Mechanisms of cetylpyridinium chloride inhibition of immune cell function

氯化十六烷基吡啶抑制免疫细胞功能的机制

基本信息

批准号：
10513855
负责人：
Julie Ann Gosse
金额：
$ 42.64万
依托单位：
UNIVERSITY OF MAINE ORONO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-11 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10513855
关键词：
Acute Affect Agriculture Anti-Bacterial Agents Antigens Binding Proteins Biochemical Biochemistry Biological Assay Cell Death Cell Degranulation Cell Surface Receptors Cell membrane Cell model Cell physiology Cell surface Cells Cetylpyridinium Chloride Charge Chemicals Clinical Data Color Confocal Microscopy Cytoplasmic Granules Cytosol Data Defect Dose Electrostatics Elements Endoplasmic Reticulum Enzymes Eukaryota Event Exocytosis Exposure to Fluorescence Fluorescence Microscopy Food Generations Golgi Apparatus Head Health Histamine Human IgE Image Immune Immune signaling Immune system Impairment In Vitro Inositol Investigation Lead Leukotrienes Lipids Literature MARCKS gene Measurement Measures Membrane Potentials Micelles Microscopy Microtubule Polymerization Microtubules Mitochondria Molecular Movement Nervous system structure Oral Oral mucous membrane structure PLC gamma1 Pathway interactions Persons Pharmacology Phosphatidylinositol 4,5-Diphosphate Phosphatidylinositols Phospholipase D Phosphorylation Physiological Plant Roots Production Property Prostaglandins Protein Isoforms Protein Kinase C Proteins Public Health Publications Reactive Oxygen Species Reader Research Resolution SNAP receptor Saliva Self Care Serotonin Signal Pathway Signal Transduction Signaling Protein T-Cell Receptor T-Lymphocyte Tail Testing Toxicology Tubulin Tyrosine Phosphorylation Workplace antimicrobial biochemical tools biophysical tools cell type crosslink cytokine density experimental study functional outcomes graduate student immune function insight mast cell nanoscale personal care products photoactivation receptor saturated fat synaptotagmin syntaxin transcriptome undergraduate student

项目摘要

People are widely exposed to high (mM) doses of the positively-charged antibacterial agent cetylpyridinium chloride (CPC) via janitorial and personal care products and foods treated with CPC, yet little is known about its toxicology in humans, especially below the critical micelle concentration (~900 µM). While applied CPC is retained in the oral mucosa and is released into saliva such that low-µM CPC continuously bathes oral cells, there is a dearth of publications on the effects of CPC on eukaryotes. The Gosse lab has discovered that exposure to non- cytotoxic, low-µM CPC concentrations potently inhibits signaling of mast cells, key players in the immune and nervous systems that share core signaling elements with T cells and other cell types. In mast cells, upon antigen crosslinking of cell surface receptors, a tyrosine phosphorylation cascade ensues, leading to activation of PLCγ1 enzymatic cleavage of phosphatidylinositol 4,5-bisphosphate (PIP2) and subsequent Ca2+ mobilization and degranulation: microtubule transport of granules to the cell surface, leading to exocytosis of bioactive substances such as histamine and serotonin. Analogous aggregation of T cell receptors leads also to tyrosine phosphorylation, Ca2+ mobilization, and downstream T cell function. CPC effects on both mast and T cell function will be determined. Preliminary data have led to the hypothesis that CPC inhibits immune cell function by electrostatically interfering with phosphorylation and PIP2, leading to displacement of PIP2-binding proteins, disrupted nanoscale clustering of PIP2, muted release of Ca2+ from endoplasmic reticulum (ER) stores, and, thereby, inhibited inflow of Ca2+ to the cytosol and extinguished microtubule polymerization. CPC effects on phosphorylation will be assessed by multiple means. Confocal microscopy and plate reader experiments will define CPC effects on sub-cellular localization and function of PLCγ1; of PLCγ1 product inositol 1,4,5-trisphosphate which initiates release of ER Ca2+; of Ca2+ dynamics in ER, mitochondria, Golgi, and cytosol; and of key elements downstream of Ca2+ including protein kinase C, phospholipase D, and microtubules. Whether CPC directly displaces PIP2 from its partner proteins will be measured. Super-resolution fluorescence photoactivation localization microscopy will interrogate nanoscale CPC effects on PIP2 clusters and other protein interactions crucial to immune function, including co-localization of Git1 regulator with tubulin as well as PIP2 with machinery required for granule exocytosis. This research will uncover the mechanisms underlying CPC disruption of immune cell function in order to fulfill an urgent need by providing insights into CPC effects on environmental human health.

