Firestein G S, Xu W D, Townsend K, Broide D, Alvaro-Gracia J, Glasebrook A, Zvaifler N J
Department of Medicine, University of California, San Diego 92103.
J Exp Med. 1988 Nov 1;168(5):1573-86. doi: 10.1084/jem.168.5.1573.
Because previous studies showed low levels of IFN-gamma in rheumatoid arthritis (RA) synovial fluid (SF) and synovial tissue (ST) explant supernatants, we assayed RA SF and ST for IL-2 and IL-3-like activity. Using an IL-2 dependent murine CTLL line, 6 of 14 RA SF caused increased thymidine uptake (greater than three times control). The activity was distinct from IL-2 because it was not blocked by antibody to IL-2-R. In addition, IL-2 was not detected (less than 50 pg/ml) in 16 joint samples using an ELISA. Multi-colony-stimulating factor (CSF) activity was measured using two assays that can detect murine IL-3 (mast cell proliferation, and bone marrow CSF). In the mast cell assay, [3H]TdR uptake was 493 +/- 67 cpm for medium, 2,910 +/- 329 cpm in the presence of RA SF (p less than 0.001), 1,246 +/- 156 cpm in the presence of SF from patients with seronegative spondyloarthropathies (p less than 0.001), and 736 +/- 100 cpm in the presence of osteoarthritis SF (p greater than 0.1). In the CSF assay, four of five RA SF and five of five RA ST induced colony formation from bone marrow nonadherent cells. Macrophage colonies were most common, although mixed colonies and granulocytes were occasionally observed. The multi-CSF activity in RA is not due to IL-3 since human rIL-3 was not active in either murine assay, and IL-3 mRNA was not detected in RA synovium. Sephadex column chromatography of RA SF revealed that the mast cell growth factor (approximately 6 x 10(3) mol wt) and the CSF (approximately 40 and 100 x 10(3) mol wt) are distinct. The colony-stimulating aspect of the "IL-3-like" activity in RA SF is likely due to CSF-1 because it is the appropriate mol wt and because the activity was neutralized by specific anti-CSF-1 antibody. Finally, an RIA detected 1.6-25 ng/ml of CSF-1 in RA SF and ST and CSF-1 mRNA was detected in four of five RA synovial tissue samples tested.
因为先前的研究显示类风湿性关节炎（RA）滑液（SF）和滑膜组织（ST）外植体上清液中的γ干扰素水平较低，所以我们检测了RA SF和ST中的白细胞介素-2（IL-2）及白细胞介素-3样活性。使用依赖IL-2的小鼠CTLL细胞系，14份RA SF中的6份导致胸苷摄取增加（大于对照的三倍）。该活性与IL-2不同，因为它不被抗IL-2-R抗体阻断。此外，使用酶联免疫吸附测定法（ELISA）在16份关节样本中未检测到IL-2（小于50 pg/ml）。使用两种可检测小鼠IL-3的测定法（肥大细胞增殖和骨髓集落刺激因子（CSF））测量多集落刺激因子（CSF）活性。在肥大细胞测定中，培养基的[3H]TdR摄取为493±67 cpm，在RA SF存在下为2,910±329 cpm（p＜0.001），在血清阴性脊柱关节病患者的SF存在下为1,246±156 cpm（p＜0.001），在骨关节炎SF存在下为736±100 cpm（p＞0.1）。在CSF测定中，5份RA SF中的4份和5份RA ST中的5份诱导骨髓非贴壁细胞形成集落。巨噬细胞集落最常见，不过偶尔也观察到混合集落和粒细胞。RA中的多CSF活性并非由于IL-3，因为重组人IL-3在两种小鼠测定中均无活性，且在RA滑膜中未检测到IL-3信使核糖核酸（mRNA）。对RA SF进行葡聚糖凝胶柱色谱分析显示，肥大细胞生长因子（约6×10³分子量）和CSF（约40和100×10³分子量）是不同的。RA SF中“IL-3样”活性的集落刺激方面可能归因于CSF-1，因为其分子量合适，且该活性被特异性抗CSF-1抗体中和。最后，放射免疫测定法在RA SF和ST中检测到1.6 - 25 ng/ml的CSF-1，并且在所检测的5份RA滑膜组织样本中有4份检测到CSF-1 mRNA。
