Bidirectional crosstalk between T lymphocytes and airway smooth muscle

T淋巴细胞和气道平滑肌之间的双向串扰

• 批准号: RGPIN-2017-05014
• 负责人: Martin, James
• 金额: $ 2.48万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=685176
• 关键词: Bidirectional; crosstalk; between; lymphocytes; airway

Contact of T cells with airway smooth muscle (ASM) cells triggers its growth and in turn ASM enhances T cell survival and proliferation. Interfering with contact augments T cell apoptosis and also interferes with ASM proliferation. A novel method of communication between the cells is through nanotubes, thin cellular projections from T cells to ASM. We demonstrated that anti-apoptotic factors such as Bcl2 and Mcl1 were present within the nanotubes and we have observed mitochondrial transfer from ASM to T cells. We have demonstrated a role for basic fibroblast growth factor-2 (bFGF-2) in nanotube formation (Appendix 4). However we know little of the biological consequences of communication via nanotubes between these cell types. The general hypothesis is that CD4+ T cells communicate with ASM cells via nanotubes that affect both ASM and T cell biology. To understand the biological consequences of nanotube formation between T cells and ASM cells, the drivers of the nanotube formation and the alterations in properties of the cells consequent upon this intercellular communication we will pursue the following general areas of research:****Aim 1: To determine the structure of the contact area of T cell nanotubes with ASM cells****Whether the T cell nanotube connection with ASM is similar to an immunological synapse will be investigated in this aim by examining the junction for the presence of the T cell receptor, MHC class II molecules, CD44, and co-stimulatory molecules.****Aim 2: To determine the role of growth factors in human CD4 T cell-driven ASM proliferation and in T cell activation and proliferation****The role basic fibroblast growth factor-2 and epidermal growth factor receptor ligands have in ASM proliferation and T cell activation and proliferation when T cells and ASM are in contact will be examined.****Aim 3: To explore the significance of mitochondrial transfer from ASM to T cells for T cell survival and function****We will test the effects of the inhibition of the movement of mitochondria from ASM to T cells via nanotubes by inhibiting motor proteins on T cell survival, metabolism and proliferation. We will nanofabricate culture plates to allow ASM and T cells to be cultured separately but with intervening channels to permit nanotubes to be formed between the cells.****Aim 4: To determine the effects of T cell contact with ASM cells on ASM contractile properties****Histamine induced calcium signals within ASM cells are reduced when in contact with T cells. We will examine the underlying molecular mechanisms. In addition we will explore the effects of the inhibition of nanotube formation on calcium signals by administering FGFR1 and EGFR inhibitors and anti-CD44 in concentrations that do not induce T cell apoptosis.****Anticipated results: We anticipate that we will elucidate the biological significance of cell to cell communication mediated by contact-dependent mechanisms between T cells and ASM cells.******

T细胞与气道平滑肌（ASM）细胞的接触触发其生长，并且反过来ASM增强T细胞存活和增殖。 干扰接触增强T细胞凋亡，也干扰ASM增殖。细胞之间的一种新的通信方法是通过纳米管，从T细胞到ASM的薄细胞突起。我们证明了抗凋亡因子如Bcl 2和Mcl 1存在于纳米管内，并且我们观察到线粒体从ASM转移到T细胞。我们已经证明了碱性成纤维细胞生长因子-2（bFGF-2）在纳米管形成中的作用（附录4）。 然而，我们对这些细胞类型之间通过纳米管进行通信的生物学后果知之甚少。一般的假设是，CD 4 + T细胞通过影响ASM和T细胞生物学的纳米管与ASM细胞通信。为了了解T细胞和ASM细胞之间纳米管形成的生物学后果，纳米管形成的驱动因素以及这种细胞间通讯导致的细胞性质的改变，我们将进行以下一般研究领域：* 目标1：为了确定T细胞纳米管与ASM细胞的接触区域的结构 **** T细胞纳米管与ASM的连接是否类似于免疫突触，将通过检查T细胞受体，MHC II类分子，CD 44和共刺激分子的存在来研究。目标二：为了确定生长因子在人CD 4 T细胞驱动的ASM增殖中以及在T细胞活化和增殖中的作用 * 将检查当T细胞和ASM接触时碱性成纤维细胞生长因子-2和表皮生长因子受体配体在ASM增殖以及T细胞活化和增殖中的作用。*目标三：为了探索线粒体从ASM转移到T细胞对于T细胞存活和功能的意义 * 我们将测试通过抑制马达蛋白来抑制线粒体经由纳米管从ASM到T细胞的运动对T细胞存活、代谢和增殖的影响。我们将纳米制造培养板，以允许ASM和T细胞分开培养，但中间有通道，以允许在细胞之间形成纳米管。目标4：为了确定T细胞与ASM细胞接触对ASM收缩特性的影响，当与T细胞接触时，ASM细胞内组胺诱导的钙信号减少。我们将研究潜在的分子机制。此外，我们将通过以不诱导T细胞凋亡的浓度施用FGFR 1和EGFR抑制剂以及抗CD 44来探索纳米管形成对钙信号的抑制作用。预期结果：我们预计，我们将阐明T细胞和ASM细胞之间的接触依赖性机制介导的细胞间通讯的生物学意义。