人们广泛接触高（mM）剂量的带正电荷的抗菌剂氯化十六烷基吡啶 (CPC)，通过清洁和个人护理产品以及经过处理的食品 CPC，但对其对人体的毒理学知之甚少，尤其是在临界胶束以下浓度（~900 µM），而应用的 CPC 保留在口腔粘膜中并释放到口腔粘膜中。唾液中低 µM CPC 持续沐浴口腔细胞，但目前缺乏这方面的出版物 Gosse 实验室发现 CPC 对真核生物的影响。细胞毒性、低 µM CPC 浓度可有效抑制肥大细胞的信号传导，而肥大细胞是免疫和神经系统与 T 细胞和其他细胞共享核心信号元件在肥大细胞中，在细胞表面受体的抗原交联后，产生酪氨酸。随后发生磷酸化级联反应，导致 PLCγ1 酶促裂解的激活磷脂酰肌醇 4,5-二磷酸 (PIP2) 和随后的 Ca2+ 动员和脱颗粒：微管将颗粒转运至细胞表面，导致胞吐作用生物活性物质，如组胺和血清素，T 细胞的类似聚集。受体还导致酪氨酸磷酸化、Ca2+ 动员和下游 T 细胞初步数据将确定 CPC 对肥大细胞和 T 细胞功能的影响。导致了这样的假设：CPC 通过静电干扰抑制免疫细胞功能磷酸化和 PIP2，导致 PIP2 结合蛋白的置换，破坏 PIP2 的纳米级聚集，内质网 (ER) 库中 Ca2+ 的缓慢释放，从而抑制 Ca2+ 流入细胞质并消除微管聚合。 CPC 对磷酸化的影响将通过多种方法进行评估。读板器实验将确定 CPC 对亚细胞定位和功能的影响 PLCγ1；PLCγ1产物肌醇1,4,5-三磷酸，启动ER Ca2+的释放；内质网、线粒体、高尔基体和细胞质以及 Ca2+ 下游关键元素的动态；包括蛋白激酶C、磷脂酶D、微管是否直接取代CPC。来自其伙伴蛋白的 PIP2 将被测量。定位显微镜将探究纳米级 CPC 对 PIP2 簇和其他物质的影响对免疫功能至关重要的蛋白质相互作用，包括 Git1 调节器与微管蛋白以及 PIP2 与颗粒胞吐作用所需的机制。揭示 CPC 破坏免疫细胞功能的机制，以实现迫切需要提供有关 CPC 对环境人类健康影响的见解。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pharmaceutical agent cetylpyridinium chloride inhibits immune mast cell function by interfering with calcium mobilization.

药剂氯化十六烷基吡啶通过干扰钙动员来抑制免疫肥大细胞功能。

DOI：
10.1016/j.fct.2023.113980
发表时间：
2023
期刊：
Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association
影响因子：
0
作者：
Obeng,Bright;Potts,ChristianM;West,BaileyE;Burnell,JohnE;Fleming,PatrickJ;Shim,JuyoungK;Kinney,MarissaS;Ledue,EmilyL;Sangroula,Suraj;BaezVazquez,AlanY;Gosse,JulieA
通讯作者：
Gosse,JulieA