Osteoarthritis in cynomolgus macaques. II. Detection of modulated proteoglycan epitopes in cartilage and synovial fluid.
Osteoarthritis in cynomolgus macaques. II. Detection of modulated proteoglycan epitopes in cartilage and synovial fluid.
复制标题
食蟹猴的骨关节炎。
DOI：
10.1002/jor.1100130314
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Caterson,B
中科院分区：
文献类型：
--
作者：
Carlson,CS;Loeser,RF;Johnstone,B;Tulli,HM;Dobson,DB;Caterson,B
本研究的目的是确定单克隆抗体7-D-4和3-B-3作为成年食蟹猴膝关节中自然发生的骨关节炎严重程度的生物标志物的有用性。抗体用于免疫定位关节软骨或关节滑液中的硫酸软骨素蛋白聚糖表位，所述关节软骨或关节滑液来自具有一系列骨关节炎严重程度的膝关节。对关节进行放射学、大体、显微放射学和组织学检查，以表征疾病的严重程度，并比较三种不同的蛋白多糖分析方法（免疫组织化学、酶联免疫吸附试验和蛋白质印迹分析）的结果。主观上，7-D-4的阳性免疫染色程度在正常部位是最小的，并且随着关节软骨损伤的增加而增加。胫骨平台内侧（该模型中受累最严重的部位）7-D-4免疫染色评分与关节软骨损伤的严重程度显著相关（p < 0.05，r2= 0.50），因此支持主观观察结果。滑液中7-D-4与硫酸化糖胺聚糖的比值也与胫骨平台内侧7-D-4免疫染色评分（p < 0.05，r2= 0.54）和股骨髁内侧3-B-3免疫染色评分（p < 0.05，r2= 0.65）相关。3-B-3免疫染色评分、严重程度评分和滑液中3-B-3与硫酸化糖胺聚糖的比值之间无显著相关性。通过Western印迹分析，这两个表位是年轻成年猴早期软骨损伤的敏感标志物，但在老年猴中不太敏感。这项工作提供的证据表明，测量7-D-4在滑液中识别的表位，或通过免疫组织化学或蛋白质印迹方法在关节软骨中识别的表位，有可能用作自然发生的骨关节炎严重程度的标志物。
The purpose of the present study was to determine the usefulness of the monoclonal antibodies 7‐D‐4 and 3‐B‐3 as biomarkers of severity of naturally occurring osteoarthritis in the knee joints of adult cynomolgus macaques. The antibodies were used to immunolocate chondroitin sulfate proteoglycan epitopes in articular cartilage or synovial fluid from knee joints with a range in severity of osteoarthritis. The joints were examined radiographically, grossly, microradiographically, and histologically to characterize the severity of disease, and the results of three different methods of proteoglycan analysis (immunohistochemistry, enzyme‐linked immunosorbent assay, and Western blot analysis) were compared. Subjectively, the degree of positive immunostaining for 7‐D‐4 was minimal in normal sites and increased as damage to articular cartilage increased. The scores for 7‐D‐4 immunostaining in the medial tibial plateau (the site most severely involved in this model) were correlated significantly with severity of damage to articular cartilage (p < 0.05, r2= 0.50), thus supporting the subjective observations. The ratio of 7‐D‐4 to sulfated glycosaminoglycans in synovial fluid also was correlated with the score for 7‐D‐4 immunostaining in the medial tibial plateau (p < 0.05, r2= 0.54) and with the score for 3‐B‐3 immunostaining in the medial femoral condyle (p < 0.05, r2= 0.65). There were no significant correlations among scores for 3‐B‐3 immunostaining, severity scores, and the ratios of 3‐B‐3 to sulfated glycosaminoglycans in the synovial fluid. By Western blot analysis, both epitopes were sensitive markers of early cartilage damage in young adult monkeys but were less sensitive in older monkeys. This work provides evidence that measurement of the epitope recognized by 7‐D‐4 in synovial fluid or, by immunohistochemical or Western blot methods, in articular cartilage has potential use as a marker of severity of naturally occurring osteoarthritis.