Pathways regulating plasmacytoid dendritic cells in Peyers Patches

派氏斑中浆细胞样树突状细胞的调节途径

• 批准号: 8432435
• 负责人: Stephanie S Watowich
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2014-08-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432435
• 关键词: Address; Animals; Antigen-Presenting Cells; Antigens; Antiviral Response; Biological Assay; Bone Marrow; CCR9 gene; CD4 Positive T Lymphocytes; CD8B1 gene; Cells; Characteristics; Dendritic Cells; Development; Disease; Environment; Equilibrium; Future; Generations; Goals; Homeostasis; Immune; Immune System Diseases; Immune Tolerance; Immune response; Immunity; In Vitro; Inflammation; Inflammatory; Interferon Type I; Interferons; Interleukin-17; Intestines; Investigation; Knowledge; Ligands; Lymphoid; Malignant Neoplasms; Measures; Modeling; Molecular; Mus; Natural Immunity; Organ; Pathway interactions; Phenotype; Play; Population; Production; Receptor Protein-Tyrosine Kinases; Regulatory T-Lymphocyte; Role; STAT1 gene; STAT3 gene; Signal Transduction; Spleen; Structure of aggregated lymphoid follicle of small intestine; Surface; T cell response; T-Lymphocyte; Testing; Toll-like receptors; Transplantation; Viral; adaptive immunity; conditioning; cytokine; gastrointestinal; immune activation; in vivo; novel; oral tolerance; pathogen; progenitor; reconstitution; research study; response; therapeutic development; therapy development; transcription factor

DESCRIPTION (provided by applicant): Plasmacytoid dendritic cells (pDCs) are principal type I interferon(IFN)-producing cells in bone marrow and lymphoid organs that are thought to play critical roles in anti-viral immunity. Upon activation through Toll-like receptors (TLRs), pDCs secrete IFN and mature to an antigen-presenting state in which they are able to elicit adaptive immune responses including CD4+ T cell polarization. The pDC developmental pathway proceeds through a common DC progenitor (CDP), which generates both pDCs and conventional DCs. PDC production depends on signaling by the receptor tyrosine kinase Flt3 and the transcription factors STAT3 and E2-2. A pDC subset was identified within Peyer's Patches (PP) in the gut that is unable to produce type I IFNs upon TLR triggering. This subset is implicated in oral tolerance; however the developmental origin and role for PP pDCs in eliciting adaptive immune responses are largely unknown. We found that PP pDC accrual required cell autonomous signaling via type I IFNs and STAT1, suggesting a distinct developmental or conditioning pathway compared to BM or splenic pDCs. Furthermore, we found that PP pDCs correlate with IL-17-secreting CD4+ T (Th17) cells in PPs. We hypothesize that PP pDCs arise from the same developmental pathway as pDCs localized in BM and spleen (i.e., via CDPs in response to Flt3-STAT3 signaling), however conditioning within the gut microenvironment may render distinct functional attributes including the ability to induce Th17 cell generation. We will test this hypothesis by investigating the pathways that regulate the accrual of pDCs within PPs (Aim 1). We will assess whether purified BM progenitors such as CDPs reconstitute PP pDCs in lethally irradiated mice or in a novel transplant model using Ifnar-/- animals that lack PP pDCs. Molecular pathways required for PP pDC generation will be examined by transplant experiments with BM progenitors from Stat1-/-, STAT3-deficient, Ifnar-/- or Tcf4-/- (E2-2-deficient) mice in steady state or in response to cytokines that elicit BM and spleen pDC generation (i.e., Flt3L, IFN-1). In Aim 2, we will determine whether PP pDCs are functionally distinct from BM or splenic pDCs in terms of eliciting CD4+ T cell responses, using in vitro CD4+ T cell polarization assays with purified PP, BM and splenic pDCs and naove CD4+ T cells. In addition, CD4+ T cell populations within PPs will be correlated with PP pDC reconstitution in Aim 1 to further examine the relationship between PP pDCs and Th cell responses, and the activation profiles of PP pDCs will be compared to BM and spleen pDCs by measuring candidate cytokine production, activating and inhibitory molecule expression and transcriptional profiles. Successful completion of this project will provide a mechanistic view of the PP pDC developmental pathway, and thus may contribute to development of therapeutic applications that enable redirection of the immune response in diseases associated with gastrointestinal inflammation.

