Effects of TIMP-1 and TIMP-3 gene transfer on invasiveness, proliferation and apoptosis of rheumatoid arthritis (RA) synovial fibroblasts (RA-SF)

  • T Pap1,

    Affiliated with

    • A Drynda1,

      Affiliated with

      • CA Seemayer2,

        Affiliated with

        • PHA Quax3,

          Affiliated with

          • JH Verheijen3,

            Affiliated with

            • S Drynda1,

              Affiliated with

              • TWJ Huizinga4,

                Affiliated with

                • BA Michel5,

                  Affiliated with

                  • RE Gay2,

                    Affiliated with

                    • S Gay2 and

                      Affiliated with

                      • WH van der Laan6

                        Affiliated with

                        Arthritis Res20024(Suppl 1):109

                        DOI: 10.1186/ar444

                        Received: 15 January 2002

                        Published: 4 February 2002

                        TIMPs play a key role in counter balancing the action of MMPs and have been associated with cell proliferation, inhibition of angiogenesis and induction of apoptosis. Here, we investigated the effects of TIMP-1 and TIMP-3 gene transfer on cartilage invasion, proliferation and apoptosis of RA-SF. RA-SF were transduced with an adenoviral vector expressing human TIMP-1 (AdTIMP-1) or TIMP-3 (AdTIMP-3). Transduction efficacy was assessed by LacZ staining of RA-SF that were transduced with an adenoviral β-galactosidase construct. Untransduced and mock transduced RA-SF were used as controls. TIMP-1 was measured by ELISA in the culture supernatants of AdTIMP-1 transduced and mock transduced cells every 10 days until 60 days after transduction. Proliferation was assessed by 3H-thymidine incorporation, and the rate of spontaneous apoptosis as well as FasL induced cell death was determined by a histon fragmentation assay. AdTIMP-1 and AdTIMP-3 transduced RA-SF and control RA-SF were co-implanted with human articular cartilage under the renal capsule of SCID mice for 60 days and their invasiveness was evaluated on paraffin sections using a semiquantitative score. Transduction efficacy was 67%, and TIMP-1 levels in the supernatants of AdTIMP-1 transduced cells were 51.5 ± 6.5 μg/ml as compared to 8.7 ± 3.4 μg/ml in the mock transduced cells. These levels of TIMP expression were maintained for at least 60 days. AdTIMP-1 and AdTIMP-3 gene transfer resulted in an inhibition of proliferation (35% and 40% vs. mock, respectively; P < 0.05). Transduction of RA-SF with AdTIMP-3 but not TIMP-1 increased spontaneous apoptosis (+24%; vs. mock, P < 0.05) as well as susceptibility to FasL-induced cell death (+23% vs. mock, P < 0.05). In the SCID mouse model, untransduced and mock transduced RA.SF deeply invaded the cartilage (scores: 2.5 ± 0.2 and 3.2 respectively). In the AdTIMP-1 and AdTIMP-3 transduced RA-SF, invasion was inhibited clearly (scores 0.9 ± 0.4 and 1.2 ± 0.2 respectively) Both AdTIMP-1 and AdTIMP-3 gene transfer inhibit proliferation of RA-SF and reduce cartilage invasion. In contrast to TIMP-1, adenoviral gene transfer with TIMP-3. has a strong pro-apoptotic effect on RA-SF and facilitates Fas mediated cell death. These results indicate that gene transfer of TIMPs may be a useful approach to inhibit joint destruction in RA.

                        Authors’ Affiliations

                        (1)
                        Division of Experimental Rheumatology
                        (2)
                        Center of Exp Rheumatology
                        (3)
                        TNO Prevention and Health
                        (4)
                        Dept of Rheumatology
                        (5)
                        Dept. Rheumatology
                        (6)
                        TNO Prevention and Health

                        Copyright

                        © BioMed Central Ltd 2002

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