MOLECULAR STUDIES OF ERYTHEMA MULTIFORME

描述：(改编自申请者的摘要)-多形性红斑(EM) 是由包括疱疹在内的多种诱因引起的过敏性反应。 单纯疱疹病毒(HSV)感染。从血液中鉴定出HSV DNA序列 病变，但未分离到传染性病毒。T细胞反应也是 与血吸虫病的发病机制有关，但他们的HSV特异性和功能 仍不清楚。申请者表明，病毒DNA清除在 红斑与单纯疱疹病毒病变相关，但临床红斑症状与单纯疱疹病毒相关 基因表达。单纯疱疹病毒DNA聚合酶基因(POL)在血液中表达 病变，它们的CD4+V-β2 T细胞和干扰素-γ呈阳性。痊愈了 皮损皮肤均为阴性。调查员假设哈姆是一名 由T细胞反应组合引起的免疫病理疾病 对HSV抗原(可能是POL)和病毒介导的病理效应。这个 具体目标是：(I)研究病毒(Pol)特异性T细胞的作用 从血液中建立T细胞系/克隆在血液病发病机制中的反应 皮损和愈合的皮肤并检测其HSV蛋白特异性和 属性(即干扰素-γ的产生)。将采用酶联免疫吸附试验斑点试验 估计皮肤中有反应的T细胞的克隆大小。两个T细胞克隆 和来自血吸虫病和单纯疱疹病毒患者的PBMC将被检查是否存在 皮肤归巢受体CLA。第二个目标(二)是研究POL在 通过测定POL辅助蛋白UL42的表达引起的表皮损伤 和POL催化活性，以及(B)诱导/激活 转染Pol或Pol/UL42的角质形成细胞中转化生长因子-β和p21waf的表达 与生长停滞/细胞凋亡有关。目的(III)是确定单纯疱疹病毒DNA的路径 输送到皮肤上。这将通过确定HSV DNA的存在来完成 血液中CLA+(和CD34+/CLA+)朗格汉斯细胞(LC)前体的片段 和单纯疱疹病毒患者，并检测这些细胞分解单纯疱疹病毒的能力 DNA/Pol DNA在体外成熟为LC时保留。这些研究将提供 更好地了解血肿的发病机制及其与单纯疱疹病毒的关系， 从而允许基于靶向的新疗法的开发 疾病相关单纯疱疹病毒基因(S)。它们还将产生与临床相关的 分化型血液病的诊断和治疗信息 从其他病原体或药物敏感引起的EM患者中分离出来。

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.