Characterization of CCL28,CXCL14, and CXCL17, three mucosal chemokines

三种粘膜趋化因子 CCL28、CXCL14 和 CXCL17 的表征

• 批准号: 8586295
• 负责人: Albert Zlotnik
• 金额: $ 37.61万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-05 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8586295
• 关键词: Address; Affect; Antibodies; Biological Assay; Bronchi; CXCL14 gene; Cells; Chemotaxis; Chlamydia; Data; Dendritic Cells; Development; Enzyme-Linked Immunosorbent Assay; Feces; Formalin; GPR2 gene; Generations; Genes; Homing; Human; IgA Deficiency; Immune response; Immune system; Immunity; Immunoglobulin A; Immunohistochemistry; Intestines; Knockout Mice; Lead; Lymphoid Tissue; Mammary gland; Mediating; Messenger RNA; Milk; Mucosal Immune Responses; Mucous Membrane; Mus; Oral cavity; Orphan; Paraffin Embedding; Pharmacologic Substance; Phenotype; Plasmablast; Play; Population; Predisposition; Production; Property; Proteins; Protocols documentation; Recruitment Activity; Role; Saliva; Salmonella; Sampling; Site; Staining method; Stains; Stem cells; T-Lymphocyte; TNFSF11 gene; Testing; Tissues; Trachea; cell motility; cell type; chemokine; chemokine receptor; cytokine; female reproductive system; gastrointestinal system; homologous recombination; human GPRC5C protein; human disease; insight; interleukin-21; microbicide; monocyte; mucosal site; novel; pathogen; receptor; release of sequestered calcium ion into cytoplasm; research study; response

DESCRIPTION (provided by applicant): The mucosal sites include the oral cavity, trachea and bronchi, the digestive system and the female reproductive system. The immune responses that develop in these sites are important to fend off potential pathogens. The chemokines are small secreted proteins that regulate the migration of cells of the immune system. We have identified three chemokines (CXCL14, CXCL17 and CCL28) as the most highly expressed in mucosal tissues. We hypothesize that they must have important functions in these mucosal sites. In Specific Aim 1, we will use study the mucosal immune system of mice genetically deficient for each of these chemokines in order to characterize their functions. We already have access to a CXCL14 (-/-) mouse, we can obtain an existing CCL28 (-/-) mouse (available from Deltagen) and we will produce a CXCL17 (-/-) from existing embryonal stem cells where the CXCL17 gene has been inactivated by homologous recombination (available through TIGM). We will also infect these mice with two bacterial pathogens (Chlamydia and Salmonella) to investigate whether these chemokines play a role in immunity against these pathogens. We have already observed that in the CXCL14 (-/-) mouse, the levels of IgA in the gut are severely reduced. Therefore, we hypothesize that CXCL14 is the main chemokine mediating the homing of IgA producing cells to the gut. In Specific Aim 2, we will study the mechanism through which CXCL14 controls IgA production in the gut. We will study whether the IgA-producing cells normally present in the gut exist in these mice. We will also study the M cells as well as the expression of other proteins that have been implicated in gut IgA responses (such as RANKL) as well as for the presence of other immune system cells known to exist in the mouse gut (Th17, Tregs). We will also study whether exogenously administered CXCL14 can reverse the lack of gut IgA in CXCL14 (-/-) mice. Two of these chemokines (CXCL14 and CXCL17) have no known receptors. Therefore, In Specific Aim 3, we will search for their receptors among various orphan G-Protein coupled receptors expressed in two cell types (immature dendritic cells and monocytes) that respond to these chemokines. We will screen by transfecting the GPCRs in appropriate host cells and detecting calcium fluxes specifically induced by the chemokines. Importantly, through these experiments we will identify a new chemokine receptor. Other chemokine receptors have been implicated in human diseases, and represent potential targets for pharmaceutical development. These results will lead to new insights into the role that these as yet unrecognized mucosal chemokines play in the development of mucosal immune responses.

描述（由申请方提供）：粘膜部位包括口腔、气管和支气管、消化系统和女性生殖系统。在这些部位产生的免疫反应对于抵御潜在的病原体非常重要。趋化因子是调节免疫系统细胞迁移的小分泌蛋白质。我们已经鉴定了三种趋化因子（CXCL 14、CXCL 17和CCL 28）在粘膜组织中表达最高。我们推测它们在这些粘膜部位一定具有重要的功能。在具体目标1中，我们将使用这些趋化因子中的每一种的遗传缺陷小鼠的粘膜免疫系统进行研究，以表征其功能。我们已经获得了CXCL 14（-/-）小鼠，我们可以获得现有的CCL 28（-/-）小鼠（可从Deltagen获得），我们将从现有的胚胎干细胞中产生CXCL 17（-/-），其中CXCL 17基因已通过同源重组失活（可通过TIGM获得）。我们还将用两种细菌病原体（衣原体和沙门氏菌）感染这些小鼠，以研究这些趋化因子是否在对这些病原体的免疫中发挥作用。我们已经观察到，在CXCL 14（-/-）小鼠中，肠道中伊加的水平严重降低。因此，我们假设CXCL 14是介导伊加产生细胞归巢到肠道的主要趋化因子。在具体目标2中，我们将研究CXCL 14控制肠道中伊加产生的机制。我们将研究正常存在于肠道中的IgA产生细胞是否存在于这些小鼠中。我们还将研究M细胞以及与肠道伊加反应有关的其他蛋白质（如RANKL）的表达，以及已知存在于小鼠肠道中的其他免疫系统细胞（Th 17，TcB）的存在。我们还将研究外源性给予CXCL 14是否可以逆转CXCL 14（-/-）小鼠中肠道伊加的缺乏。其中两种趋化因子（CXCL 14和CXCL 17）没有已知的受体。因此，在具体目标3中，我们将在两种细胞类型（未成熟树突状细胞和单核细胞）中表达的各种孤儿G蛋白偶联受体中寻找它们的受体，这些细胞对这些趋化因子有反应。我们将通过在适当的宿主细胞中检测GPCR并检测由趋化因子特异性诱导的钙流来进行筛选。重要的是，通过这些实验，我们将确定一个新的趋化因子受体。其他趋化因子受体与人类疾病有关，并代表了药物开发的潜在靶点。这些结果将导致新的见解，这些尚未认识到的粘膜趋化因子在粘膜免疫反应的发展中发挥的作用。