Mucosal Immune Surveillance at the Taste Papillae

味乳头的粘膜免疫监视

• 批准号: 10693341
• 负责人: Sunil Kumar Sukumaran
• 金额: $ 15.55万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10693341
• 关键词: Advanced Development; Affect; Ageusia; Aging; Behavior; Behavioral Assay; Biological Assay; Biology; COVID-19; COVID-19 patient; Candida albicans; Candidiasis; Cell Proliferation; Cell physiology; Cells; Cilia; Development; Disease; Ductal Epithelial Cell; Epithelium; Escherichia coli; Exposure to; Food; Foundations; Future; Gene Expression; Gene Expression Profile; Genes; Germ-Free; Gland; Goals; Green Fluorescent Proteins; Growth Factor; Health; Histological Techniques; Homeostasis; Human; Immune; Immune response; Immunity; Immunoglobulin A; Immunohistochemistry; Immunologic Surveillance; Infection; Inflammation; Influenza; Intestines; Invaded; Kinetics; Knockout Mice; Label; Langerhans cell; Lymphoid Tissue; Maintenance; Measures; Mediating; Microbe; Microspheres; Minor salivary gland structure; Modeling; Mouse Strains; Mucosal Immunity; Mucous Membrane; Mucous body substance; Mus; Names; Natural regeneration; Obesity; Oral; Oral candidiasis; Oral cavity; Oral mucous membrane structure; Organoids; Pathology; Pathway interactions; Polymerase Chain Reaction; Population Heterogeneity; Receptor Cell; Role; Route; Signal Transduction; Structure of germinal center of lymph node; T-Lymphocyte; TNFSF11 gene; Taste Buds; Taste Disorders; Taste Perception; Testing; Tonsil; Transcriptional Regulation; Up-Regulation; Viral hepatitis; Wild Type Mouse; antimicrobial peptide; behavior measurement; cell regeneration; cell type; conditional knockout; cytokine; design; germ free condition; in vivo; intestinal epithelium; microbial; microbial colonization; microbiota; monocyte; mouse model; mucosa-associated lymphoid tissue; nanoparticle; novel; novel strategies; nutrition; opportunistic pathogen; oral cavity epithelium; oral microbial community; oral microbiome; recruit; secondary lymphoid organ; single-cell RNA sequencing; sweet taste perception; tongue dorsum; tongue papilla; transcription factor; transcriptome sequencing; transcytosis; uptake

Project Summary Taste buds are continually exposed to oral and food borne microbes, but the pathways underlying the three- way interactions between taste cells, the oral microbiome and oral epithelial immune cells have not been studied in sufficient detail. While the vast majority of the hundreds of microbial species in the oral cavity are either innocuous or beneficial to the host, dysregulation of the oral microbiome can cause taste disorders. Infection associated taste loss is observed in the cases of COVID-19, viral hepatitis, influenza, and candidiasis, to name a few examples. Mucosae such as those in the gut and tonsils possess secondary lymphoid tissues called mucosae associated lymphoid tissue (MALT) over laid by specialized epithelia call the follicle associated epithelium (FAE). FAE contain immune surveillance cells called microfold cells (M cells) that transcytose luminal microbes and present them to immune cells in the underlying germinal centers, that generate an appropriate immune response. Thus, M cells are central players in mucosal immunity, and dysregulation of M cell pathways are known to cause infection. Using single cell RNASeq, we discovered that sweet taste receptor cells (STRCs) and duct cells of the von Ebner gland (VDCs), a minor salivary gland associated with taste papillae, express several M cell marker genes, including Spib, a transcription factor required for M cell development and regeneration. These findings were confirmed using RNAScope and double label immunohistochemistry with STRC and other taste cell type marker genes. Administration of RANKL, a growth factor required for M cell regeneration led to dramatic upregulation of M cell marker genes in taste papillae from wild type (WT) but not Spib knock out mice. We hypothesize that STRCs and VDCs participate in immune surveillance at the taste papillae in the same manner as M cells, and that perturbances in this pathway might lead to infection, inflammation and taste loss. We will test this hypothesis by thoroughly examining the expression of M cell marker genes in taste papillae using quantitative polymerase chain reaction, RNASeq and histological techniques, and by determining the ability of the RANKL to trigger M cell proliferation in specific pathogen free (SPF) and germ free (GF) WT mice and SPF Spib conditional knockout mice (SpibCKO mice) and taste organoids cultured from them. We will also determine the changes in immune cell recruitment to the taste papillae in all three mouse strains/conditions described above. The ability of STRCs and VDCs in vivo and in taste organoids to transcytose luminal microbes will be measured by quantifying the uptake of fluorescently tagged nanoparticles and green fluorescent protein expressing Escherichia coli. The changes in taste sensitivity in SpibCKO mice will be determined using brief access taste test in a gustometer. Our study represents a novel foray into the intersection between taste biology and immunity and has the potential to deepen the understanding of mucosal immune surveillance at taste papillae. Successful completion of this study promises to help design novel strategies to treat taste loss associated with infection, obesity and aging.

