Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells.
Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells.
复制标题
DOI：
10.1097/00007890-199408270-00015
复制
发表时间：
1994-08
期刊：
影响因子：
6.2
通讯作者：
J. Woo;Lina L. Lu;A. Rao;Youping Li;V. Subbotin;T. Starzl;A. Thomson
中科院分区：
文献类型：
--
作者：
J. Woo;Lina L. Lu;A. Rao;Youping Li;V. Subbotin;T. Starzl;A. Thomson
供体肝脏来源的树突状细胞（DC）最近在器官移植受体的各种淋巴和非淋巴组织中被发现，包括移植和永久接受主要组织相容性复合体（MHC）的非免疫抑制小鼠-不同的肝脏移植物。这些发现提出了关于肝脏耐受性基础的问题，特别是关于肝源性DC的特性。为了进一步研究正常小鼠（B10.BR;H-2k, I-Ek）肝脏中白细胞的结构、免疫表型和异源刺激活性，研究人员开发了一种方法，通过胶原酶消化灌注的肝碎片和密度离心（Percoll），最大限度地获得有活力的非实质细胞（NPC）。这些细胞包括表达淋巴细胞和髓细胞表面抗原的群体。与脾细胞相比，在原发性混合白细胞反应（MLR）中，它们被证明是初始（B10; H-2b, I-E-）脾T细胞的良好同种刺激物。在鼻咽癌过夜（18小时）孵育后，在甲硝唑胺梯度上富集短暂贴壁的低密度（LD）细胞，可以恢复少量细胞（约为1 / 3）。每个肝脏2-5 x 10(5)，其中许多表现出明显的DC形态。流式细胞术分析显示，这些细胞是CD3-、CD4-、CD8-和B220-，但强烈表达CD45（白细胞共同抗原），以及轻度至中度水平的CD11b、热稳定抗原和CD44。这些细胞也表达中等强度的NLDC 145，但不表达DC限制性标记33D1，这些标记已被证明在不同器官分离的小鼠DC上有差异表达。通过免疫细胞化学和流式细胞术检测，这个dc富集的群体比NPC具有更强的MHC II类(I-Ek)+，并且对幼稚T细胞表现出更强的同种异体刺激活性。这些发现表明，新鲜分离的小鼠肝脏NPC，可能与其原位的对应物，在过夜培养后，在非粘附的低密度（dc富集）部分中表现出增强的异源刺激活性。他们进一步表明，肝DC的成熟可能在决定肝同种异体移植物的免疫原性和/或耐受性方面起关键作用。
Donor liver-derived dendritic cells (DC) have recently been identified within various lymphoid and nonlymphoid tissues of organ allograft recipients, including nonimmunosuppressed mice transplanted with and permanently accepting major histocompatibility complex (MHC)-disparate hepatic allografts. These findings have raised questions about the basis of the tolerogenicity of the liver--and, in particular, about the properties of liver-derived DC. To study further the structure, immunophenotype and allostimulatory activity of leukocytes resident in normal mouse (B10.BR;H-2k, I-Ek) liver, a procedure was developed to maximize the yield of viable, nonparenchymal cells (NPC) obtained following collagenase digestion of perfused liver fragments and density centrifugation (Percoll). These cells comprised populations expressing lymphoid and myeloid cell surface antigens. As compared with spleen cells, they proved good allostimulators of naive (B10; H-2b, I-E-) splenic T cells when tested in primary mixed leukocyte reactions (MLR). After overnight (18-hr) incubation of the NPC, enrichment for transiently adherent, low-density (LD) cells on metrizamide gradients permitted the recovery of low numbers of cells (approx. 2-5 x 10(5) per liver), many of which displayed distinct DC morphology. Flow cytometric analysis revealed that these cells were CD3-, CD4-, CD8-, and B220-, but strongly expressed CD45 (leukocyte-common antigen), and mild-to-moderate levels of CD11b, heat-stable antigen, and CD44. The cells also expressed moderate intensity of NLDC 145 but not 33D1, DC restricted markers which have been shown to be differentially expressed on mouse DC isolated from various organs. This DC-enriched population was more strongly MHC class II(I-Ek)+ than NPC, as determined by immunocytochemistry and flow cytometry and exhibited much more potent allostimulatory activity for naive T cells. These findings demonstrate that freshly isolated murine liver NPC, and perhaps their counterparts in situ, exhibit allostimulatory activity that is enhanced in the non-adherent, low-density (DC-enriched) fraction after overnight culture. They further suggest that the maturation of liver DC may play a key role in determining the immunogenicity and or tolerogenicity of hepatic allografts.