IMMUNOHISTOCHEMICAL DETECTION AND IMMUNOCHEMICAL ANALYSIS OF TYPE-II COLLAGEN DEGRADATION IN HUMAN NORMAL, RHEUMATOID, AND OSTEOARTHRITIC ARTICULAR CARTILAGES AND IN EXPLANTS OF BOVINE ARTICULAR-CARTILAGE CULTURED WITH INTERLEUKIN-1
IMMUNOHISTOCHEMICAL DETECTION AND IMMUNOCHEMICAL ANALYSIS OF TYPE-II COLLAGEN DEGRADATION IN HUMAN NORMAL, RHEUMATOID, AND OSTEOARTHRITIC ARTICULAR CARTILAGES AND IN EXPLANTS OF BOVINE ARTICULAR-CARTILAGE CULTURED WITH INTERLEUKIN-1
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DOI：
10.1172/jci113929
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发表时间：
1989-02-01
影响因子：
15.9
通讯作者：
POOLE, AR
中科院分区：
文献类型：
--
作者：
DODGE, GR;POOLE, AR
关节炎性疾病中关节软骨的破坏和功能丧失涉及结缔组织基质的蛋白水解降解。我们已经研究了软骨胶原的降解，通过发展免疫化学方法，允许识别和分析II型胶原原位降解。以前，一种技术，以具体确定II型胶原降解原位关节软骨不存在。这些方法利用多克隆抗血清（R181），其特异性地与解绕的α-人和牛II型胶原蛋白的α 1（II）CB 11和α 1（II）CB 8链和CNBr衍生肽。实验方法是基于这样的事实，即当纤维状胶原蛋白被切割时，螺旋胶原蛋白分子解旋，暴露隐藏的表位。在这里，我们证明了使用R181在研究II型胶原降解牛关节软骨，已培养或不IL-1和人类正常，类风湿性关节炎和骨关节炎关节软骨。与新鲜分离的软骨或无IL-1培养的软骨相比，IL-1培养3-5 d的牛软骨细胞周和细胞间免疫组化染色均增加。这些软骨的提取物含有II型胶原蛋白α。IL-1作用11 d后，IL-1 mRNA表达量增加。此外，用IL-1培养导致α的出现。较低分子量的链片段。所有人类关节炎组织检查显示，与非病变组织相比，明显的细胞周围和领土染色的胶原蛋白降解的区域，表明软骨细胞是负责这种降解的部分与非病变组织相比。在大多数情况下，类风湿性关节炎软骨在关节面和深中间区染色最强烈，而骨关节炎软骨通常在浅表和中间区染色较多，但强度较低。胶原降解位点的不同模式反映了这些疾病中胶原破坏的差异，表明软骨细胞活化可能有不同的来源。这些实验证明了免疫学方法在检测胶原降解中的应用，并证明了人关节炎和IL-1处理的活牛软骨中胶原降解的增加。
Articular cartilage destruction and loss of function in arthritic diseases involves proteolytic degradation of the connective tissue matrix. We have investigated the degradation of cartilage collagen by developing immunochemical methods that permit the identification and analysis of type II collagen degradation in situ. Previously, a technique to specifically identify type II collagen degradation in situ in articular cartilage did not exist. These methods utilize a polyclonal antiserum (R181) that specifically reacts with unwound .alpha.-chains and CNBr-derived peptides, .alpha.1(II)CB11 and .alpha.1(II)CB8, of human and bovine type II collagens. The experimental approach is based on the fact that when fibrillar collagens are cleaved the helical collagen molecular unwinds, exposing hidden epitopes. Here we demonstrate the use of R181 in studying type II collagen degradation in bovine articular cartilage that has been cultured with or without IL-1 and in human normal, rheumatoid, and osteoarthritic articular cartilages. Compared to cartilages either freshly isolated or cultured without IL-1, bovine cartilage cultured with IL-1 for 3-5 d showed an increase in both pericellular and intercellular immunohistochemical staining. Extracts of these cartilages contained type II collagen .alpha. chains that were increased in amount after culture with IL-1 for 11 d. In addition, culture with IL-1 resulted in the appearance of .alpha. chain fragments of lower molecular weight. All human arthritic tissues examined showed areas of pronounced pericellular and territorial staining for collagen degradation as compared with nondiseased tissues, indicating that chondrocytes are responsible in part for this degradation as compared with nondiseased tissues. In most cases rheumatoid cartilage was stained most intensely at the articular surface and in the deep and midzones, whereas osteoarthritic cartilage usually stained more in the superficial and mid-zones, but less intensely. Distinct patterns of sites of collagen degradation reflect differences in collagen destruction in these diseases, suggesting possible different sources of chondrocyte activation. These experiments demonstrate the application of immunological methods to detect collagen degradation and demonstrate an increase of collagen degradation in human arthritides and in IL-1-treated viable bovine cartilage.