Phenotypic and functional analysis of Dendritic cells in Liver

肝脏树突状细胞的表型和功能分析

基本信息

批准号：
07457083
负责人：
INABA Kayo
金额：
$ 4.74万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457083/
关键词：
dendritic cells liver cytokines maturation phenotype function MHCクラスII

项目摘要

Dendritic cells (DCs) are a system of potent antigen-presenting cells that are distributed in lymphoid and nonlymphoid tissues where they are usually distinguished by their morphology and class II antigen expression. DCs in peripheral nonlymphoid tissues are thought to be a relatively immature cell type that capture antigens and migrate to T areas of the draining lymphoid organs. However, heterogeneity of DCs in nonlymphoid tissues is not well known. Therefore, the aim of this project is further characterization of DCs in terms of phenotype, function and distribution in the liver.A procedure was developed to maximize the yield of viable, nonparenchymal hepatic leukocytes by collagenase digestion of perfused liver fragments followed by density centrifugation with Nyco Prep^<TM>. FACS analyzes on isolates showed that N418 (CD11c)^+ cells are categorized into three different subpopulations besed on the cell size and the expression of class II,CD86 and DEC-205. All of them were CD3^-, CD4^-, and CD8^-, but strongly expressed CD45 and moderate levels of CD11b and CD44. The most immature DCs were small in size and negative or very little for class II,but expressed low level of B220. The intensity of class II was shown to correlate to the cell size. The large Class II^<high> cells with dendritic shape primarily localized in the triads but not in sinus, and CD11c^+ cells in sinus were negative for class II and relatively small round cells in immunohistochemical observation. When even small CD11c^+ cells, however, were cultured in the presence of GM-CSF for a few days, they became larger and increased class II,CD86 and DEC-205 expression, and potent allostimulatory activity was detected. These observations indicate that developmentally different populations of DCs are present in the distinguished part of the liver.

树突状细胞（DCS）是一种有效的抗原细胞系统，分布在淋巴类和非淋巴组织中，通常以其形态和II类抗原表达来区分它们。外周非淋巴组织中的DC被认为是一种相对不成熟的细胞类型，可捕获抗原并迁移到排水淋巴机器人的T区域。然而，非淋巴组织中DC的异质性尚不清楚。因此，该项目的目的是根据表型，功能和分布在肝脏中的进一步表征。制定了A程序，以通过灌注肝脏碎片的胶原酶消化，然后以密度为偏心的胶原酶消化来最大程度地提高可行的，非帕克马尔肝肝白细胞的产量。 NYCO Prep^<tm>。对分离株的FACS分析表明，N418（CD11C）^+细胞被分为三种不同的亚群，对II，CD86和DEC-205的细胞大小和表达。它们都是CD3^ - ，CD4^ - 和CD8^ - ，但强烈表达的CD45和中等水平的CD11b和CD44。最不成熟的DC的大小较小，而II类较小，但表现出低水平的B220。证明了II类的强度与细胞大小相关。具有树突状形状的大型II^<高>细胞主要定位于三合会，但不在鼻窦中，而在免疫组织化学观察中，II类和相对较小的圆形细胞为阴性。然而，当在GM-CSF存在几天的情况下，即使小CD11C^+细胞也被培养时，它们变得更大并增加了II类，CD86和DEC-205表达，并检测到有效的同种异性活性。这些观察结果表明，在肝脏的特殊部分中存在开发不同的DC种群。