Abstract：
BACKGROUND: Antibiotic-Refractory Lyme Arthritis (ARLA) involves a complex interplay of T cell responses targeting Borrelia burgdorferi antigens succeeding towards autoantigens by epitope spreading. However, the precise molecular mechanisms driving the pathogenic T cell response in ARLA remain unclear. Our aim was to elucidate the molecular program of disease-specific Th cells.
METHODS: Using flow cytometry, high-throughput T cell receptor (TCR) sequencing and scRNA-seq of CD4+ Th cells isolated from the joints of European ARLA patients, we aimed at inferring antigen specificity through unbiased analysis of TCR repertoire patterns, identifying surrogate markers for disease-specific TCRs and connecting TCR specificity to transcriptional patterns.
RESULTS: PD-1hiHLA-DR+CD4+ effector T cells were clonally expanded within the inflamed joints and persisted throughout disease course. Among these cells, we identified a distinct TCRβ motif restricted to HLA-DRB1*11 or *13 alleles. These alleles, being underrepresented in North American ARLA patients, were unexpectedly prevalent in our European cohort. The identified TCRβ motif served as surrogate marker for a convergent TCR response specific to ARLA, distinguishing it from other rheumatic diseases. In the scRNA-seq dataset, the TCRβ motif particularly mapped to peripheral T helper (TPH) cells displaying signs of sustained proliferation, continuous TCR signaling, and expressing CXCL13 and IFN-γ.
CONCLUSIONS: By inferring disease-specific TCRs from synovial T cells we identified a convergent TCR response in the joints of ARLA patients that continuously fueled the expansion of TPH cells expressing a pathogenic cytokine effector program. The identified TCRs will aid in uncovering the major antigen targets of the maladaptive immune response.
BACKGROUND: Supported by the German Research Foundation (DFG) MO 2160/4-1; the Federal Ministry of Education and Research (BMBF; Advanced Clinician Scientist-Program INTERACT; 01EO2108) embedded in the Interdisciplinary Center for Clinical Research (IZKF) of the University Hospital Würzburg; the German Center for Infection Research (DZIF; Clinical Leave Program; TI07.001_007) and the Interdisciplinary Center for Clinical Research (IZKF) Würzburg (Clinician Scientist Program, Z-2/CSP-30).
METHODS: Using flow cytometry, high-throughput T cell receptor (TCR) sequencing and scRNA-seq of CD4+ Th cells isolated from the joints of European ARLA patients, we aimed at inferring antigen specificity through unbiased analysis of TCR repertoire patterns, identifying surrogate markers for disease-specific TCRs and connecting TCR specificity to transcriptional patterns.
RESULTS: PD-1hiHLA-DR+CD4+ effector T cells were clonally expanded within the inflamed joints and persisted throughout disease course. Among these cells, we identified a distinct TCRβ motif restricted to HLA-DRB1*11 or *13 alleles. These alleles, being underrepresented in North American ARLA patients, were unexpectedly prevalent in our European cohort. The identified TCRβ motif served as surrogate marker for a convergent TCR response specific to ARLA, distinguishing it from other rheumatic diseases. In the scRNA-seq dataset, the TCRβ motif particularly mapped to peripheral T helper (TPH) cells displaying signs of sustained proliferation, continuous TCR signaling, and expressing CXCL13 and IFN-γ.
CONCLUSIONS: By inferring disease-specific TCRs from synovial T cells we identified a convergent TCR response in the joints of ARLA patients that continuously fueled the expansion of TPH cells expressing a pathogenic cytokine effector program. The identified TCRs will aid in uncovering the major antigen targets of the maladaptive immune response.
BACKGROUND: Supported by the German Research Foundation (DFG) MO 2160/4-1; the Federal Ministry of Education and Research (BMBF; Advanced Clinician Scientist-Program INTERACT; 01EO2108) embedded in the Interdisciplinary Center for Clinical Research (IZKF) of the University Hospital Würzburg; the German Center for Infection Research (DZIF; Clinical Leave Program; TI07.001_007) and the Interdisciplinary Center for Clinical Research (IZKF) Würzburg (Clinician Scientist Program, Z-2/CSP-30).
