Xu W D, Firestein G S, Taetle R, Kaushansky K, Zvaifler N J
Department of Medicine, University of California, San Diego 92103.
J Clin Invest. 1989 Mar;83(3):876-82. doi: 10.1172/JCI113971.
A liquid culture technique was used to study 23 synovial fluids (SF) (21 from inflammatory joint diseases and 2 noninflammatory SF) and supernatants of two cultured rheumatoid arthritis (RA) synovial tissues for colony-stimulating factor (CSF). The proliferative responses of human peripheral blood macrophage-depleted non-T cells treated with synovial fluids, supernatants of synovial tissue explants, and recombinant granulocyte-macrophage (rGM)-CSF were compared. Aggregates of cells that formed in long-term cultures (15 d) were similar for each applied agent and consisted of macrophages, eosinophils, and large blasts. Tritiated thymidine incorporation was proportional to the concentration of rGM-CSF and was accompanied by an increase in number and size of cellular aggregates formed in the cultures. CSF activity was observed in inflammatory SF, with tritiated thymidine uptake of 3,501 +/- 1,140 cpm in the presence of RA samples (n = 15) compared to 1,985 +/- 628 for non-RA inflammatory SF (n = 7) (P less than 0.05) and 583 +/- 525 for medium (n = 6) (P less than 0.01). The proliferative response to RA SF was often more apparent when the samples were diluted, because at higher concentrations the RA SF was inhibitory. Two RA SF were fractionated by Sephadex G100 column chromatography; low levels of CSF activity were detected in fractions corresponding to Mr of 70-100 kD, but the major CSF activity was found in the 20-24-kD fractions. A polyclonal rabbit anti-GM-CSF antibody eliminated the stimulating activity from both rGM-CSF and RA SF. Finally, a specific RIA identified significant levels of GM-CSF (40-140 U/ml) in the culture supernatants of 3 additional RA synovial tissues. These data document the local production of GM-CSF in rheumatoid synovitis and are the first description of this cytokine at a site of disease activity.
采用液体培养技术研究了23份滑液（SF）（21份来自炎性关节疾病，2份为非炎性滑液）以及两份培养的类风湿关节炎（RA）滑膜组织的上清液中的集落刺激因子（CSF）。比较了用滑液、滑膜组织外植体上清液和重组粒细胞-巨噬细胞（rGM）-CSF处理的人外周血去除巨噬细胞的非T细胞的增殖反应。长期培养（15天）中形成的细胞聚集体，对于每种应用试剂来说都相似，由巨噬细胞、嗜酸性粒细胞和大的母细胞组成。氚标记胸腺嘧啶核苷掺入量与rGM-CSF浓度成正比，并伴随着培养物中形成的细胞聚集体数量和大小的增加。在炎性滑液中观察到CSF活性，在存在RA样本时（n = 15），氚标记胸腺嘧啶核苷摄取量为3501±1140 cpm，而非RA炎性滑液（n = 7）为1985±628（P<0.05），培养基（n = 6）为583±525（P<0.01）。当样本稀释时，对RA滑液的增殖反应通常更明显，因为在较高浓度时RA滑液具有抑制作用。两份RA滑液通过Sephadex G100柱层析进行分离；在对应于分子量70 - 100 kD的级分中检测到低水平的CSF活性，但主要的CSF活性存在于20 - 24 kD的级分中。一种多克隆兔抗GM-CSF抗体消除了rGM-CSF和RA滑液的刺激活性。最后，一种特异性放射免疫分析（RIA）在另外3份RA滑膜组织的培养上清液中鉴定出显著水平的GM-CSF（浓度为40 - 140 U/ml）。这些数据证明了类风湿滑膜炎中GM-CSF的局部产生，并且是对该细胞因子在疾病活动部位的首次描述。
