Borrelia burgdorferi-Induced Autoimmunity in Lyme Disease

这项资助旨在描述感染后莱姆病(LD)的自身免疫特征。 名为抗生素难治性莱姆关节炎(LA)的综合征，治疗后唯一的LD综合征 已经为其定义了特定的病理。我们之前曾报道过，过度 炎症，免疫失调的T细胞/Treg细胞比率，上调某些 MicroRNAs和感染诱导的自身免疫是这种不好结果的特征。 此外，难治性LA的最大遗传风险因素是某些HLA-DR等位基因，而我们 直接从这些患者的滑膜组织中鉴定出具有免疫原性的HLA-DR递呈多肽 病人。通过这种方式，我们已经证明了4种自身抗原，内皮细胞生长因子(ECGF)， 基质金属蛋白酶-10、载脂蛋白B-100和膜联蛋白A2是T和B细胞反应的靶点。 LD的每一种表现的患者；以及近一半使用抗生素的患者- 难治性LA对一种或多种自身抗原有自身抗体反应。基于RA- SEQ数据，我们在此报告难治性LA的滑膜病变具有高度的 炎性表达特征，包括与干扰素相关的基因上调- 反应，MHC II类抗原的处理和递呈，细胞介导的细胞毒性，以及 细胞增殖。我们现在提出滑膜成纤维细胞样滑膜细胞(FLS)，最 病变中常见的细胞，成为非常规的抗原提呈细胞(UAPC)，而CD4+ 具有细胞毒活性的T细胞可能针对FLS。正如目标1中详细说明的那样，我们有 在LA患者中发现了两种具有细胞毒性潜力的CD4+SLAMF7+T细胞，我们将 用单细胞RNA-seq法进一步确定其表型。在目标2中，我们将确定一个更大的 由专业APC或uAPC(FLS)呈递给CD4+T细胞的一系列HLA-DR-肽， 我们将描述CD4+SLAMF7+T细胞与细胞内FLS之间的分子相互作用 文化。在目标3中，我们提供了难治性LA中自身抗体可能 参与这一疾病过程。在这些患者中，高水平的IgG4自身抗体 ECGF、MMP-10和apoB-100分别与显著的纤维化和闭塞的微血管相关 滑膜组织的损伤。我们将评估结合特性和葡聚糖 这些自身抗体的组成从抗炎转变为促炎 难治性患者的表型。最后，我们将确定自身抗体的用途。 作为早期识别患者的诊断平台的一部分 适应不良免疫反应，这可能允许早期治疗来改善或防止这种情况 感染后综合症。

This grant seeks to characterize autoimmune features of a post-infectious Lyme disease (LD) syndrome called antibiotic-refractory Lyme arthritis (LA), the only post-treatment LD syndrome for which a specific pathology has been defined. We have previously reported that excessive inflammation, immune dysregulation of the Teff/Treg cell ratio, up-regulation of certain microRNAs, and infection-induced autoimmunity are features of this untoward outcome. Moreover, the greatest genetic risk factor for refractory LA is certain HLA-DR alleles, and we have identified immunogenic HLA-DR-presented peptides directly from synovial tissue in these patients. In this way, we have shown that 4 autoantigens, endothelial cell growth factor (ECGF), MMP-10, apoB-100, and annexin A2, are targets of T and B cell responses in subsets of patients with each of the manifestations of LD; and nearly half of patients with antibiotic- refractory LA have autoantibody responses to 1 or more of these autoantigens. Based on RA- seq data, we report in this grant that the synovial lesion in refractory LA has a highly inflammatory expression signature, which includes up-regulation of genes associated with IFN- -responses, MHC class II antigen processing and presentation, cell-mediated cytotoxicity, and cell proliferation. We now propose that synovial fibroblast-like synoviocytes (FLS), the most common cell in the lesion, become unconventional antigen presenting cells (uAPC), and CD4+ T cells with cytotoxic potential may be directed against FLS. As detailed in Aim 1, we have identified two types of CD4+ SLAMF7+ T cells with cytotoxic potential in LA patients, and we will further determine their phenotype using single-cell RNA-seq. In Aim 2, we will identify a greater range of HLA-DR-peptides presented to CD4+ T cells by professional APCs or uAPC (FLS), and we will delineate molecular interactions between CD4+SLAMF7+T cells and FLS in cell cultures. In Aim 3, we present preliminary data that autoantibodies in refractory LA may participate in this disease process. In these patients, high levels of IgG4 autoantibodies to ECGF, MMP-10 and apoB-100 each correlate with marked fibrosis and obliterative microvasular lesions in synovial tissue. We will assess whether the binding characteristics and glycan composition of these autoantibodies shift from an anti-inflammatory to a pro-inflammatory phenotype in refractory patients. Finally, we will determine the utility of autoantibody determinations as part of a diagnostic platform for early identification of patients with maladaptive immune responses, which may allow earlier therapy to ameliorate or prevent this post-infectious syndrome.