Integrin expression by primary and immortalized human chondrocytes: evidence of a differential role for alpha1beta1 and alpha2beta1 integrins in mediating chondrocyte adhesion to types II and VI collagen.
OBJECTIVE
Chondrocytes have been shown to express beta1-containing integrins both in vitro and in situ, but their role in regulating chondrocyte function is poorly understood. The objective of this study was to determine how the relative expression of different integrins may be modulated in relation to the differentiated state and proliferative capacity of the chondrocyte.


METHODS
Integrin expression by four different cell lines of human chondrocytes immortalized with Simian virus 40 large T-antigen (SV40-TAg) was studied and compared to primary chondrocytes. Differences in alpha1 and alpha2 integrin subunit expression were utilized to further study the role of these integrins in mediating adhesion to types II and VI collagen.


RESULTS
The overall cell-surface levels of beta1-containing integrins were higher on all four immortalized cell lines which expressed over 10-fold higher levels of alpha2 and alpha3 integrin subunits compared to primary cells. However, primary cells expressed higher levels of the alpha1 integrin subunit which was not expressed by T/C28a4 cells and expressed at variable and lower levels in the other lines. Levels of the alpha3 integrin subunit were significantly greater on the highly proliferative juvenile costal chondrocyte lines (T/C-28a4, C-2812, and C-20a4) compared to primary articular chondrocytes and tsT/AC-62 cells which were derived from adult articular chondrocytes. Expression of alpha5 was similar among primary cells and cell lines except on C-20/A4 cells which had an average of over 4-fold higher levels. None of the primary or immortalized chondrocytes tested expressed significant levels of alpha4. Cell adhesion assays revealed that both alpha1beta1 and alpha2beta1 could serve as chondrocyte adhesion receptors for types II and VI collagen. In cell lines expressing both integrins, alpha1beta1 was the preferential receptor for type VI collagen while alpha2beta1 was the preferential receptor for type II collagen. Rather than inhibiting adhesion, incubation with the alpha3 blocking antibody P1B5 increased adhesion of C-28/12 cells to both fibronectin and type II collagen by 67% and 100% respectively.


CONCLUSIONS
Immortalization with SV40-TAg results in altered integrin expression by chondrocytes. Changes in the relative levels of alpha1, alpha2, and alpha3 subunits may significantly alter the manner in which chondrocytes interact with types II and VI collagen in the extracellular matrix.
Loeser
R F
RF
Department of Internal Medicine, The Wake Forest University School of Medicine, Winston-Salem, NC, USA. rloeser@rush.edu
Sadiev
S
S
Tan
L
L
Goldring
M B
MB
D000328:Adult; D000704:Analysis of Variance; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002461:Cell Line, Transformed / Q000378:metabolism; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D003094:Collagen / Q000502:physiology*; D005353:Fibronectins / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D006801:Humans; D039222:Integrin alpha1beta1; D016023:Integrins / Q000235:genetics* / Q000276:immunology; D008858:Microscopy, Phase-Contrast; D012333:RNA, Messenger / Q000235:genetics; D038981:Receptors, Collagen; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D039222:Integrin alpha1beta1; D016023:Integrins; D012333:RNA, Messenger; D038981:Receptors, Collagen; D003094:Collagen