Lymphatic pumping and lymphatic transport of antigen to lymph node

淋巴泵和淋巴运输抗原至淋巴结

• 批准号: RGPIN-2015-03641
• 负责人: Liao, Shan
• 金额: $ 2.4万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=708051
• 关键词: Lymphatic; pumping; lymphatic; transport; antigen

Lymphatic vessels play essential roles in maintaining fluid balance and facilitating immune defense. While the heart beats to drive blood circulation, lymph transport primarily relies on lymphatic vessel pumping. Lymphatic pumping is driven by lymphatic smooth muscle cell (SMC) contraction. The function of lymphatic pumping is to propel lymph and cells transport to the draining lymph node (LN). When antigen and dendritic cells reach the draining lymph node (LN) through afferent lymphatic vessels, they must exit the vessels and migrate deep into the LN to activate T cell responses. However, it is not clear how changes in lymph flow may affect antigen delivery and antigen presentation in LN. In my previous studies, I have developed an intravital imaging model to study lymphatic pumping in mice. Using this model, I demonstrated that nitric oxide (NO) is required to maintain lymphatic pumping in physiological condition via the activation of the endothelial-derived nitric oxide synthase (eNOS). In contrast, the inducible nitric oxide synthase (iNOS) is responsible for the inhibition of lymphatic pumping in oxazolone skin sensitization induced inflammation (OX-inflammation). We hypothesize that NO regulates lymphatic pumping, which consequently modulates lymph flow velocity and flow pattern, and affect antigen delivery in lymph node. OBJECTIVES Objective 1. To determine how NO regulates lymphatic SMC to control lymphatic pumping. We will study if NO directly signals on SMCs to cause SMC relaxation or via indirectly interrupt SMC and lymphatic endothelial cell (LEC) interaction. We will use eNOS-/- mice, or broad NOS inhibitor L-NMMA or iNOS-/- mice to block NOSs and study how lymphatic SMCs are regulated in physiological and OX-inflammation.  Objective 2. To determine how lymphatic pumping controls the lymph flow velocity and flow pattern. We will measure the lymph flow in wild type, eNOS-/- and iNOS-/- mice in physiological and inflammatory conditions. We will study how lymph flow regulated LEC gene expression with cell culture using flow chamber to manipulate lymph flow velocity and flow pattern in variant conditions. Objective 3. To determine how lymphatic pumping manipulates antigen delivery to the draining LN. Free form of small antigens travel along the LN conduit system and are quickly captured by lymph node resident dendritic cells (DCs). We will use fluorescently labeled ovabulmin as small antigen to measure the antigen delivery in the draining LN when lymphatic pumping is impaired in variant conditions. We will study how these changes will alter the efficiency of antigen capture by LN DCs and initiation of T cell responses. The long-term goal of my studies will address fundamental questions of lymphatic pumping and lymphatic transport of antigen to the draining lymph node in order to efficiently present antigen and generate immune defense.

淋巴管在维持液体平衡和促进免疫防御方面起着至关重要的作用。虽然心脏跳动来驱动血液循环，但淋巴的运输主要依靠淋巴管的泵送。淋巴泵是由淋巴管平滑肌细胞(SMC)收缩驱动的。淋巴泵的功能是推动淋巴和细胞运输到引流淋巴结(LN)。当抗原和树突状细胞通过传入淋巴管到达引流淋巴结(LN)时，它们必须离开血管并深入LN以激活T细胞反应。然而，淋巴流量的改变如何影响LN的抗原递送和抗原递呈尚不清楚。在我之前的研究中，我开发了一个活体成像模型来研究小鼠的淋巴泵。使用这个模型，我证明了一氧化氮(NO)是通过激活内皮源性一氧化氮合酶(ENOS)来维持生理状态下的淋巴泵送的。相反，诱导型一氧化氮合酶(INOS)负责在恶唑酮皮肤致敏诱导的炎症(OX-炎症)中抑制淋巴泵送。我们推测，NO调节淋巴泵，进而调节淋巴流速和流型，从而影响淋巴的抗原递送。 目标 目的1.探讨一氧化氮(NO)对淋巴管SMC的调节作用。我们将研究SMC上是否没有直接信号导致SMC松弛，或者通过间接中断SMC与淋巴管内皮细胞(LEC)的相互作用。我们将使用eNOS-/-小鼠，或广泛的一氧化氮合酶抑制剂L-NMMA或iNOS-/-小鼠来阻断NOSS，并研究淋巴SMC如何在生理和牛炎症中进行调节。 目的2.确定淋巴泵是如何控制淋巴流速和流型的。我们将测量野生型、eNOS-/-和iNOS-/-小鼠在生理和炎症条件下的淋巴流量。我们将通过细胞培养来研究淋巴流动如何调节LEC基因的表达，利用流室在不同条件下操纵淋巴流动速度和流动模式。 目的3.确定淋巴抽吸如何操纵抗原递送至引流的LN。游离形式的小抗原沿LN管道系统传播，并迅速被淋巴结内树突状细胞(DC)捕获。我们将使用荧光标记的卵泡蛋白作为小抗原来测量在不同情况下淋巴泵功能受损时引流层神经中的抗原递送情况。我们将研究这些变化将如何改变LN DC捕获抗原的效率和启动T细胞反应。 我的研究的长期目标将解决淋巴泵送和淋巴运输抗原到引流淋巴结的基本问题，以便有效地呈递抗原和产生免疫防御。