Immunolocalization of type X collagen in normal fetal and adult osteoarthritic cartilage with monoclonal antibodies.
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.
Institute of Experimental Medicine, Friedrich-Alexander-University of Erlangen-Nuremberg, Germany.
I
I
Girkontaite
S
S
Frischholz
P
P
Lammi
K
K
Wagner
B
B
Swoboda
T
T
Aigner
K
K
Von der Mark
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002358:Cartilage, Articular / Q000166:cytology* / Q000473:pathology*; D002417:Cattle; D003094:Collagen / Q000032:analysis* / Q000276:immunology; D004285:Dogs; D005313:Fetal Death; D005333:Fetus; D005865:Gestational Age; D006801:Humans; D006984:Hypertrophy; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010003:Osteoarthritis / Q000473:pathology*; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000276:immunology; D013552:Swine
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D003094:Collagen