描述（由申请人提供）：浆细胞样树突状细胞（pDC）是骨髓和淋巴器官中主要的 I 型干扰素（IFN）产生细胞，被认为在抗病毒免疫中发挥关键作用。通过 Toll 样受体 (TLR) 激活后，pDC 会分泌 IFN 并成熟至抗原呈递状态，在该状态下它们能够引发适应性免疫反应，包括 CD4+ T 细胞极化。 pDC 发育途径通过共同 DC 祖细胞 (CDP) 进行，该祖细胞产生 pDC 和常规 DC。 PDC 的产生取决于受体酪氨酸激酶 Flt3 以及转录因子 STAT3 和 E2-2 的信号传导。在肠道派尔氏集结 (PP) 内鉴定出 pDC 子集，该子集在 TLR 触发时无法产生 I 型 IFN。该子集与口服耐受有关；然而，PP pDCs 的发育起源和在引发适应性免疫反应中的作用在很大程度上尚不清楚。我们发现 PP pDC 的生成需要通过 I 型 IFN 和 STAT1 的细胞自主信号传导，这表明与 BM 或脾 pDC 相比，存在独特的发育或调节途径。此外，我们发现 PP pDC 与 PP 中分泌 IL-17 的 CD4+ T (Th17) 细胞相关。我们假设 PP pDC 与位于 BM 和脾脏的 pDC 源自相同的发育途径（即通过 CDP 响应 Flt3-STAT3 信号传导），然而肠道微环境内的调节可能会呈现不同的功能属性，包括诱导 Th17 细胞生成的能力。我们将通过研究调节 PP 内 pDC 增长的途径来检验这一假设（目标 1）。我们将评估纯化的 BM 祖细胞（例如 CDP）是否可以在致命辐射小鼠中或在使用缺乏 PP pDC 的 Ifnar-/- 动物的新型移植模型中重建 PP pDC。 PP pDC 生成所需的分子途径将通过来自 Stat1-/-、STAT3 缺陷、Ifnar-/- 或 Tcf4-/-（E2-2 缺陷）小鼠的 BM 祖细胞移植实验进行检查，这些小鼠处于稳态或对诱导 BM 和脾 pDC 生成的细胞因子（即 Flt3L、IFN-1）作出反应。在目标 2 中，我们将使用纯化的 PP、BM 和脾 pDC 以及 naove CD4+ T 细胞进行体外 CD4+ T 细胞极化测定，确定 PP pDC 在引发 CD4+ T 细胞反应方面是否在功能上不同于 BM 或脾 pDC。此外，PP 内的 CD4+ T 细胞群将与目标 1 中的 PP pDC 重建相关，以进一步检查 PP pDC 和 Th 细胞反应之间的关系，并且通过测量候选细胞因子的产生、激活和抑制分子表达以及转录谱，将 PP pDC 的激活谱与 BM 和脾 pDC 进行比较。该项目的成功完成将提供 PP pDC 发育途径的机制观点，因此可能有助于开发治疗应用，从而能够重定向与胃肠道炎症相关的疾病的免疫反应。

项目成果

Diversification of dendritic cell subsets: Emerging roles for STAT proteins.

• DOI: 10.4161/jkst.25112
• 发表时间: 2013-10-01
• 期刊: JAK-STAT
• 作者: Li HS;Watowich SS
• 通讯作者: Watowich SS

Assessing the development of murine plasmacytoid dendritic cells in Peyer's patches using adoptive transfer of hematopoietic progenitors.

• DOI: 10.3791/51189
• 发表时间: 2014-03-17
• 期刊: Journal of visualized experiments : JoVE
• 作者: Li HS;Watowich SS
• 通讯作者: Watowich SS