Characterization of fibroblasts responsible for cartilage destruction in arthritis

Abstract

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 aV), 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. '

Cyclophilin–CD147 interactions: a new target for anti-inflammatory therapeutics

“…For example, an up-regulated expression of CD147 was observed in the synovial membrane [132], as well as on circulating and synovial monocytes/macrophages [149], in RA patients. Increased levels of CD147 were also reported recently on a murine synovial fibroblast cell line used for studies of cartilage invasion and destruction [150]. One might postulate that an increase in CD147 expression would enhance the interaction between leucocytes and extracellular cyclophilins, thereby promoting their recruitment.…”

Au@pNIPAM SERRS Tags for Multiplex Immunophenotyping Cellular Receptors and Imaging Tumor Cells

“…Since these cell lines differed in the expression of the three selected biomarkers, this experiment was used to evaluate the targeting specificity of the SERRS tags. Whereas the A431 cell line expressed EGFR, EpCAM, and CD44, the 3T3 2.2 cell line expressed CD44, but neither EGFR nor EpCAM . To ascertain the differential expression of the biomarkers in A431 and 3T3 2.2 cell lines, immunofluorescence assays were also performed with the same specific antibodies used for Au@pNIPAM microgels bioconjugation.…”

NIH/3T3 Fibroblasts Selectively Activate T Cells Specific for Posttranslationally Modified Collagen Type II

“…Among the cell-cell contact-dependent interactions leading to T-cell activation, a role for CD154 (CD40L)-CD40 engagement has been highlighted. It has been shown that CD40 is expressed on synovial fibroblasts and destructive murine fibroblasts [ 80 , 81 ], and ligation of CD40 induces SF proliferation and up-regulation of adhesion molecules [ 68 , 69 ]. However, CD154 expression on T-cells is transient and is rapidly down-regulated after binding to CD40 [ 82 ].…”