Gamma-Delta T cells and Cryptosporidiosis

Gamma-Delta T 细胞和隐孢子虫病

基本信息

批准号：
7414047
负责人：
Beth Diane Kirkpatrick
金额：
$ 13.46万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7414047
关键词：
Acute Adult Andro-Diane Animal Model Antigen-Presenting Cells Antigens Apoptosis Back Bangladesh Basic Science Biological Assay Bioterrorism CD4 Positive T Lymphocytes Cell Line Cells Centers for Disease Control and Prevention (U.S.)Child Chronic Class Clinical Country Cryptosporidiosis Cryptosporidium Cryptosporidium parvum Cytolysis Data Dependence Depth Diarrhea Disease Enteral Epithelial Cells Epithelium Glycoproteins Home environment Homing Human Immune response Immunity Immunology In Vitro Individual Infection Inflammation Inflammatory disease of the intestine Integrins Interleukin-2 International Intestinal Mucosa Intestines Knowledge Laboratories Lamina Propria Lead Ligands Link Lymphocyte MICA protein Mediating Mentors Modification Molecular Genetics Mucosal Immune Responses Newborn Animals Peripheral Population Prevention therapy Production Propionibacterium acnes Protozoa Recovery Research Research Personnel Research Proposals Role Sampling Source Stress Structure Surface T-Cell Receptor T-Cell Receptor gamma-Chain T-Lymphocyte Techniques Therapeutic Corynebacterium Parvum Thinking Training United States Up-Regulation Work age related cell killing cohort cytokine cytotoxic cytotoxicity experience in vivo intestinal epithelium intraepithelial investigator training killings neonate perforin peripheral blood professor programs receptor repaired response symposium

项目摘要

DESCRIPTION (provided by applicant): Cryptosporidium parvum is an important global cause of persistent and chronic diarrhea. In young children, particularly in underdeveloped countries, cryptosporidiosis is clinically more severe and is associated with more intestinal inflammation than infection in adults. Little is understood about the mucosal "innate" immune response to C. parvum. Gamma delta (gammadelta) T cells are an important T cell population in young children and in the intestinal tract. In response to infection at the intestine, T cells appear to expand in the periphery and selectively "home" back to the intestinal mucosa, via a gut-specific homing receptor (alpha4beta7). Intestinal gammadelta T cells appear to modulate recovery of the epithelium, respond to intestinal "stress" and may have important cytolytic function, especially in killing of infected intestinal epithelial cells (and possibly CD4+ alphabeta T cells). This K08 proposal seeks to train the investigator in classic and molecular/genetic immunology techniques and to apply this basic science knowledge to study mucosal immune responses to cryptosporidiosis in children, especially those in the developing world. The proposal hypotheses that gammadelta T cells are important in the innate immune response to cryptosporidiosis, are activated in response to C. parvum via the T cell receptor with necessary co-stimulation by the gut-specific MHC-I molecule (MIC-A), and kill infected epithelial cells by cytolysis and apoptosis. This information will then be applied to the study of gamma delta T cells from a small population of Bangladesh, children with acute cryptosporidiosis to demonstrate in vivo relevance. The primary mentor of this proposal is a full professor of Immunology with expertise in gammadelta T cells and with a laboratory skilled in the proposed research assays. Co-mentors are two experienced researchers, well known to the applicant, with depth in cryptosporidiosis and international research on enteric infections, respectively. The academic plan outlines structured formal coursework and conferences and provides depth to the laboratory training. In the research proposal, Aim 1 establishes whether the activation of the gammadelta cells is via the TCR and requires co-stimulation with the MHC-I like molecule MIC-A. In this aim, cloned gamma and gamma chains of the T cell receptor are transfected to a T cell receptor negative cell line and evaluated for function. Aim 2 evaluates the cytotoxicity of gammadelta via Fas-FasL and perforin and confirms whether cytotoxicity is antigen-dependent. Finally, to establish in vivo relevance, Aim 3 examines the gammadelta T cells from children with cryptosporidiosis, establishes whether these gammadelta cells (carrying the alpha4beta7 homing receptor) can be extracted from peripheral lymphocytes and if these clones have activation and cytolytic functions similar to the in vitro data. These data will advance our understanding of the mucosal immune response to cryptosporidiosis, which will lead to further advances in prevention and therapy.

描述（由申请人提供）：小隐孢子虫是全球持续性和慢性腹泻的重要原因。在幼儿中，特别是在不发达国家，隐孢子虫病在临床上更为严重，并且与成人感染相比，与更多的肠道炎症相关。人们对于微小念珠菌的粘膜“先天”免疫反应知之甚少。 γ δ (gammadelta) T 细胞是幼儿和肠道中重要的 T 细胞群。为了应对肠道感染，T 细胞似乎在外周扩张，并通过肠道特异性归巢受体 (alpha4beta7) 选择性地“归巢”回肠粘膜。肠道γδT细胞似乎可以调节上皮的恢复，对肠道“应激”做出反应，并且可能具有重要的细胞溶解功能，特别是在杀死受感染的肠道上皮细胞（可能还有CD4+ Alphata T细胞）方面。该 K08 提案旨在对研究人员进行经典和分子/遗传免疫学技术方面的培训，并应用这些基础科学知识来研究儿童（尤其是发展中国家的儿童）对隐孢子虫病的粘膜免疫反应。该提案假设，γδ T 细胞在对隐孢子虫病的先天免疫反应中很重要，通过 T 细胞受体在对小孢子虫的反应中被激活，并受到肠道特异性 MHC-I 分子 (MIC-A) 的必要共同刺激，并通过细胞溶解和凋亡杀死受感染的上皮细胞。然后，该信息将应用于来自孟加拉国一小部分患有急性隐孢子虫病的儿童的 γ δ T 细胞的研究，以证明体内相关性。该提案的主要导师是一位免疫学教授，拥有 γδ T 细胞方面的专业知识，并拥有擅长拟议研究测定的实验室。共同导师是两位经验丰富的研究人员，他们是申请人所熟知的，分别在隐孢子虫病和肠道感染的国际研究方面有深入的研究。学术计划概述了结构化的正式课程和会议，并为实验室培训提供了深度。在该研究提案中，目标 1 确定了 γδ 细胞的激活是否是通过 TCR 进行的，并且需要与 MHC-I 样分子 MIC-A 共同刺激。为此，将 T 细胞受体的克隆 γ 和 γ 链转染至 T 细胞受体阴性细胞系并评估其功能。目标 2 通过 Fas-FasL 和穿孔素评估 gammadelta 的细胞毒性，并确认细胞毒性是否具有抗原依赖性。最后，为了建立体内相关性，Aim 3 检查了隐孢子虫病儿童的 γδ T 细胞，确定这些γδ 细胞（携带 α4β7 归巢受体）是否可以从外周淋巴细胞中提取，以及这些克隆是否具有与隐孢子虫病相似的激活和溶细胞功能。体外数据。这些数据将增进我们对隐孢子虫病粘膜免疫反应的理解，从而推动预防和治疗的进一步进展。