Role of Chemokines in Innate and Adaptive Immunity in the Lung

项目摘要 慢性铍病（CBD）是由工作场所的铍（Be）暴露引起的，其特征是： 肉芽肿性炎症和Be特异性，Th 1型分泌亮氨酸的CD 4 + T细胞的积累， 肺Be暴露还与细胞因子/趋化因子分泌的诱导和协调的细胞因子/趋化因子分泌有关。 先天性和其他适应性细胞向肺的募集。我们的初步数据显示，暴露于HLA-DP 2 Tg小鼠对氧化铍（BeO）的耐受性以及对源自CBD的支气管肺泡灌洗（BAL）细胞的刺激 BeSO_4组CCL_3和CCL_4分泌均显著增加。重要的是，我们已经确定了相关的 来源于CCL 3和CCL 4的肽，其被表达的TCR识别为Be依赖性新抗原 对来自表达HLA-DP 2的CBD患者的BAL的CD 4 + T细胞的作用。Be负载的HLA-DP 2-CCL 3和 HLA-DP 2-CCL 4四聚体染色显示这些配体被10-40%的IFN-γ表达的细胞识别， BAL中的Be特异性CD 4 + T细胞，支持这些T细胞在Be诱导的免疫应答中的重要性。 反应总的来说，我们的数据表明，先天介质，CCL 3和CCL 4，分泌到肺 同时也作为被大部分Be特异性T细胞识别的关键T细胞表位被靶向。 CD 4 + T细胞。因此，在持续性抗原暴露的情况下，我们假设Be诱导的CCL 3和CCL 4表达增加。 CCL 4分泌产生了先天性和适应性免疫激活的破坏性循环， 表位特异性CD 4 + T细胞与基因工程化T细胞将调节Be诱导的肺部炎症。 使用HLA-DP 2 Tg小鼠，Aim 1将确定BeO诱导的CCL 3和CCL 4的细胞来源和作用 分泌作为肺部炎症的先天介质，而Aim 2将跟踪CCL 3/Be-和 CBD鼠模型中的CCL 4/Be特异性CD 4 + T细胞。最终目标是利用嵌合抗原受体 (CAR)在表达HLA-DP 2的小鼠和CBD患者中靶向表位特异性CD 4 + T细胞的T细胞， 潜在的治疗方法在治疗铍诱导的疾病。拟议研究的意义 依赖于新的趋化因子的鉴定，这些趋化因子不仅参与先天性炎性细胞的募集， 编码Be修饰的自身肽，其结合HLA-DP 2并代表CBD中的显性T细胞表位。的 成功完成这些研究，使用创新技术消除HLA中的表位特异性T细胞， 表达DP 2的小鼠和患有Be介导的疾病的人将导致CAR T细胞作为免疫调节剂的潜在用途。 CBD是一种全身性皮质类固醇替代品很少的疾病。

Project Summary Chronic beryllium disease (CBD) results from beryllium (Be) exposure in the workplace and is characterized by granulomatous inflammation and the accumulation of Be-specific, Th1-type cytokine-secreting CD4+ T cells in the lung. Be exposure is also associated with the induction of cytokine/chemokine secretion and the coordinated recruitment of innate and other adaptive cells to the lung. Our preliminary data show that exposure of HLA-DP2 Tg mice to Be oxide (BeO) as well as stimulation of bronchoalveolar lavage (BAL) cells derived from CBD patients with BeSO4 both induced significant CCL3 and CCL4 secretion. Importantly, we have identified related peptides derived from CCL3 and CCL4 that are recognized as Be-dependent neoantigens by TCRs expressed on CD4+ T cells derived from the BAL of HLA-DP2-expressing CBD patients. Be-loaded HLA-DP2-CCL3 and HLA-DP2-CCL4 tetramer staining showed that these ligands were recognized by 10-40% of IFN-γ-expressing, Be-specific CD4+ T cells in the BAL, supporting the importance of these T cells in the Be-induced immune response. Collectively, our data demonstrate that innate mediators, CCL3 and CCL4, are secreted into the lung in response to Be while also being targeted as key T cell epitopes recognized by a large fraction of Be-specific CD4+ T cells. Thus, in the setting of persistent antigen exposure, we hypothesize that Be-induced CCL3 and CCL4 secretion creates a destructive cycle of innate and adaptive immune activation and that depletion of these epitope-specific CD4+ T cells with genetically-engineered T cells will modulate Be-induced lung inflammation. Using HLA-DP2 Tg mice, Aim 1 will determine the cellular source and role of BeO-induced CCL3 and CCL4 secretion as innate mediators of lung inflammation while Aim 2 will track the dynamics of CCL3/Be- and CCL4/Be-specific CD4+ T cells in a murine model of CBD. The final aim will utilize chimeric antigen receptor (CAR) T cells to target epitope-specific CD4+ T cells in HLA-DP2-expressing mice and CBD patients as a potential therapeutic approach in the treatment of Be-induced disease. The significance of the proposed study rests on the novel identification of chemokines involved not only in innate inflammatory cell recruitment, but also encoding Be-modified self-peptides that bind to HLA-DP2 and represent dominant T cell epitopes in CBD. The successful completion of these studies using innovative technology to deplete epitope-specific T cells in HLA- DP2-expressing mice and humans with Be-mediated disease will lead to the potential use of CAR T cells as a therapeutic option for CBD, a disease with few alternatives to systemic corticosteroids.