Cytokine production in whole blood cell cultures of patients with rheumatoid arthritis

Abstract

Objective-The measurement of cytokine production of activated lymphocytes and monocytes in the whole blood cell (WBC) culture system may provide a sensitive tool for evaluating the actual ongoing immune response of patients with rheumatoid arthritis (RA). Methods-Lipopolysaccharide (LPS) up to 250 pg/ml was used for the stimulation of monocytes for measuring the production of tumour necrosis factor (TNF ), interleukin 6 (IL6) and IL12, while the anti-CD3 (1 µg/ml) and anti-CD28 (5 µg/ml) combination was used for T cell stimulation with the measuring of IL4 and interferon gamma (INF ) production. Twenty seven patients with RA and 23 healthy controls were studied. Results-The results showed a decreased IL6 (LPS stimulus 4-6 pg/ml) and IL12 (LPS stimulus 16-62 pg/ml) production in the RA patients. The maximal production of both cytokines was comparable with the normal controls. T cell stimulation showed a significant decreased INF production in the RA patients. Conclusions-These findings obtained in the WBC culture system are highly suggestive for a decreased TH-1 derived cytokine production by a diminished IL12 production in RA patients. Another possibility is that both IL12 and INF production in WBCs are inhibited by eventual circulating serum factors.

Adacolumn Selective Leukocyte Adsorption Apheresis in Patients with Active Ulcerative Colitis: Clinical Efficacy, Effects on Plasma IL-8, and Expression of Toll-like Receptor 2 on Granulocytes

“…We also looked at the effect of GMA on these cytokines in patients with active UC (n = 5) who received GMA. Blood samples were taken just before initiation of GMA and then from the Adacolumn inflow and outflow points 1 hr after the initiation of GMA and leukocytes were stimulated as above [27,28]. It appeared that when passed through the Adacolumn, leukocytes (unadsorbed fraction) have a reduced capacity to produce inflammatory cytokines (P < 0.05; Fig.…”

“…Cytokine production by peripheral blood leukocytes in whole blood culture cells was according to established methods [27,28]. In brief, whole blood cell cultures were done in 24 well-flat bottom culture plates (Nunc, Denmark).…”

A model of human whole blood lymphokine release for in vitro and ex vivo use

“…Three stimuli have been used, namely the lectin PHA, anti-CD3-antibodies and superantigens, In control experiments carried out in parallel in our study, PHA induced only a subset of lymphokines and clinical grade anti-CD3 (OKT-3, Orthoclone) did not induce any lymphokines at concentrations up to 2.5 ug/ml except when the blood was depleted of serum by washing (data not shown). This problem can apparently be overcome by eo-stimulation with anti-CD28 (Swaak et al, 1997;Lauw et al, 20(0).…”

“…The different types of whole blood lymphokine release assays have been shown to be of value for the evaluation of immune function in a broad range of patient groups, including cancer (Szopinski et al, 1999) , rheumatoid arthritis (Swaak et al, 1997), tuberculosis (van Crevel et al, 1999 ;Hussain et al, 2002), HIV (Hartung et al, 1998 ;Vingerhoets et al, 1998) and ankylosing spondylitis (Brand et al, 1997). However, since very different stimuli and experimental conditions were chosen, these studies are difficult to compare.…”

Change in cellular localization of a rheumatoid arthritis-related antigen (RA-A47) with downregulation upon stimulation by inflammatory cytokines in chondrocytes

“…Therefore, markers are expected to re¯ect both aspects of RA. As markers of in¯ammation, in¯ammatory cytokines such as interleukin (IL)-1 (Dayer and Burger, 1994), IL-6 (Maini et al, 1995;Swaak et al, 1997), and IL-8 (Maini et al, 1995), tumor necrosis factor (TNF) (Dayer and Burger, 1994), and interferon (IFN) (Ruschen et al, 1989); proteinases such as matrix metallo proteinase (MMP)-1 (Maeda et al, 1995), MMP-2 (Konttinen et al, 1999;Le et al, 1999), MMP-3 (Taylor et al, 1994), and MMP-9 (Yoshihara et al, 1995), plasminogen activator (Gotoh et al, 1997;Kamper et al, 1997;Kopeikina et al, 1997;Wallberg-Jonsson et al, 2000), and cathepsin-B and -H (Keyszer et al, 1998); and inhibitors such as tissue inhibitor of MMP (TIMP)-1 and TIMP-2 (Yoshihara et al, 1995), and plasminogen activator inhibitor Kopeikina et al, 1997;WallbergJonsson et al, 2000) are used. As markers of disruption of the joint, matrix components such as cartilage oligomeric matrix protein (COMP) (Wollheim, 1996;Marti et al, 1999) are considered for use.…”