Specificity and Molecular Definition of Pathogenic Lymphocytes in Sjogren's Syndrome

干燥综合征致病性淋巴细胞的特异性和分子定义

• 批准号: 10469419
• 负责人: A Darise Farris
• 金额: $ 52.06万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-10 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10469419
• 关键词: Acinus organ component; Address; Amino Acid Receptors; Antibodies; Antibody Specificity; Antigen Presentation; Antigen Targeting; Antigenic Specificity; Antigens; Arthritis; Autoantigens; Autoimmune Diseases; Automobile Driving; B-Cell Lymphomas; B-Lymphocytes; Binding; Biological Response Modifier Therapy; Biopsy; CD30L; CD4 Positive T Lymphocytes; Cell Differentiation process; Cell Line; Cells; Characteristics; Chronic Disease; Clinical Markers; Clonal Expansion; Dental caries; Disease; Duct (organ) structure; Enzyme-Linked Immunosorbent Assay; Eye; Fatigue; Fibrosis; Frequencies; Gene Expression Profile; Genetic Transcription; Helper-Inducer T-Lymphocyte; Human; Immune response; Immune system; Immunoglobulin Class Switching; Immunologics; Immunoprecipitation; In Situ Hybridization; In Vitro; Individual; Label; Lacrimal gland structure; Lead; Learning; Link; Lymphocyte; Lymphocytic Infiltrate; MHC Class II Genes; Malignant - descriptor; Methods; Molecular; Monoclonal Antibodies; Mutate; Nervous system structure; Neurologic; Neurologic Symptoms; Neurotransmitter Receptor; Neurotransmitters; Oral; Oral Manifestations; Oral cavity; Pain; Pathogenicity; Pathology; Pathway interactions; Patients; Peptides; Phenotype; Plasmablast; Production; Property; Proteome; Research; Saliva; Salivary; Salivary Gland Diseases; Salivary Gland Tissue; Salivary Glands; Severities; Signal Transduction; Site; Sjogren' s Syndrome; Specificity; Structure of germinal center of lymph node; T-Cell Proliferation; T-Cell Receptor; T-Lymphocyte; TNFRSF8 gene; Testing; Tooth Loss; Transcript; Translating; Work; Xerostomia; antigen binding; autoimmune rheumatologic disease; autoreactivity; cell type; deep sequencing; design; diagnostic biomarker; effective therapy; eye dryness; genetic risk factor; immunoreaction; in vitro Assay; novel; novel diagnostics; oral infection; overexpression; patient subsets; peripheral blood; programs; retroviral transduction; single cell analysis; single-cell RNA sequencing; transcriptome

PROJECT SUMMARY Sjögren's syndrome (SS) is a rheumatic autoimmune disease selectively targeting salivary and lacrimal glands, leading to painful dry mouth and eyes, oral infections, severe dental caries/tooth loss, fatigue, arthritis, neurologic involvement, and malignant B cell lymphoma. There are no effective biologic therapies. The strongest genetic risk factor is MHC class II DR3/DQ2. CD4+ T cells dominate salivary gland (SG) focal lymphocytic infiltrates, but B cells are also present. CD4+ T cells drive germinal center formation and class-switched, somatically hyper- mutated antibodies, all characteristic features of SS SG. Identification of the antigens and immunologic pathways driving the aberrant immune responses in SS is the focus of our proposal. In previous work, we isolated paired T cell receptor (TCR) α and β sequences of clonally expanded SG CD4+ T cells from DR3/DQ2+ patients and showed that the degree of clonal expansion correlated with reduced salivary flow and increased SG fibrosis. These are the first findings to link SG T cell infiltrates with low salivary flow and SG damage. Expanded clones within and between patients showed TCR amino acid similarities at sites contacting antigenic peptide, providing evidence that common antigen(s) drive SG CD4+ T cell proliferation. We used SG plasmablast-derived monoclonal antibodies (mAbs) from the same patients to identify 15 new candidate autoantigens. We also identified key transcripts overexpressed in SG CD4+ T cells that are characteristic of T follicular helper (Tfh) cells or may promote Tfh differentiation (TNFSF8-encoded CD30L). This proposal will test three hypotheses: i) previously unidentified antigens are targeted by SG CD4+ T cells and plasmablasts in SS, ii) autoreactivity to novel antigens may explain oral and neurologic manifestations of SS, and iii) clonally expanded SG CD4+ T cells express a unique transcriptional signature that promotes Tfh cell differentiation. Aim 1. Determine whether shared TCRs expressed on clonally expanded SG CD4+ T cells are specific for canonical and novel SS antigens bound by SG plasmablasts of the same subjects and are present in peripheral blood (PB) of SS cases, with frequencies reflecting SG disease features and severity. Aim 2. Identify the specificities of SS salivary gland plasmablast-derived mAbs and determine whether they have functional consequences and/or cross react with canonical SS antigens. Aim 3. Determine the transcriptional program of SG CD4+ T cells expressing shared, disease-associated TCRs, validate their predicted phenotypes and their localization in SG tissue, and test the hypothesis that CD30/CD30L signaling promotes human Tfh differentiation. This high impact research will identify autoantigens and SG T cell properties that may explain oral and neurologic SS pathology and that may be exploited to eliminate or regulate lymphocytes that drive chronic disease.

