Modulation of conjunctival goblet cell differentiation by immunoregulatory cells

免疫调节细胞对结膜杯状细胞分化的调节

基本信息

批准号：
7445852
负责人：
CINTIA S. DE PAIVA
金额：
$ 19.19万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7445852
关键词：
Adenovirus Vector Antibodies Biological Assay Biopsy Boxing C57BL/6 Mouse CD4 Positive T Lymphocytes CD8B1 gene Cell Density Cell Differentiation process Cells Conjunctival Epithelium Development Disease Dominant-Negative Mutation Epithelial Eye Fibroblast Growth Factor Receptor 2 Goals Goblet Cells Grant Growth Factor Homeostasis Immune Immune system Immunoglobulin Variable Region Immunohistochemistry In Vitro Inflammation Inflammatory Interferon Type II Interferons Interleukins Knock-out Label Lead Letters Lymphocyte Lymphocyte Subset MUC5AC gene Metaplasia Mucins Mus Pathway interactions Phenotype Production Proline Proteins Research Proposals Role Skin Stress Surface T-Cell Receptor T-Lymphocyte Time Tissues Transforming Growth Factor beta Transforming Growth Factors Transgenic Mice conjunctiva cytokine density eye dryness insight intraepithelial keratinocyte growth factor keratinocyte growth factor receptor migration novel ocular surface overexpression prevent public health relevance receptor research study response

项目摘要

DESCRIPTION (provided by applicant): Modulation of conjunctival goblet cell differentiation by immunoregulatory cells: This project will investigate the role of local immunoregulatory pathways on conjunctival epithelial homeostasis and the response of the conjunctival epithelium to the desiccating stress of dry eye. We hypothesize that dry eye causes a decrease in the number of CD8+ and 34T intraepithelial lymphocytes (IEL) in the conjunctiva, resulting in decreased levels of immunoregulatory cytokines (e.g. TGF-22) and goblet-cell supporting factors (e.g. keratinocyte growth factor). This facilitates migration of interferon-gamma (IFN-?) producing CD4+ T cells into the conjunctival epithelium resulting in a cytokine imbalance that favors cornified envelope production, rather than goblet cell differentiation by the conjunctival epithelium. In Aim 1, the predominant subset of 34T cells in conjunctival biopsies from normal mouse eyes will be characterized by immunostaining and following in vitro expansion, the cytokine and growth factor profile of 34 and CD8 IEL will be evaluated by a multiplex immunobead assay, real-time PCR and microarray. The objective of Aim 2 is to investigate the role of IEL on survival of conjunctiva goblet cells after desiccating stress. To accomplish this, the resident density of CD8+ and 34 IELs will be evaluated prior to and after 5 and 10 days of experimental dry eye in C57BL/6 mice and in TCR-/- mice (34 T cell knock-out), CD8-/- mice and dominant negative fibroblast growth factor receptor 2 (dnFGFR2) transgenic mice. The time course of expression of KGF and TGF-22 will be evaluated. Co-localization of these factors to 34T and CD8+ T cells will be accomplished by dual label immunohistochemistry. The consequences of loss of 34T and CD8+ T cells and KGF and TGF-22 on conjunctival homeostasis will be confirmed by evaluating goblet cell and CD4+ cell density and expression of the goblet cell mucin MUC5AC and metaplasia marker small proline rich residue 2 protein (SPRR-2) in conjunctival sections from normal and dry eye mice. KGF and TGF-22 will be overexpressed using adenoviral vectors with the intent of preventing the goblet cell loss that occurs in dry eye. PUBLIC HEALTH RELEVANCE. This proposal will investigate the function of immuno-regulatory cells in the conjunctiva of the eye surface with regard to maintaining homeostasis and responding to desiccating environmental stress. These studies will provide new information about natural mechanisms to control inflammation on the ocular surface and may provide new insight into treating ocular surface inflammatory diseases.

描述（由申请人提供）：通过免疫调节细胞对结膜的杯状细胞分化的调节：该项目将研究局部免疫调节途径对结膜上皮稳态的作用，以及结膜上皮对干眼应激的结合上皮的反应。我们假设干眼症会导致结膜中的CD8+和34T上皮内淋巴细胞（IEL）的数量减少，从而导致免疫调节细胞因子（例如TGF-22）和Goblet-Cell-Cell-Cell-Cell-Cell-Cell-Cell-Cell-Cell-Cell-cell辅助因子（例如Keratinocyte的生长因子）。这有助于产生CD4+ T细胞的干扰素γ（IFN-？）迁移到结膜上皮，从而导致细胞因子失衡，从而有利于玉米包膜的产生，而不是通过造型上皮的杯状细胞分化。在AIM 1中，来自正常小鼠眼睛的结膜活检中的34T细胞的主要子集的特征是免疫染色和随后的体外膨胀，将通过多重Immunobead分析，实时PCR和MicroRay评估34和CD8 IEL的细胞因子和34和CD8 IEL的生长因子谱。 AIM 2的目的是研究IEL在干燥压力后的结膜杯细胞存活中的作用。为此，将在C57BL/6小鼠和TCR - / - 小鼠（34 T细胞敲除），CD8 - / - 小鼠和主导的负成纤维细胞生长因子受体2（DNFGFR2）转晶剂中，在C57BL/6小鼠和TCR - / - 小鼠（34 T细胞敲除）中，将评估CD8+和34 IEL的驻留密度。 KGF和TGF-22的表达时间过程将进行评估。这些因素将这些因素共定位到34T和CD8+ T细胞将通过双重标记免疫组织化学来实现。将通过评估杯状细胞和CD4+细胞密度以及Goblet细胞粘液MUC5AC的go和CD4+细胞密度以及分类标志物小脯氨酸2蛋白质2蛋白质2蛋白质（Sprrr-2）在Conjuntionction conjuntional sycect and normasions和Dry Dryecect中，通过评估Goblet细胞和CD4+细胞密度和表达来确认，34T和CD8+ T细胞以及KGF和TGF-22损失对结膜稳态的后果。 KGF和TGF-22将使用腺病毒载体过表达，以防止在干眼症中发生的杯状细胞损失。公共卫生相关性。该建议将研究眼表面结膜中免疫调节细胞在维持稳态和响应干燥环境应激方面的功能。这些研究将提供有关控制眼表炎症的自然机制的新信息，并可能为治疗眼表面炎症性疾病提供新的见解。