Visualization of Type I and II Collagens in Tissue Sections by Immunohistologic Techniques

Abstract

Rabbit and rat antibodies were prepared against Type I and II collagens derived from bovine skin and articular cartilage, respectively. As judged by passive hemagglutination and radioimmune assays, these antibodies could be rendered generally specific for the type of collagen used for immunization by immunoadsorption. Thus, antibodies to Type II collagen did not cross-react with Type I and III collagens from skin. However, antibodies to Type I collagen still showed some cross-reaction with Type III collagen. Antibodies to Type I procollagen showed a negligible degree of cross-reaction with Type III procollagen. These purified antibodies reacted strongly with bovine and human tissue collagen as demonstrated by indirect immunoflourescence. Antibodies to Type I collagen stained dermal tissue, perichondral tissue, kidney stroma, aortic tissue, and annulus fibrosus. Antibodies to Type II collagen stained mainly the hyaline matrix of rib cartilage and nucleus pulposus. The staining patterns with anti-Type I procollagen were similar but not identical to that found with antibodies to Type I collagen. Neither of these antibodies reacted with kidney glomerular basement membrane. These antibody reagents are recommended as a sensitive and rapid screening tool for studying tissue distribution of collagen under normal and pathologic conditions.