Immunolocalization of type X collagen in normal fetal and adult osteoarthritic cartilage with monoclonal antibodies

Matrix Biol. 1996 Sep;15(4):231-8. doi: 10.1016/s0945-053x(96)90114-6.


For studies on processing and tissue distribution of type X collagen, monoclonal antibodies were prepared against human recombinant collagen type X (hrCol X) and tested by ELISA, immunoblotting and immunohistology. Forty-two clones were obtained which were grouped into four different subsets based on their reactivity against native and denatured hrCol X, pepsin-treated hrCol X, and the C-terminal NC-1 domain. Here we present results obtained with four monoclonal antibodies: Clone X 53, a representative of group I, binds with high affinity to both native and pepsin-digested hrCol X but with low affinity to the NC-1 dimer; monoclonal antibodies of group II and III recognized native and denatured hrCol X but not NC-1; antibodies of group II, but not III, reacted to some extent with pepsin treated hrCol X; one antibody (X 34) was obtained that reacted strongly with the isolated NC-1 dimer and native hrCol X but not with the NC-1 monomer or pepsin-digested hrCol X (group IV). Antibodies of all groups stained specifically the hypertrophic zone of fetal human epiphyseal cartilage. Mab X 53 stained the peri- and extracellular matrix of hypertrophic chondrocytes in the lower hypertrophic zone and in the calcified cartilage core in endochondral bone trabecules, while clone X 34 stained intracellularly and the pericellular matrix. All other tissues or cells of the epiphysis were negative. Antibody X 53 reacted also with canine, murine and guinea pig hypertrophic cartilage in tissue sections, but not with bovine or porcine type X collagen. In sections of osteoarthritic cartilage, clusters of hypertrophic chondrocytes in the deep zone were stained, confirming previous observations on enhanced chondrocyte hypertrophy and type X collagen expression in osteoarthritic articular cartilage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Antibodies, Monoclonal
  • Antibody Specificity
  • Cartilage, Articular / cytology*
  • Cartilage, Articular / pathology*
  • Cattle
  • Collagen / analysis*
  • Collagen / immunology
  • Dogs
  • Fetal Death
  • Fetus
  • Gestational Age
  • Humans
  • Hypertrophy
  • Immunoblotting
  • Immunohistochemistry
  • Mice
  • Mice, Inbred BALB C
  • Osteoarthritis / pathology*
  • Protein Denaturation
  • Recombinant Proteins / immunology
  • Swine


  • Antibodies, Monoclonal
  • Recombinant Proteins
  • Collagen