项目概要 味蕾不断暴露于口腔和食源性微生物，但是这三种微生物的途径 味觉细胞、口腔微生物组和口腔上皮免疫细胞之间相互作用的方式尚未被研究 研究得足够详细。虽然口腔中数百种微生物物种中的绝大多数都是 无论对宿主无害还是有益，口腔微生物组的失调都会导致味觉障碍。 在 COVID-19、病毒性肝炎、流感和念珠菌病病例中观察到与感染相关的味觉丧失， 仅举几个例子。肠道和扁桃体等粘膜具有次级淋巴组织 称为粘膜相关淋巴组织（MALT），上面覆盖着特殊的上皮细胞，称为滤泡相关组织 上皮细胞（FAE）。 FAE 含有称为微折叠细胞（M 细胞）的免疫监视细胞，可转胞吞 腔内微生物并将它们呈现给底层生发中心的免疫细胞，从而产生 适当的免疫反应。因此，M 细胞在粘膜免疫和 M 细胞失调中发挥着核心作用。 已知细胞途径会引起感染。使用单细胞 RNASeq，我们发现甜味受体 埃布纳腺细胞 (STRC) 和管细胞 (VDC)，与味觉相关的小唾液腺 乳突，表达多种 M 细胞标记基因，包括 Spib，M 细胞所需的转录因子 发展和再生。这些发现已通过 RNAScope 和双标记得到证实 使用 STRC 和其他味觉细胞类型标记基因进行免疫组织化学分析。 RANKL 的管理，一个增长 M 细胞再生所需的因子导致味觉乳头中 M 细胞标记基因的急剧上调 来自野生型 (WT) 但不是 Spib 敲除小鼠。我们假设 STRC 和 VDC 参与免疫 以与 M 细胞相同的方式对味觉乳头进行监视，并且该通路中的扰动可能 导致感染、炎症和味觉丧失。我们将通过彻底检查来检验这个假设 使用定量聚合酶链反应、RNASeq 和 M 细胞标记基因在味乳头中的表达 组织学技术，并通过确定 RANKL 在特定情况下触发 M 细胞增殖的能力 无病原体（SPF）和无菌（GF）WT小鼠和SPF Spib条件敲除小鼠（SpibCKO小鼠）和 品尝从它们培养的类器官。我们还将根据口味确定免疫细胞募集的变化 上述所有三种小鼠品系/条件中的乳突。 STRC 和 VDC 在体内和体内的能力 将通过量化荧光的摄取来测量味觉类器官转胞吞腔内微生物的能力 标记的纳米颗粒和表达大肠杆菌的绿色荧光蛋白。口味的变化 SpibCKO 小鼠的敏感性将通过味觉计中的简短访问味觉测试来确定。我们的研究 代表了味觉生物学和免疫之间交叉点的新尝试，并且有潜力 加深对味觉乳头粘膜免疫监视的理解。顺利完成本次 研究有望帮助设计新的策略来治疗与感染、肥胖和衰老相关的味觉丧失。