Diversity of intraepithelial CD8aa T cells that recognize antignes from commensal flora

识别共生菌群抗原的上皮内 CD8aa T 细胞的多样性

• 批准号: 9413085
• 负责人: LESZEK IGNATOWICZ
• 金额: $ 18.94万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9413085
• 关键词: Address; Affect; Antigenic Specificity; Antigens; Area; Binding; Biomedical Research; Breeding; CD8B1 gene; Cell physiology; Cells; Clonal Expansion; Clone Cells; Code; Environment; Equilibrium; Future; Genetic Polymorphism; Germ-Free; Goals; Harvest; High-Throughput Nucleotide Sequencing; Human; Immune; In Situ; Individual; Intestines; Label; Lead; Lymphocyte; Methodology; Methods; Microbe; Mouse Strains; Mus; Organ; Peripheral; Phenotype; Physiological; Population; Proteins; Protocols documentation; Recovery; Recruitment Activity; Reporter; Research; Small Intestines; Specificity; Specimen; Staining method; Stains; Symbiosis; T-Cell Activation; T-Lymphocyte; Testing; Thymus Gland; Transcript; Wild Type Mouse; autoreactivity; commensal microbes; complementarity-determining region 3; cross reactivity; experimental study; gastrointestinal epithelium; germ free condition; in vivo; innovation; intestinal homeostasis; intraepithelial; microbial; microorganism antigen; mouse model; novel strategies; public health relevance; receptor; response; thymocyte

 DESCRIPTION (provided by applicant): The small intestine contains a significant number of intraepithelial T cells (IELs) that express CD8αα homodimer and αβTCRs. Currenlty these cells function and antigenic specificities remain enigmatic. Our long term goal is to understand how these cells contribute to homeostatic balance in the intestine, and how their TCR repertoires respond to microbial flora. Our central hypothesis is that clonal expansions and selective trophism of these IELs depends on microbial antigens that these cells recognize in vivo. These interactions may also contribute to sustain intestinal equilibrium. To test our hypothesis we propose two specific aims. First we will show how these IELs αβTCR repertoire changes in response to particular microbial species. We will examine this issue in a mouse model mice where T cells express semi diverse repertoire of TCRs and in vivo activated IELs are labeled with fluorescent protein (GFP). Limited diversity of TCRs allows to track individual IEL clones in response to different microbial species and organs using TCR CDR3 regions as clone-specific tags, whereas GFP allows to isolate IELs activated in their native environment. We expect to show that prime repertoire of IELs is broad, but become more oligoclonal following contacts with specific commensal antigens. In our Specific Aim 2 we propose to refine our new method to retrieve original pairs of αβTCRs sequences expressed by individual IELs for high-throughput sequencing (HTS), and use this approach to determine natural diversity of αβTCRs on IELs from wild type mice. This protocol is universal and in the future can be used to analyze native αβTCRs from humans.

 描述（由申请人提供）：小肠含有大量表达 CD8αα 同二聚体和 αβTCR 的上皮内 T 细胞 (IEL)。目前这些细胞的功能和抗原特异性仍然是个谜。我们的长期目标是了解这些细胞如何促进肠道内的稳态平衡，以及它们的 TCR 库如何对微生物菌群做出反应。我们的中心假设是，这些 IEL 的克隆扩增和选择性营养取决于这些细胞在体内识别的微生物抗原。这些相互作用也可能有助于维持肠道平衡。为了检验我们的假设，我们提出了两个具体目标。首先，我们将展示这些 IEL αβTCR 库如何响应特定微生物物种而变化。我们将在小鼠模型中研究这个问题，其中 T 细胞表达半多样化的 TCR 库，并且体内激活的 IEL 被荧光蛋白 (GFP) 标记。 TCR 的有限多样性允许使用 TCR CDR3 区域作为克隆特异性标签来跟踪单个 IEL 克隆以响应不同的微生物物种和器官，而 GFP 允许分离在其天然环境中激活的 IEL。我们希望证明 IEL 的主要功能很广泛，但在与特定共生抗原接触后变得更加寡克隆。在我们的具体目标 2 中，我们建议改进我们的新方法，以检索个体 IEL 表达的原始 αβTCR 序列对进行高通量测序 (HTS)，并使用这种方法来确定野生型小鼠 IEL 上 αβTCR 的自然多样性。该协议是通用的，将来可用于分析人类的天然 αβTCR。

项目成果

Non-canonicaly recruited TCRαβCD8αα IELs recognize microbial antigens.

• DOI: 10.1038/s41598-018-29073-7
• 发表时间: 2018-07-18
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Wojciech L;Szurek E;Kuczma M;Cebula A;Elhefnawy WR;Pietrzak M;Rempala G;Ignatowicz L
• 通讯作者: Ignatowicz L