MOLECULAR STUDIES OF ERYTHEMA MULTIFORME

描述：（改编自申请人摘要）-多形性红斑（EM） 是一种由包括疱疹在内的许多刺激剂引起的超敏反应 单纯疱疹病毒感染（HAEM）。在HAEM中鉴定HSV DNA序列 病变，但未分离出感染性病毒。T细胞反应也是 与HAEM发病机制有关，但它们的HSV特异性和功能 目前仍不清楚.申请人表明，病毒DNA清除受损， HAEM与HSV病变相关，但临床HAEM症状与HSV相关 基因表达。HSV DNA聚合酶基因（Pol）在HAEM中表达， 病变，并且它们对CD 4 + V-β-2 T细胞和IFN-γ呈阳性。愈合 皮损均为阴性。研究者假设HAEM是一种 由T细胞应答的组合引起的免疫病理学疾病 HSV抗原（可能是Pol）和病毒介导的病理效应。的 具体目的是：（I）研究病毒（Pol）特异性T细胞的作用 通过从HAEM建立T细胞系/克隆在HAEM发病机制中的应答 损伤和愈合的皮肤，并确定它们的HSV蛋白特异性， 特性（即IFN-γ产生）。将使用ELISA斑点试验来 估计皮肤中应答T细胞的克隆大小。两个T细胞克隆 将检查来自HAEM和HSV患者的PBMC中存在的 皮肤归巢受体CLA。第二个目的（II）是研究Pol在 通过测定（a）Pol辅助蛋白UL 42的表达 和Pol催化活性，和（B）诱导/激活 用Pol或Pol/UL 42转染的角质形成细胞中的TGF-β和p21 waf， 与生长停滞/凋亡有关。目的（III）明确HSV DNA的传播途径 运输到皮肤。这将通过确定HSV DNA的存在来完成 来自HAEM的CLA+（和CD 34 +/CLA+）朗格汉斯细胞（LC）前体中的片段 以及检测这些细胞裂解HSV的能力 DNA/保留Pol DNA，因为它们在体外成熟为LC。这些研究将提供 更好地了解HAEM发病机制及其与HSV的关系， 从而允许开发基于靶向的新型疗法 疾病相关HSV基因。它们还将产生临床相关的 诊断和管理HAEM患者的信息， 由其他病原体或药物敏感性引起的EM患者。

DESCRIPTION: (Adapted from the applicant's abstract) - Erythema multiforme (EM) is a hypersensitivity reaction caused by many provoking agents including herpes simplex virus (HSV) infection (HAEM). HSV DNA sequences were identified in HAEM lesions, but infectious virus was not isolated. T cell responses were also implicated in HAEM pathogenesis, but their HSV specificity and function are still unclear. The applicants showed that viral DNA clearance is impaired in HAEM relative to HSV lesions, but clinical HAEM symptoms correlate with HSV gene expression. The HSV DNA polymerase gene (Pol) is expressed in HAEM lesions, and they are positive for CD4+ V-beta-2 T cells and IFN-gamma. Healed lesional skin were all negative. The investigator hypothesizes that HAEM is an immunopathological disease resulting from the combination of T cell responses to HSV antigens (likely Pol) and virus-mediated pathological effects. The specific aims are: (I) To examine the role of virus (Pol)-specific T cell responses in HAEM pathogenesis by establishing T cell lines/clones from HAEM lesional and healed skin and determining their HSV protein specificity and properties (viz. IFN-gamma production). ELISA spot assay will be used to estimate the clonal size of responding T cells in the skin. Both T cell clones and PBMC from HAEM and HSV patients will be examined for the presence of the skin homing receptor CLA. The second aim (II) is to examine the role of Pol in epidermal damage by determining(a) expression of the Pol accessory protein UL42 and Pol catalytic activity in HAEM lesions, and (b) the induction/activation of TGF-beta and p21waf in keratinocytes transfected with Pol or Pol/UL42 as it relates to growth arrest/apoptosis. Aim (III) is to define the route of HSV DNA transport to the skin. This will be done by determining the presence of HSV DNA fragments in CLA+ (and CD34+/CLA+) Langerhans cells (LC) precursors from HAEM and HSV patients and examining the ability of these cells to fragment HSV DNA/retain Pol DNA as they mature into LC in vitro. These studies will provide a better understanding of HAEM pathogenesis and its relationship to HSV, thereby allowing the development of novel therapies based on the targeting of disease-related HSV gene(s). They will also generate clinically relevant information for the diagnosis and management of HAEM patients as differentiated from those with EM caused by other pathogens or by drug sensitivity.