Control of placental infection by decidual NK cell secreted granulysin

蜕膜NK细胞分泌颗粒溶素控制胎盘感染

基本信息

批准号：
9236206
负责人：
Judy Lieberman
金额：
$ 21.63万
依托单位：
HARVARD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-04 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9236206
关键词：
Allogenic Alpha Cell Anti-Bacterial Agents Antimicrobial Effect Bacteria CD8B1 gene Cell Culture Techniques Cell Line Cell secretion Cells Cessation of life Conflict (Psychology)Cytoplasmic Granules Cytotoxic T-Lymphocytes Data Decidua Dependence Dose Enzymes Equilibrium Exocytosis Exposure to Fetal Distress Fetal Tissues Fetus Frequencies Gene Expression Goals Granzyme HLA G antigen Herpesviridae Human Human Parvovirus B19 Image Immune Immune Tolerance Immune response Immune system Immunity Infection Inflammatory Killer Cells Kinetics Lead Listeria monocytogenes Maternal Health Maternal-Fetal Exchange Mediating Membrane Microbe Modeling Molecular Natural Killer Cells Parasites Peptide Hydrolases Peptides Play Population Pregnancy Pregnancy Complications Pregnancy Trimesters Pregnant Uterus Premature Labor Process Protein Isoforms Proteins Resistance Role Rubella Side Site Specificity Streptococcus Group B Stromal Cells Synapses Syphilis Termination of pregnancy Tissues Toxoplasma gondii Vascular remodeling Virus antimicrobial peptide base cell killing congenital anomaly cytokine cytotoxic extracellular fetal fetus cell granulysin immunological synapse in utero insight killings microbial novel pathogen perforin peripheral blood placental infection public health relevance receptor function therapy development trophoblast

项目摘要

DESCRIPTION (provided by applicant): During pregnancy maternal immune cells need to establish immune tolerance of fetal and placental tissues, but also protect against infection. Decidual NK cells (dNK) are the largest maternal immune cell population at the maternal-fetal interface, where maternal and fetal tissues interact. This proposal will explore a novel and exciting mechanism by which dNK kill bacteria that infect fetal trophoblasts, but spare the fetal cells. Human dNKs highly express granulysin, an antimicrobial peptide that preferentially disrupts the membranes of microbes, but is less active against mammalian membranes. Granulysin is produced as an inactive 15 kDa pro-peptide that is processed in killer cell cytotoxic granules to a 9 kDa membranolytic peptide. Our preliminary data suggest that dNK cells have two intracellular pools of granulysin - one pool contains both the active and inactive forms of granulysin in cytotoxic granules together with perforin and granzymes, while the other pool contains only the inactive granulysin precursor. Preliminary data demonstrate that dNK constitutively secrete high levels of granulysin without the other cytolytic molecules. The culture supernatants from dNKs, but not peripheral blood NK cells (pNK), kill extracellular L. monocytogenes as well as L. monocytogenes within fetal trophoblast cell lines without killing the trophoblast. Based on these preliminary data, we hypothesize that granulysin secretion kills intracellular pathogens in fetal cells without harming those cells. To investigate this hypothesis, we propose to confirm our preliminary data showing that intracellular microbes are killed by dNK secretion of granulysin, independently of perforin and granzymes; determine whether bacteria or bacterial products or inflammatory cytokines upregulate granulysin expression and secretion; and understand how the trophoblast resists killing. We will also define whether granulysin secretion is constitutive or regulated and investigate whether dNKs respond differently to infected fetal extravillous trophoblasts (EVT, the most invasive fetal cells in the decidua) than t maternal decidual stromal cells (DSC). We will use imaging to determine whether dNK cells form an immune synapse with these infected cells, whether the type of synapse differs and whether the encounter triggers cytotoxic granule release. The models used will be 3 pathogens that cause complications during pregnancy - L. monocytogenes, Group B Streptococci (GBS) and Toxoplasma gondii.

描述（由适用提供）：在怀孕期间，孕产妇免疫小球需要建立胎儿和胎盘组织的免疫耐受性，但也可以防止感染。 decIDUAL NK细胞（DNK）是母体界面上母体和胎儿组织相互作用的母体免疫菌株种群。该建议将探索一种新颖而令人兴奋的机制，DNK杀死感染胎儿滋养细胞但要避免胎儿细胞的细菌。人DNK高度表达颗粒素，一种优先破坏微生物膜的抗菌肽，但针对哺乳动物膜的活性较低。颗粒素作为一种非活性15 kDa促肽产生，在杀手细胞的细胞毒性颗粒中加工至9 kDa膜 - 硝基化肽。我们的初步数据表明，DNK细胞具有两个细胞内颗粒蛋白 - 一个池在细胞毒性颗粒中含有活性和不活跃的颗粒素形式，以及颗粒蛋白和颗粒酶，而其他池仅包含无活性颗粒素前体。初步数据表明，DNK在没有其他细胞溶解分子的情况下组成性秘密高水平的颗粒素。文化来自DNK的上清液，但不是外周血NK细胞（PNK），杀死细胞外乳杆菌单核细胞增生李斯特菌以及胎儿滋养细胞细胞系中的单核细胞增生李斯特氏菌，而无需杀死滋养细胞。基于这些初步数据，我们假设颗粒状分泌杀死胎儿细胞中的细胞内病原体而不会损害这些细胞。为了研究这一假设，我们还将定义颗粒素的分泌是组成型还是受调节，并研究DNK对受感染的胎儿乳房滋养细胞的反应（EVT，决策中最具侵入性的胎儿细胞）与T孕产妇乳腺基质细胞（DSC）（DSC）。我们将使用成像来确定DNK细胞是否与这些感染细胞形成免疫，突触的类型是否有所不同，以及相遇是否触发了细胞毒性颗粒释放。所使用的模型将是3种妊娠期间引起并发症的病原体-L。单核细胞增生菌，B组链球菌（GBS）和弓形虫Gondii。