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. 1997 Dec;56(12):729–736. doi: 10.1136/ard.56.12.729

Differential expression and functional behaviour of the αv and β3 integrin subunits in cytokine stimulated fibroblast-like cells derived from synovial tissue of rheumatoid arthritis and osteoarthritis in vitro

N Rinaldi 1, D Weis 1, B Brado 1, M Schwarz-Eywill 1, M Lukoschek 1, A Pezzutto 1, U Keilholz 1, T Barth 1
PMCID: PMC1752301  PMID: 9496152

Abstract

OBJECTIVE—The aim of this study was to investigate in situ the expression of the classic vitronectin (VN) receptor consisting of the αv and β3 subunits in synovial lining cells (SLC) of chronic synovitis occurring in osteoarthritis (OA) and in rheumatoid arthritis (RA). The expression and function of αv and β3 as VN receptor in cultured fibroblast-like synoviocytes (FBS) derived from patients with OA and RA was also compared.
METHODS—Expression of αv and β3 was examined immunohistochemically in normal synovial tissue and in synovial tisssue from patients with OA and RA. The effect of proinflammatory cytokines and of a synovial fluid of a patient with RA on the expression of the αv and β3 subunits of cultured FBS was determined by flow cytometry. Binding of OA and RA-FBS to VN was quantified using adhesion assays and the effect of interleukin 1β (IL1β) and tumour necrosis factor α (TNFα) on adhesion was measured. The specifity of the adhesion was tested by inhibition studies using monoclonal antibodies to integrin subunits.
RESULTS—In in situ studies normal SLC showed a parallel distribution of αv and β3 subunits. OA-SLC strongly and uniformly expressed αv whereas RA-SLC showed heterogeneous expression of αv. In situ both OA-SLC and RA-SLC lacked the expression of the integrin subunit β3. In in vitro studies, OA-FBS and RA-FBS did not differ as regards expression of αv and β3, and VN attachment. Binding of RA-FBS to VN was partially blocked by antibodies against αv, β1, and β3 subunits, whereas only antibodies against αv and β3 inhibited the binding of OA-FBS to VN. The proinflammatory cytokines TNFα and IL1β increased the expression of αv and β3, and the VN binding of OA-FBS, whereas αv and β3 expression, and VN binding were downregulated in RA-FBS. Similar effects were found when the synovial fluid of an RA patient was used.
CONCLUSION—The integrin subunit β3 seems to be one partner but not the major one with which the subunit αv forms functional vitronectin receptors in OA-FBS and RA-FBS. The interaction between synovial cells and inflammatory cytokines seems to be different for OA and RA; the basis for this difference, however, remains to be established.



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Figure 2  .

Figure 2  

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Histograms of flow cytometric analysis for the αv integrin subunit on FBS from OA (upper panels) and from RA (lower panels). The fluorescence obtained with isotypic negative control antibodies is plotted as a plain line, the results with anti-αv as a bold line.

Figure 3  .

Figure 3  

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Effect of cytokines and synovial fluid (SF) from an arthritic joint of a RA patient on the binding of FBS cultures derived from two OA patients and from two RA patients, as determined by adhesion assay.

Figure 4  .

Figure 4  

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Inhibition of adhesion of FBS from OA and RA to VN. Monoclonal antibodies (mAb) against α3, α5, α6, αv, β1, and β3 were used; however only mAbs that blocked FBS binding are shown in the figure. Inhibition through the mAb is given as relative per cent adhesion compared with adhesion without antibodies (100%). Data represent the means of assays of three OA-FBS cultures and of three RA-FBS cultures.

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