项目总结 干燥综合征(SS)是一种选择性地以唾液和泪腺为靶点的风湿性自身免疫性疾病， 导致口腔和眼睛疼痛干燥，口腔感染，严重的龋齿/牙齿脱落，疲劳，关节炎，神经学 受累和恶性B细胞淋巴瘤。目前还没有有效的生物疗法。最强的基因 危险因素为MHC II类DR3/DQ2。唾液腺(SG)局部淋巴细胞浸润以CD4+T细胞为主， 但B细胞也存在。CD4+T细胞驱动生发中心的形成和类别转换，躯体高... 突变的抗体，都是SS SG的特征。抗原和免疫途径的鉴定 在SS中驱动异常的免疫反应是我们建议的重点。在之前的工作中，我们分离出了成对的 克隆扩增的DR3/DQ2+患者SGCD4+T细胞的T细胞受体α和β序列 克隆性扩张程度与唾液流量减少和SG纤维化增加相关。 这是首次发现SG T细胞浸润与低唾液流量和SG损伤有关。扩展克隆 患者内部和患者之间在与抗原肽接触的部位显示TCR氨基酸相似，提供 共同抗原(S)刺激SGCD4+T细胞增殖的证据。我们使用了SG浆母细胞衍生的 用来自同一患者的单抗(MAbs)识别15个新的候选自身抗原。我们也 已确定的关键转录本在SG CD4+T细胞中过度表达，这是T滤泡辅助者(TFH)的特征 细胞或可能促进TFH分化(TNFSF8编码的CD30L)。这项提议将检验三个假设：i) 在SS，II)自身反应性中，以前未知的抗原被SG的CD4+T细胞和浆母细胞靶向 新的抗原可以解释SS的口腔和神经学表现，以及III)克隆扩增的SG CD4+T细胞 表达独特的转录信号，促进TFH细胞分化。 目的1.确定克隆扩增的SG CD4+T细胞上表达的共享TCR是否是特异性的 规范的和新的SS抗原与相同受试者的SG浆母细胞结合并存在于 SS患者的外周血(PB)，其频率反映了SG的疾病特征和严重程度。 目的2.鉴定SS唾液腺成浆细胞来源的单抗的特异性，并确定 它们具有功能后果和/或与典型的SS抗原交叉反应。 目的3.确定SG CD4+T细胞的转录程序，表达共享的、疾病相关的 TCRs，验证其预测的表型及其在SG组织中的定位，并检验假设 CD30/CD30L信号通路促进人Tfh分化。 这项高影响力的研究将确定自身抗原和SG T细胞特性，可能解释口腔和神经学 SS病理，这可能被利用来消除或调节驱动慢性病的淋巴细胞。

项目成果

Genome-wide association study identifies Sjögren's risk loci with functional implications in immune and glandular cells.

• DOI: 10.1038/s41467-022-30773-y
• 发表时间: 2022-07-27
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Khatri, Bhuwan;Tessneer, Kandice L.;Rasmussen, Astrid;Aghakhanian, Farhang;Reksten, Tove Ragna;Adler, Adam;Alevizos, Ilias;Anaya, Juan-Manuel;Aqrawi, Lara A.;Baecklund, Eva;Brun, Johan G.;Bucher, Sara Magnusson;Eloranta, Maija-Leena;Engelke, Fiona;Forsblad-d'Elia, Helena;Glenn, Stuart B.;Hammenfors, Daniel;Imgenberg-Kreuz, Juliana;Jensen, Janicke Liaaen;Johnsen, Svein Joar Auglaend;Jonsson, Malin, V;Kvarnstrom, Marika;Kelly, Jennifer A.;Li, He;Mandl, Thomas;Martin, Javier;Nocturne, Gaetane;Norheim, Katrine Braekke;Palm, Oyvind;Skarstein, Kathrine;Stolarczyk, Anna M.;Taylor, Kimberly E.;Teruel, Maria;Theander, Elke;Venuturupalli, Swamy;Wallace, Daniel J.;Grundahl, Kiely M.;Hefner, Kimberly S.;Radfar, Lida;Lewis, David M.;Stone, Donald U.;Kaufman, C. Erick;Brennan, Michael T.;Guthridge, Joel M.;James, Judith A.;Scofield, R. Hal;Gaffney, Patrick M.;Criswell, Lindsey A.;Jonsson, Roland;Eriksson, Per;Bowman, Simon J.;Omdal, Roald;Ronnblom, Lars;Warner, Blake;Rischmueller, Maureen;Witte, Torsten;Farris, A. Darise;Mariette, Xavier;Alarcon-Riquelme, Marta E.;Shiboski, Caroline H.;Wahren-Herlenius, Marie;Ng, Wan-Fai;Sivils, Kathy L.;Adrianto, Indra;Nordmark, Gunnel;Lessard, Christopher J.
• 通讯作者: Lessard, Christopher J.

Dysregulated long non-coding RNA in Sjögren's disease impacts both interferon and adaptive immune responses.

• DOI: 10.1136/rmdopen-2022-002672
• 发表时间: 2022-11
• 期刊: RMD OPEN
• 影响因子: 6.2
• 作者: Joachims, Michelle L.;Khatri, Bhuwan;Li, Chuang;Tessneer, Kandice L.;Ice, John A.;Stolarczyk, Anna M.;Means, Nicolas;Grundahl, Kiely M.;Glenn, Stuart B.;Kelly, Jennifer A.;Lewis, David M.;Radfar, Lida;Stone, Donald U.;Guthridge, Joel M.;James, Judith A.;Scofield, R. Hal;Wiley, Graham B.;Wren, Jonathan D.;Gaffney, Patrick M.;Montgomery, Courtney G.;Sivils, Kathy L.;Rasmussen, Astrid;Farris, A. Darise;Adrianto, Indra;Lessard, Christopher J.
• 通讯作者: Lessard, Christopher J.