Functional characterization of adherent synovial fluid cells in rheumatoid arthritis - Destructive potential in vitro and in vivo
Functional characterization of adherent synovial fluid cells in rheumatoid arthritis - Destructive potential in vitro and in vivo
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DOI：
10.1002/art.11166
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发表时间：
2003-07-01
影响因子：
--
通讯作者：
Gay, S
中科院分区：
文献类型：
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作者：
Neidhart, M;Seemayer, CA;Gay, S
Objective.研究类风湿性关节炎（RA）患者滑膜液中粘附细胞的形态学和免疫学特征，探讨其在体外和体内的潜在功能。从RA患者和对照组中获得的滑膜液贴壁细胞的特征在于免疫组织化学，流式细胞术和电子显微镜。在体外，这些细胞在软骨颗粒的存在下培养。通过硫酸化糖胺聚糖（sGAG）释放到培养基中来监测软骨的破坏，并通过酶联免疫吸附测定法测量细胞培养上清液中基质金属蛋白酶1（MMP-1）的水平。为了抑制体外软骨破坏，在该系统中测试了MMP抑制剂马立马司他。在体内，在SCID小鼠共植入模型中，RA滑液贴壁细胞和RA滑膜成纤维细胞（作为阳性对照）与人软骨共植入肾包膜下，并在那里维持60天。滑膜液贴壁细胞由2个亚群组成，即大圆形巨噬细胞样细胞（CD 68+）和梭形成纤维细胞样细胞（Thy-1+）。当传代时，后者细胞增殖并将自身组织成三维结构。这使得它们能够到达用琼脂糖固定的胶原颗粒。来自滑膜组织的成纤维细胞不能用于本试验，因为它们仅在单层中生长，而不在琼脂糖上生长。大部分（>90%）传代的RA滑液贴壁细胞表达Thy-1+、CD 45-、CD 68-、CD 86-表型。电子显微镜检查未发现两种类型的成纤维细胞（滑膜组织或滑液）之间存在重要的形态学差异。然而，滑液粘附细胞表达较低水平的粘附分子，包括CD 54和半乳糖凝集素3，以及补体调节分子CD 55。与阴性对照细胞相比，RA滑液贴壁细胞中与细胞活性相关的sGAG体外释放高2.5倍。sGAG的释放与MMP-1的浓度相关，并以剂量依赖性方式被广谱MMP抑制剂marimastat抑制。RA滑膜液贴壁细胞与软骨共植入SCID小鼠后，表现出与组织来源的RA滑膜成纤维细胞相同的侵袭行为。类似于组织来源的RA滑膜成纤维细胞，RA滑液贴壁细胞，其中包含“浮动”锚定独立的成纤维细胞样细胞，介导软骨破坏的增生性滑膜组织独立。
Objective. To characterize the morphologic and immunologic features of adherent synovial fluid cells derived from patients with rheumatoid arthritis (RA), and to explore their potential function in vitro and in vivo by focusing on cartilage destruction.Methods. Synovial fluid adherent cells obtained from patients with RA and from control subjects were characterized by immunohistochemistry, flow cytometry, and electron microscopy. In vitro, these cells were cultured in the presence of cartilage particles. Cartilage destruction was monitored by the release of sulfated glycosaminoglycans (sGAG) into the medium, and the level of matrix metalloproteinase 1 (MMP-1) in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. To inhibit cartilage destruction in vitro, the MMP inhibitor marimastat was tested in this system. In vivo, in the SCID mouse coimplantation model, RA synovial fluid adherent cells and RA synovial fibroblasts (as positive controls) were coimplanted with human cartilage under the kidney capsule and maintained there for 60 days.Results. In vitro, the synovial fluid adherent cells consisted of 2 subpopulations, large round-shaped macrophage-like cells (CD68+) and spindle-shaped fibroblast-like cells (Thy-1+). When passaged, the latter cells proliferated and organized themselves into 3-dimensional formations. This allowed them to reach collagen particles fixed with agarose. Fibroblasts derived from synovial tissues could not be used in this assay because they grew only in monolayers and not on agarose. The majority (>90%) of passaged RA synovial fluid adherent cells expressed the Thy-1+,CD45-,CD68-,CD86- phenotype. Electron microscopy did not reveal important morphologic differences between the 2 types of fibroblasts, those from synovial tissue or those from synovial fluid. However, synovial fluid adherent cells expressed lower levels of adhesion molecules, including CD54 and galectin 3, as well as the complement-regulatory molecule CD55. The in vitro release of sGAG associated with cell activity was 2.5-fold higher from RA synovial fluid adherent cells in comparison with that from negative control cells. The release of sGAG correlated with the concentration of MMP-1 and was inhibited by the broad-range MMP inhibitor marimastat in a dose-dependent manner. RA synovial fluid adherent cells coimplanted with cartilage in SCID mice showed the same invasive behavior as that displayed by tissue-derived RA synovial fibroblasts.Conclusion. Similar to tissue-derived RA synovial fibroblasts, RA synovial fluid adherent cells, which contain "floating" anchorage-independent fibroblast-like cells, mediate cartilage destruction independent of the hyperplastic synovial tissue.