Nevalainen T J, Märki F, Kortesuo P T, Grütter M G, Di Marco S, Schmitz A
Department of Pathology, University of Turku, Finland.
J Rheumatol. 1993 Feb;20(2):325-30.
Phospholipase A2 (PLA2) was produced in E. coli by a recombinant technique using a synthetic gene coding for PLA2 found in synovial fluid (SF) of patients suffering from rheumatoid arthritis (RA). Polyclonal antibodies were produced against the recombinant synovial type PLA2 (syn-PLA2) in a rabbit. The IgG fraction of the antiserum was isolated and the specificity was tested by immunoblotting. The antiserum detected a protein with an apparent molecular weight of 15 kDa in extracts of cartilage, but did not react with PLA2 isolated from human pancreas or ascitic fluid. In immunohistochemistry, the anti-syn-PLA2 antibody decorated chondrocytes and matrix of articular, laryngeal and auricular cartilage, but did not decorate synovial tissue or its contained inflammatory cells in RA or other human tissues tested (pancreas, liver, thyroid, tonsil, lung, nasal polyp, skin, placenta, myometrium, bone). The results show that syn-PLA2 is present both in articular and extraarticular cartilage and support the view that PLA2 found in SF might originate from chondrocytes, and not from synovial lining or inflammatory cells.
通过重组技术，利用编码类风湿性关节炎（RA）患者滑液（SF）中发现的磷脂酶A2（PLA2）的合成基因，在大肠杆菌中生产出磷脂酶A2。在兔体内产生了针对重组滑膜型PLA2（syn-PLA2）的多克隆抗体。分离出抗血清的IgG部分，并通过免疫印迹法检测其特异性。抗血清在软骨提取物中检测到一种表观分子量为15 kDa的蛋白质，但不与从人胰腺或腹水分离的PLA2发生反应。在免疫组织化学中，抗syn-PLA2抗体标记关节软骨、喉软骨和耳软骨的软骨细胞和基质，但不标记RA患者的滑膜组织或其中所含的炎症细胞，也不标记其他检测的人体组织（胰腺、肝脏、甲状腺、扁桃体、肺、鼻息肉、皮肤、胎盘、子宫肌层、骨骼）。结果表明，syn-PLA2存在于关节软骨和关节外软骨中，支持了SF中发现的PLA2可能起源于软骨细胞而非滑膜衬里或炎症细胞的观点。