摘要：
背景：抗生素-难治性莱姆关节炎（ARLA）涉及T细胞应答的复杂相互作用，这些T细胞应答靶向伯氏疏螺旋体抗原，通过表位扩散成功转化为自身抗原。然而,驱动ARLA致病性T细胞应答的确切分子机制尚不清楚.我们的目的是阐明疾病特异性Th细胞的分子程序。
方法：使用流式细胞术，从欧洲ARLA患者关节中分离出的CD4+Th细胞的高通量T细胞受体（TCR）测序和scRNA-seq，我们旨在通过对TCR库模式的无偏分析来推断抗原特异性，识别疾病特异性TCR的替代标记，并将TCR特异性与转录模式联系起来。
结果：PD-1hiHLA-DR+CD4+效应T细胞在发炎的关节内克隆扩增，并在整个病程中持续存在。在这些细胞中,我们确定了一个独特的TCRβ基序，仅限于HLA-DRB1*11或*13等位基因。这些等位基因,在北美ARLA患者中代表性不足，在我们的欧洲队列中出乎意料地普遍存在。鉴定的TCRβ基序作为ARLA特异性趋同TCR反应的替代标记，区别于其他风湿性疾病。在scRNA-seq数据集中，TCRβ基序特别定位到显示持续增殖迹象的外周T辅助T（TPH）细胞，连续TCR信号，并表达CXCL13和IFN-γ。
结论：通过推断来自滑膜T细胞的疾病特异性TCR，我们鉴定了ARLA患者关节中的趋同TCR反应，该反应持续促进了表达致病性细胞因子效应程序的TPH细胞的扩增。鉴定的TCR将有助于揭示适应不良免疫应答的主要抗原靶标。
背景：由德国研究基金会（DFG）MO2160/4-1支持；联邦教育与研究部（BMBF；高级临床科学家计划INTERACT；01EO2108）嵌入维尔茨堡大学医院的跨学科临床研究中心（IZKF）；德国临床感染研究中心（DZIFüTIZ00临床研究计划）；Z-2/CSP-30)。
方法：使用流式细胞术，从欧洲ARLA患者关节中分离出的CD4+Th细胞的高通量T细胞受体（TCR）测序和scRNA-seq，我们旨在通过对TCR库模式的无偏分析来推断抗原特异性，识别疾病特异性TCR的替代标记，并将TCR特异性与转录模式联系起来。
结果：PD-1hiHLA-DR+CD4+效应T细胞在发炎的关节内克隆扩增，并在整个病程中持续存在。在这些细胞中,我们确定了一个独特的TCRβ基序，仅限于HLA-DRB1*11或*13等位基因。这些等位基因,在北美ARLA患者中代表性不足，在我们的欧洲队列中出乎意料地普遍存在。鉴定的TCRβ基序作为ARLA特异性趋同TCR反应的替代标记，区别于其他风湿性疾病。在scRNA-seq数据集中，TCRβ基序特别定位到显示持续增殖迹象的外周T辅助T（TPH）细胞，连续TCR信号，并表达CXCL13和IFN-γ。
结论：通过推断来自滑膜T细胞的疾病特异性TCR，我们鉴定了ARLA患者关节中的趋同TCR反应，该反应持续促进了表达致病性细胞因子效应程序的TPH细胞的扩增。鉴定的TCR将有助于揭示适应不良免疫应答的主要抗原靶标。
背景：由德国研究基金会（DFG）MO2160/4-1支持；联邦教育与研究部（BMBF；高级临床科学家计划INTERACT；01EO2108）嵌入维尔茨堡大学医院的跨学科临床研究中心（IZKF）；德国临床感染研究中心（DZIFüTIZ00临床研究计划）；Z-2/CSP-30)。
