的俄亥俄州地区病例，0/15的瑞士病例， 0/5例马萨诸塞州病例，无其他地理分散病例 HTLV-I+。 此外，来自俄亥俄州的10-17名非MF/SS皮肤病对照 相应的HTLV-Ⅰ PCR产物均为阴性。 PCR产品目前 从mT4阳性对照细胞系中克隆并测序， pX+pol+ MF患者和env+ MF患者。 相比于 来自日本的HTLV-I的世界性菌株显示相同的pX和pol 序列，但独特的env序列。 这些结果是最广泛的 也是迄今为止最具权威性的 y提供了非常引人注目的 支持HTLV-I或相关逆转录病毒参与的假设 在至少一些MF/SS病例的发病机制中， 地理集群。 这一假设得到了我们的进一步支持。 使用加标蛋白质印迹法对MF/SS患者血清进行高灵敏度分析 重组HTLV蛋白。 总体而言，10/23例MF/SS患者显示 HTLV血清反应性的一些证据，包括7/13例环太平洋病例 而俄亥俄州只有1/8的病例。 我们在这项研究计划中的目标是 通过PCR/Southern印迹分析证实并扩展了我们的发现， 对于更多数量和地理多样性的额外HTLV-I基因， MF/SS患者 我们还将使用DNA克隆和测序技术 以确定不同HTLV-I之间是否存在任何共同的干扰因素， MF/SS病例，无论是在前病毒基因结构或前病毒位点方面， 一体化 除了有助于阐明MF/SS的发病机制外， 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意义， 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 Our unpublished data support this view.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.