Dynamic changes in cytokine levels in serum and synovial fluid following filtration leukocytapheresis therapy in patients with rheumatoid arthritis.

We attempted to determine whether various cytokine levels in the serum and synovial fluid (SF) of rheumatoid arthritis (RA) patients are influenced by the performance of filtration leukocytapheresis (LCP). The filtration LCP procedure that used a Cellsorba column (LCP group: n=22; responder subgroup: n=17, non-responder subgroup: n=5) or sham apheresis (control group; n=7) was repeated three times at 1-week intervals. Serum (LCP group, n=22; control group, n=7) and SF (LCP group, n=6; control group, n=3) samples were collected before and after LCP. Levels of tumor necrosis factor alpha (TNFalpha), interleukins (IL-1 beta, IL-2, IL-6, IL-8, IL-10, and IL-15), granulocyte-macrophage colony-stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), RANTES were measured by an enzyme-linked immunosorbent assay. Serum TNF alpha, IL-15, and RANTES were significantly reduced only in the LCP group. Serum IL-10 significantly increased only in the LCP group. In the LCP subgroup, serum IL-15, GM-CSF, and RANTES levels were reduced significantly, while serum IL-10 levels increased significantly only in the responder group after treatment. Serum TNF alpha levels were reduced significantly in both subgroups. Changes in serum IL-10 correlated positively with the improvement of patient's assessment of pain and global severity, and physician's assessment of global severity. These results indicate that the removal of leukocytes from the peripheral blood of RA patients provokes dynamic changes in some cytokine levels in the serum and/or synovial fluid. These changes may explain some of the mechanisms by which the articular symptoms are improved by filtration LCP.

Forearm heating band used in apheresis therapy that employs a positive-temperature coefficient polymer heater.

For standard apheresis therapy, blood is withdrawn from the ante-cubital vein of one arm and processed blood is returned to a vein of the opposite arm. For low-density lipoprotein apheresis or for the treatment of patients with Guillain-Barré syndrome, a sufficient quantity of blood is readily obtained by this method. In some patients with collagen diseases, however, it may be difficult to secure a reliable vein due to vasospasm or it may not be possible to obtain sufficient blood flow. We constructed a forearm heating band by employing a positive-temperature coefficient heater and evaluated the device to determine whether the application of heat to the forearm is effective in securing a sufficient quantity of blood in those patients with collagen disease (eight with systemic lupus erythematosus and one with multicentric Castleman's disease). Both forearms were heated by using this heating band, in addition to systemic warming with an electric blanket, starting 30 minutes before apheresis. The body surface temperature was sequentially monitored by employing a needle-type thermometer. The surface temperature of the heated area became constant at 37.6+/-0.3 degrees C within approximately 20 minutes (34.7+/-1.3 degrees C at the control site, P<0.001). It was found that this heating band makes it possible to obtain the quantity of blood that is necessary for apheresis and reduce the time required for the treatment. No adverse effects attributable to heating of the forearm were recognized.

Changes in CD4+ T lymphocyte subsets in circulating blood and synovial fluid following filtration leukocytapheresis therapy in patients with rheumatoid arthritis.

The purpose of this study is to determine the changes in CD4+ T lymphocyte subsets in the circulating blood and synovial fluid following filtration leukocytapheresis (LCP) therapy for patients with rheumatoid arthritis (RA). A Cellsorba column packed with polyester fibers was used for the removal of circulating leukocytes. For patients with RA, filtration LCP or sham procedures were performed 3 times with 1 week intervals between procedures. T lymphocyte surface markers in the peripheral blood and synovial fluid were measured by flow cytometry. The proportions of activated CD4+ T cells (CD4+DR+, CD4+CD25+, and CD4+CD71+) and CD4+CD29+ T cells increased significantly in the peripheral blood, but the counts of these cells were significantly reduced in the synovial fluid after 2 treatment sessions in the LCP group. No significant changes were observed in the proportion of these cells in the control group. Our findings suggest that filtration LCP may cause a redistribution of activated T cells from affected joints into the circulating blood.

