Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht University, Utrecht, Netherlands.
Chemical Biology and Drug Discovery Group, Department of Pharmacological Sciences, Utrecht University, Utrecht, Netherlands.
Front Immunol. 2023 Jan 9;13:1073034. doi: 10.3389/fimmu.2022.1073034. eCollection 2022.
The global demand of sustainable food sources leads to introduction of novel foods on the market, which may pose a risk of inducing allergic sensitization. Currently there are no validated assays mimicking the human mucosal immune system to study sensitizing allergenicity risk of novel food proteins. The aim of this study was to introduce a series of sequential human epithelial and immune cell cocultures mimicking key immune events after exposure to the common food allergen ovalbumin from intestinal epithelial cell (IEC) activation up to mast cell degranulation.
This human mucosal food sensitizing allergenicity model combines crosstalk between IEC and monocyte-derived dendritic cells (moDC), followed by coculture of the primed moDCs with allogenic naïve CD4+ T cells. During subsequent coculture of primed CD4+ T cells with naïve B cells, IgE isotype-switching was monitored and supernatants were added to primary human mast cells to investigate degranulation upon IgE crosslinking. Mediator secretion and surface marker expression of immune cells were determined.
Ovalbumin activates IEC and underlying moDCs, both resulting in downstream IgE isotype-switching. However, only direct exposure of moDCs to ovalbumin drives Th2 polarization and a humoral B cell response allowing for IgE mediated mast cell degranulation, IL13 and IL4 release in this sequential DC-T cell-B cell-mast cell model, indicating also an immunomodulatory role for IEC.
This coculture model combines multiple key events involved in allergic sensitization from epithelial cell to mast cell, which can be applied to study the allergic mechanism and sensitizing capacity of proteins.
全球对可持续食物来源的需求导致新型食物进入市场，这可能带来诱导过敏致敏的风险。目前，尚无模拟人体黏膜免疫系统的验证方法来研究新型食物蛋白的致敏性过敏原风险。本研究旨在介绍一系列连续的人上皮细胞和免疫细胞共培养物，模拟从肠上皮细胞（IEC）激活到肥大细胞脱颗粒的常见食物过敏原卵清蛋白暴露后的关键免疫事件。
该黏膜食物致敏性变应原模型结合了 IEC 和单核细胞衍生的树突状细胞（moDC）之间的串扰，随后将致敏的 moDC 与同种异体幼稚 CD4+T 细胞共培养。在致敏的 CD4+T 细胞与幼稚 B 细胞的随后共培养中，监测 IgE 同种型转换，并将上清液添加到原代人肥大细胞中，以研究 IgE 交联时的脱颗粒。测定免疫细胞的介质分泌和表面标志物表达。
卵清蛋白激活 IEC 和其下方的 moDC，均导致下游 IgE 同种型转换。然而，只有 moDC 直接暴露于卵清蛋白才能驱动 Th2 极化和体液 B 细胞反应，从而允许在这个顺序的 DC-T 细胞-B 细胞-肥大细胞模型中进行 IgE 介导的肥大细胞脱颗粒、IL13 和 IL4 释放，表明 IEC 还具有免疫调节作用。
该共培养模型结合了从上皮细胞到肥大细胞的过敏致敏的多个关键事件，可以应用于研究蛋白质的过敏机制和致敏能力。
