Chemokines Induce Wnt-Frizzled Gene Expression in Human T Cells

趋化因子诱导人类 T 细胞中 Wnt 卷曲基因表达

• 批准号: 7964049
• 负责人: DENNIS D. TAUB
• 金额: $ 29.01万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7964049
• 关键词: Adhesions; Adrenal Glands; Binding; Biology; Bone Marrow; CCL19 gene; CXCL12 gene; CXCR4 gene; Cells; Cellular biology; Chemotactic Factors; Chemotaxis; Data; Development; Event; Family member; GTP-Binding Proteins; Gene Expression; Gene Family; Glycoproteins; HIV; HIV Envelope Protein gp120; HIV-1; Human; Immigration; Immune; In Vitro; Inflammation; Inflammatory; Leukocytes; Ligands; Ligation; Liver; Lung; Maintenance; Mediating; Mediator of activation protein; Membrane Microdomains; Microarray Analysis; Modeling; Mus; Neoplasm Metastasis; Organogenesis; Pathway interactions; Play; Protein Family; Protein Kinase C; Receptor Activation; Receptor Signaling; Recombinants; Reporting; Rest; Rodent; Role; Signal Pathway; Signal Transduction; Stream; Structure; Surface; System; T-Lymphocyte; Thymoma; Thymus Gland; Time; Tissues; Virus; Wnt proteins; Work; blastomere structure; cell motility; cell type; chemokine; chemokine receptor; in vivo; lymph nodes; member; migration; novel; receptor; receptor expression; response; seven-transmembrane G-protein-coupled receptor; trafficking

Chemokines have been shown to induce and direct adhesion, chemotaxis, activation, and degranulation of human and rodent leukocytes both in vitro and in vivo. CXCL12 and CCL19 are two important chemokines that regulate T cell motility and activation under normal and inflammatory conditions. Despite numerous reports examining the function of chemokines, little is known about the transcriptional events involved therein. Here, we performed microarray analysis on CXCL12- treated T-cells, and found that the Wnt family of proteins was significantly upregulated during CXCL12 treatment. Confirmation of these results by real-time PCR and Western analysis revealed that the expression of Wnt5A and other members of the non-canonical Wnt pathway were specifically upregulated during CXCL12 stimulation, while -catenin and canonical Wnt family members were selectively downregulated. Wnt5A was found to augment signaling through the CXCL12-CXCR4 axis via the activation of protein kinase C (PKC). Moreover, our data has revealed that Wnt5A expression is required to mediate directional T-cell migration in response to CXCL12, and that the treatment of human T-cells with recombinant Wnt5A sensitized T-cells to CXCL12-induced migration. Furthermore,Wnt5A expression was also required for the sustained expression of CXCR4, both transcriptionally and translationally. These results were further supported in vivo using EL4 thymoma metastasis as a model of T-cell migration. Together, these data demonstrate, for the first time, that Wnt5A is a critical mediator in CXCL12-CXCR4 signaling and migration in human and murine T cells. Interestingly, we also found that Wnt10A plays a role in CCL19 chemotaxis and in the maintenance of CCR7 expression on T cells. These findings may reveal a novel cooperative signaling network between various chemokine and Wnt receptors and ligands that may control cell polarization and directional migration. Moreover, we are also currently verifying and characterizing several additional gene families that are highly expressed in T cells after migration in response to or simply stimulation with CXCL12, CCL19, gp120 and HIV-1 virus. Moreover, the role of lipid rafts in chemokine biology and HIV infectivity are also under examination using microarray analysis. A greater understanding of the transcriptional signals differentially induced by the ligation of various chemokine receptors may provide a means to dissect the pathways by which these chemoattractants induce cell migration and activation as well as any host transcriptional signals important in HIV entry and replication.

趋化因子在体外和体内都能诱导和指导人和啮齿动物白细胞的黏附、趋化、激活和脱颗粒。CXCL12和CCL19是两种重要的趋化因子，在正常和炎症条件下调节T细胞的运动和激活。尽管有许多研究趋化因子功能的报道，但对其中涉及的转录事件知之甚少。在这里，我们对CXCL12处理的T细胞进行了微阵列分析，发现Wnt蛋白家族在CXCL12处理过程中显著上调。实时定量聚合酶链式反应和Western分析证实，在CXCL12刺激过程中，Wnt5A和其他非典型性Wnt途径成员的表达特异性上调，而-catenin和典型性Wnt家族成员的表达选择性下调。研究发现，WNT5A通过激活蛋白激酶C(PKC)，通过CXCL12-CXCR4轴增强信号传导。此外，我们的数据显示，在CXCL12的反应中，Wnt5A的表达是介导T细胞定向迁移所必需的，并且重组Wnt5A处理人T细胞会使T细胞对CXCL12诱导的迁移敏感。此外，CXCR4在转录和翻译上的持续表达也需要Wnt5A的表达。将EL4胸腺瘤转移作为T细胞迁移的模型，这些结果在体内得到进一步支持。综上所述，这些数据首次证明，Wnt5A是CXCL12-CXCR4信号和人类和小鼠T细胞迁移的关键介质。有趣的是，我们还发现Wnt10A在CCL19趋化和维持CCR7在T细胞上的表达中起作用。这些发现可能揭示了多种趋化因子与Wnt受体和配体之间的新的合作信号网络，该网络可能控制细胞极化和定向迁移。 此外，我们目前还在验证和鉴定另外几个基因家族，这些基因家族在T细胞对CXCL12、CCL19、gp120和HIV-1病毒的反应或简单刺激下迁移后高表达。此外，脂筏在趋化因子生物学和艾滋病毒感染性中的作用也正在通过微阵列分析进行检查。更好地理解不同趋化因子受体连接诱导的转录信号可能提供一种手段来剖析这些趋化因子诱导细胞迁移和激活的途径，以及任何在HIV进入和复制中重要的宿主转录信号。