Endocannabinoid regulation of host-helminth interaction

寄生蠕虫表现出非凡的能力，能够建立慢性的，通常是终生的， 通过触发多种机制来调节宿主的免疫反应。反过来， 宿主诱导平衡的免疫反应，在限制组织的同时介导蠕虫的杀灭 损坏。在这个多PI探索性方案中，我们确定内源性大麻素(ECB)系统为 这种动态宿主-蠕虫交互作用的一个以前未被认识到的贡献者。脂类 信号分子ECB是人体天然的类似大麻的分子，调节神经 上瘾和进食等行为。ECB的受体在整个细胞内表达 包括通过免疫细胞，其中ECB介导的信号可以抑制炎症。 以巴西拟圆线虫感染作为小鼠地蠕虫模型 感染，我们观察到在感染的人中有显著的ECB产生和ECB受体表达 在功能上影响宿主免疫反应和寄生虫卵负荷的组织。 此外，我们证明了NB在生命周期的每个阶段都会产生ECB，并识别了可能的基因 对于NB和寄生线虫基因组中的ECB合成和降解酶 感染人类。我们的中心假设是ECB在双向交流中起中介作用 在决定宿主生理、免疫反应和蠕虫的寄主和蠕虫之间 寄生。 在目标1中，我们将研究蠕虫感染诱导的内源性大麻素如何影响 通过(I)量化感染NB后的ECB水平和ECB信号；(Ii)废除ECB 药理试剂或eCBR缺陷小鼠的受体信号；(Iii)整合 用于识别与统计模型的功能交互的数据集。在目标2中，我们将使用分子 工具和高通量显微镜描绘寄生虫的内源性大麻素途径 蠕虫。我们将(I)利用药理学工具评估NB衍生的ECB和ECB信号 在NB中促进或取消ECB信号；以及(Ii)研究ECB在NB宿主中的作用 利用一种新的共培养系统进行免疫细胞相互作用。

Parasitic helminths exhibit the remarkable ability to establish chronic, often lifelong, infections by triggering multiple mechanisms to regulate the host immune response. In turn, the host induces a balanced immune response that mediates helminth killing while limiting tissue damage. In this multi-PI exploratory proposal, we identify the endocannabinoid (eCB) system as a previously unrecognized contributor to this dynamic host-helminth interaction. The lipid signaling molecules eCBs are the body’s natural cannabis-like molecules that regulate neural behaviors such as addiction and feeding. Receptors for eCBs are expressed throughout the body, including by immune cells, where eCB-mediated signaling can dampen inflammation. Following Nippostrongylus brasiliensis (Nb) infection as a mouse model of geohelminth infection, we observed significant eCB production and eCB receptor expression in the infected tissue that functionally impacted the host immune response and helminth egg burden. Moreover, we show that Nb produces eCBs at every life cycle stage, and identify putative genes for eCB synthetic and degradative enzymes in genome of Nb and parasitic nematodes that infect humans. Our central hypothesis is that eCBs mediate bi-directional communication between the host and helminth that dictates host physiology, immune response and helminth parasitism. In Aim 1 we will investigate how helminth infection-induced endocannabinoids affect the host by (i) quantifying eCB levels and eCB signaling following Nb infection; (ii) abrogating eCB receptor signaling by pharmacologic reagents or eCBR deficient mice; (iii) integrating the datasets to identify functional interactions with statistical models. In Aim 2 we will use molecular tools and high throughput microscopy to delineate the endocannabinoid pathway in the parasitic helminth. We will (i) evaluate Nb-derived eCBs and eCB signaling utilizing pharmacologic tools to promote or abrogate eCB signaling in Nb; and (ii) examine the role of eCBs in Nb-host immune cell interaction utilizing a novel co-culture system.