The Activating Effect of Synovial Fluid and Washings of Synovial Membrane on Autologous Lymphocytes in Rheumatoid Arthritis

Abstract

The effect of synovial fluid and washings of synovial membrane on autologous lymphocytes from patients with rheumatoid arthritis and other diseases has been studied using a rapid method based upon the increase in intranuclear birefringence occurring in the early stages of lymphocyte activation. Retardation of polarized light indicating increased lymphocyte activation was seen in lymphocytes from patients with rheumatoid arthritis but not in lymphocytes from patients with other diseases.Recent studies have shown that, when examined by electron microscopy, lymphocytes present in the synovial membrane of patients suffering from rheumatoid arthritis present the appearances of well developed blastic cells or the intermediate states of blastic transformation (l,2). The increased uptake of radioactive thymidine by isolated synovial fluid lymphocytes (3) that not only the lymphocytes in the rheumatoid synovial membrane but also those in the synovial fluid are being activated. It has been reported that while the lymphocytes of the synovial fluid are being activated, the blood lymphocytes of the same patient show characteristics within normal limits (3). Therefore it appears likely that the lymphocytes become activated within the synovial membrane (4) or within the synovial effusion as a result of a factor or factors present there.Isolated blood lymphocytes of patients suffering from rheumatoid arthritis and other diseases were examined in vitro to see whether they would become activated when added to a cell-free synovial effusion or washings from synovial membrane obtained from the same patient, T o detect the activated lymphocytes, a new rapid method was used based upon nuclear birefringence occurring lymphocyte activation ( 5 ) . MATERIALS ANDFifteen milliliters of blood the increase in intrain the early stages of METHODSwere obtained by venipuncture and mixed with 1000 IU of heparin. The lymphocytes were isolated by phycol-isopaque gradient centrifugation described by Boyum (6). T h e isolated lymphocytes were cultured in TC 199 (Difco) medium. The number of the living mononuclear cells was between 0.9 and 1.5 X IOB in each milliliter of the culture medium. T h e culture medium was divided into samples (each 2 ml in volume). One sample was used a s a control while others were treated with 0.1; 0.2; 0.4 ml cell-free supernatant of the centrifuged synovial effusion.

Membrane and transformation characteristics of lymphocytes isolated from the synovial membrane and paired peripheral blood of patients with rheumatoid arthritis

“…The higher spontaneous 3Hthymidine uptake of synovial tissue lymphocytes observed at days 3 and 6 as compared to peripheral blood lymphocytes favours this explanation. The activating effect of synovial fluid on autologous peripheral blood lymphocytes in rheumatoid arthritis (Sebok et al, 1977) is also compatible with this explanation.…”