Characterization of fibroblasts responsible for cartilage destruction in arthritis.
In the pathogenesis of rheumatoid arthritis (RA), synovial fibroblasts (SF) play a key role as they secrete distinct patterns of cytokines and express variable levels of costimulatory and adhesion molecules. The murine fibroblast cell line LS48 has been shown to be invasive in the cartilage destruction models in vivo and in vitro. The purpose of this study was to examine in detail the LS48 phenotype, to obtain a better understanding of the SF-mediated cartilage destruction in RA. The destructive fibroblasts line LS48 and the nondestructive 3T3 cells were cultured and characterized with slide-based and flow cytometry, using antibodies against several adhesion molecules, immunological acting molecules, and marker proteins. The invasive LS48 fibroblasts are characterized by significantly higher expression of adhesion molecules such as CD47 (IAP), CD51 (integrin alpha V), CD61 (GPIIIa), and CD147 (EMMPRIN), and immunological acting molecules such as CD40 (Bp50), CD55 (DAF), and TLR-2. The results from the slide-based and flow cytometry analyses were exactly the same, except for the selected CD147 and TLR-2. This study demonstrated that the destructive fibroblast cell line LS48 has the characteristics of RA SFs. The high expression of specific costimulatory and adhesion molecules underlines the aberrant phenotype of these cells when compared with noninvasive fibroblasts. Furthermore, slide-based and flow cytometry complement each other in fibroblast phenotyping. Overall, this study shows that LS48 is an excellent tool to gain a deeper understanding of SF in RA.
Treese
C
C
Institute of Clinical Immunology and Transfusion Medicine, Medical Faculty, University of Leipzig, Leipzig, Germany.
Mittag
A
A
Lange
F
F
Tarnok
A
A
Loesche
A
A
Emmrich
F
F
Lehmann
J
J
Sack
U
U
D016475:3T3 Cells; D000818:Animals; D001168:Arthritis / Q000378:metabolism*; D002356:Cartilage / Q000378:metabolism*; D002448:Cell Adhesion; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D007158:Immunologic Techniques; D016130:Immunophenotyping / Q000379:methods*; D051379:Mice; D008954:Models, Biological; D010641:Phenotype; D013583:Synovial Membrane / Q000166:cytology