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.

项目摘要 味蕾不断暴露于口服和食物中，但是三个途径 味觉细胞，口服微生物组和口服上皮免疫细胞之间的相互作用尚未是 进行了足够的详细研究。虽然口腔中的数百种微生物物种中的绝大多数是 对宿主无害或有益，口服微生物组的失调可能会引起味觉障碍。 在COVID-19，病毒肝炎，流感和念珠菌病的情况下观察到相关的味觉丧失， 列举一些例子。粘膜（例如肠道和扁桃体中的粘液）具有继发性淋巴组织 专门上皮植物上称为粘膜相关的淋巴组织（麦芽）称为卵泡相关 上皮（FAE）。 FAE包含透射蛋白酶的称为微膜细胞（M细胞）的免疫监视细胞 腔微生物并将其呈现给基础生发中心的免疫细胞，这会产生 适当的免疫反应。因此，M细胞是粘膜免疫的核心参与者，M的失调失调 已知细胞通路会引起感染。使用单细胞RNASEQ，我们发现了甜味受体 von ebner腺（VDC）的细胞（strc）和管道细胞，这是一种与味道相关的小唾液腺 乳头，表达几个M细胞标记基因，包括SPIB，M细胞所需的转录因子 发展与再生。这些发现已使用rnascope和双重标签确认 与STRC和其他味觉细胞类型标记基因的免疫组织化学。 RANKL的管理，增长 M细胞再生所需的因素导致MASTE乳头上M细胞标记基因的急剧上调 从野生型（WT），但不能击倒小鼠。我们假设STRC和VDC参与免疫 在味觉乳头上的监视方式与M细胞相同，并且该途径的扰动可能 导致感染，炎症和味觉丧失。我们将通过彻底研究 使用定量聚合酶链反应，RNASEQ和 组织学技术，并通过确定RANKL在特定中触发M细胞增殖的能力 无病原体（SPF）和无细菌（GF）WT小鼠和SPF尖峰有条件敲除小鼠（SPIBCKO小鼠）和 从中培养的味类动物。我们还将确定免疫细胞募集到味道的变化 上面描述的所有三种小鼠菌株/条件下的乳头。 Strc和VDC在体内和IN的能力 通过量化荧光的摄取，可以测量对透明腔腔微生物的味道器官 标记的纳米颗粒和表达大肠杆菌的绿色荧光蛋白。味道的变化 SPIBCKO小鼠的敏感性将使用阵风中的简短访问味觉测试确定。我们的研究 代表了味觉生物学和免疫之间的相交的一项新颖的尝试，并且有潜力 加深对味觉乳头状粘膜免疫监测的理解。成功完成 研究有望帮助设计新型策略，以治疗与感染，肥胖和衰老有关的味觉丧失。