Summers K L, Daniel P B, O'Donnell J L, Hart D N
Haematology/Immunology Research Group, Christchurch Hospital, New Zealand.
Clin Exp Immunol. 1995 Apr;100(1):81-9. doi: 10.1111/j.1365-2249.1995.tb03607.x.
Dendritic cells (DC) act as potent primary antigen-presenting cells in many immune responses and therefore may have a role in the initiation and perpetuation of the synovial inflammation in chronic inflammatory arthritis. To examine their function, it is important to isolate fresh DC from arthritic joints without aberrant activation. We have developed a technique using minimal cell manipulation to isolate DC from the synovial fluid of chronic arthritic patients. Using this method, DC were shown to be potent allostimulatory cells, with 63-90% of cells lacking lineage-specific markers (lin-), but positive for MHC class II molecules. Two morphologically distinct populations of these cells were identified in 10 out of 13 DC preparations. Both populations expressed CD40, intercellular adhesion molecule-1 (ICAM-1), ICAM-2, ICAM-3 and leucocyte function associated antigen-3 (LFA-3), but the predominant population, which was larger and more typical of cultured blood DC, had a higher density of these antigens compared with the minor population, which were smaller and morphologically similar to lymphocytes. Two new MoAbs which label activated human blood DC, HB15 (CD83) and CMRF-44, were tested. CD83 labelled very weakly or not at all, whereas CMRF-44 was positive on the larger cells only. Likewise, the costimulator molecule, B7/BB1 (CD80), was not detected on the surface of either synovial lin- cell population, reverse transcriptase polymerase chain reaction (RT-PCR) showed little or no CD80 mRNA, and no binding of the CTLA-4Ig fusion protein was found. These results suggest that synovial DC are not, despite the inflammatory environment, in a fully activated state.
树突状细胞(DC)在许多免疫反应中作为强大的主要抗原呈递细胞,因此可能在慢性炎症性关节炎的滑膜炎症的起始和持续中发挥作用。为了研究它们的功能,重要的是从关节炎关节中分离出未发生异常激活的新鲜DC。我们开发了一种使用最少细胞操作的技术,从慢性关节炎患者的滑液中分离DC。使用这种方法,DC被证明是强大的同种异体刺激细胞,63 - 90%的细胞缺乏谱系特异性标志物(lin-),但MHC II类分子呈阳性。在13份DC制剂中的10份中鉴定出了两种形态不同的细胞群体。这两种群体均表达CD40、细胞间黏附分子-1(ICAM-1)、ICAM-2、ICAM-3和白细胞功能相关抗原-3(LFA-3),但占主要的群体,其细胞较大且更具培养血液DC的典型特征,与较小且形态与淋巴细胞相似的次要群体相比,这些抗原的密度更高。测试了两种标记活化人血液DC的新单克隆抗体HB15(CD83)和CMRF-44。CD83标记非常弱或根本不标记,而CMRF-44仅在较大细胞上呈阳性。同样,共刺激分子B7/BB1(CD80)在滑膜lin-细胞群体的表面均未检测到,逆转录聚合酶链反应(RT-PCR)显示几乎没有或没有CD80 mRNA,并且未发现CTLA-4Ig融合蛋白的结合。这些结果表明,尽管存在炎症环境,但滑膜DC并未处于完全激活状态。