Gulati P, Guc D, Lemercier C, Lappin D, Whaley K
Department of Immunology, Leicester Royal Infirmary, UK.
Rheumatol Int. 1994;14(1):13-9. doi: 10.1007/BF00302666.
We studied the synthesis of the classical pathway complement components in synovial membrane. Ribonucleic acid was extracted from the synovial membranes of patients with rheumatoid arthritis (RA) or osteoarthritis (OA), as well as from normal synovial membrane. Northern blot and dot blot analysis showed that the mRNAs for all classical pathway complement components (C1qA chain, C1qB chain, C1qC chain, C1r, C1s, C4 and C2) and the fluid-phase regulatory components (C1-inhibitor, C4-bp and factor I) were present in all three types of synovial membrane. Thus, all the components of the classical pathway were expressed in normal and diseased synovium. In an attempt to determine which components were synthesised by each cell type, monocytes (mononuclear phagocytes), human umbilical vein endothelial cells (HUVEC), synovial membrane fibroblasts (from normal, OA and RA synovial membrane) and peripheral blood lymphocytes were cultured in vitro and secretion rates of individual components were measured and total cellular RNA was analysed by Northern blotting. Monocytes secreted C1q, C1r, C1s, C4, C2, C1-inhibitor and C4-bp but not factor I. Fibroblasts secreted C1r, C1s, C2, C3, C1-inhibitor and factor I but not C1q, C4 or C4-bp. HUVEC secreted C1s, C2, C1-inhibitor and factor I but not C1q, C1r, C4 or C4-bp. Lymphocytes did not secrete any of these components. In three instances mRNA was detected in the absence of secreted protein: mRNAs for the C1qA and C1qC chains were detected in HUVEC, whereas the mRNA for the C1qB chain was not, and C4 mRNA was detected in both fibroblasts and HUVEC.(ABSTRACT TRUNCATED AT 250 WORDS)
我们研究了滑膜中经典途径补体成分的合成。从类风湿性关节炎(RA)或骨关节炎(OA)患者的滑膜以及正常滑膜中提取核糖核酸。Northern印迹和斑点印迹分析表明,所有经典途径补体成分(C1qA链、C1qB链、C1qC链、C1r、C1s、C4和C2)以及液相调节成分(C1抑制因子、C4结合蛋白和因子I)的信使核糖核酸在所有三种类型的滑膜中均有存在。因此,经典途径的所有成分在正常和患病滑膜中均有表达。为了确定每种细胞类型合成哪些成分,将单核细胞(单核吞噬细胞)、人脐静脉内皮细胞(HUVEC)、滑膜成纤维细胞(来自正常、OA和RA滑膜)和外周血淋巴细胞进行体外培养,测量各成分的分泌率,并通过Northern印迹分析总细胞核糖核酸。单核细胞分泌C1q、C1r、C1s、C4、C2、C1抑制因子和C4结合蛋白,但不分泌因子I。成纤维细胞分泌C1r、C1s、C2、C3、C1抑制因子和因子I,但不分泌C1q.C4或C4结合蛋白。HUVEC分泌C1s、C2、C1抑制因子和因子I,但不分泌C1q、C1r、C4或C4结合蛋白。淋巴细胞不分泌这些成分中的任何一种。在三个实例中,在没有分泌蛋白的情况下检测到了信使核糖核酸:在HUVEC中检测到了C1qA和C1qC链的信使核糖核酸,而未检测到C1qB链的信使核糖核酸,并且在成纤维细胞和HUVEC中均检测到了C4信使核糖核酸。(摘要截短于250字)