Endothelin-1 enhances vascular cell adhesion molecule-1 expression in tumor necrosis factor alpha-stimulated vascular endothelial cells.

Vascular cell adhesion molecule-1 (VCAM-1) is a mononuclear leukocyte-selective adhesion molecule that is expressed in human vascular endothelial cells at sites of local inflammation. It participates in local endothelial-monocyte interactions during the initiation of atherosclerosis. In the present study, endothelin alone did not induce the surface expression and mRNA accumulation of VCAM-1 in human vascular endothelial cells, but inhibition of endogenous nitric oxide (NO) by N(G)-monomethyl-L-arginine enhanced the surface expression and mRNA accumulation of VCAM-1 stimulated by endothelin-1. It is conceivable that in human vascular endothelial cells, stimulation of an endothelin receptor results in the production of nitric oxide (NO), suppressing the expression of VCAM-1. Endothelin-1 enhanced the surface expression and mRNA accumulation of VCAM-1 in cells treated with tumor necrosis factor alpha (TNF-alpha). The enhancement by endothelin-1 may be explained by the inhibitory effect of TNF-alpha on endothelin-induced NO production. Pretreatment with BQ788 (an endothelin ET(B) receptor antagonist) or inhibitors of nuclear factor kappa B (NF-kappaB) activation completely diminished the synergistic enhancement of VCAM-1 expression by endothelin-1 in TNF-alpha-stimulated vascular endothelial cells, both at the protein and mRNA levels. These findings suggest that the synergistic enhancement of VCAM-1 expression by TNF-alpha and endothelin ET(B) receptor stimulation may be augmented by the induction of NF-kappaB binding activity in human vascular endothelial cells.

Filtration leukocytapheresis therapy in rheumatoid arthritis: a randomized, double-blind, placebo-controlled trial.

OBJECTIVE: To determine the efficacy and safety of filtration leukocytapheresis (LCP) for the treatment of rheumatoid arthritis (RA). METHODS: Twenty-five patients with drug-resistant RA were randomly assigned to undergo filtration LCP and 7 to undergo sham apheresis (control group) in a randomized, double-blind, placebo-controlled study. Three apheresis procedures were performed, with 1-week intervals between procedures. The efficacy of filtration LCP was evaluated according to the American College of Rheumatology definition of improvement in RA. Medications for each patient were unchanged for at least 6 months prior to enrollment and throughout the study. RESULTS: Tender joint counts, swollen joint counts, patient assessment of pain and global severity, physician assessment of global severity, and Health Assessment Questionnaire Disability Index were significantly improved in the LCP group compared with the control group (P < 0.05 for patient assessment of pain; P < 0.01 for all others). Seventy-nine percent of the patients in the LCP group exhibited significant overall improvement, while none of the patients in the control group were improved (P < 0.001). CONCLUSION: The results indicate that filtration LCP is an effective and well-tolerated treatment for patients with drug-resistant RA.

Evaluation of adsorption selectivity dextran sulfate bound cellulose beads for the removal of anti-DNA antibodies.

The purpose of this study was to clarify the basic adsorption selectivity characteristics of dextran sulfate (DS) columns (Selesorb, Kaneka Corporation, Osaka, Japan). Recovery rates of blood chemical components, hormones, coagulation factors, and antinuclear antibodies (anti-SS-A, SS-B, Sm, Scl-70, and RNP antibody) in vitro were assessed by mixing normal volunteers' or patients' sera with DS bound cellulose beads. For tested blood chemical components other than triglyceride and total cholesterol, the recovery rate was not changed significantly by incubation. No significant changes in hormone levels resulted from incubation. Among coagulation factors, the activities of antithrombin III, plasminogen, and factors V, VIII, IX, XI, and XII were significantly reduced by incubation. Among antinuclear antibodies tested, anti-SS-A and anti-RNP were absorbed to some extent, but not anti-SS-B, Sm, or Scl-70 antibodies. Taking into account these characteristics, apheresis therapy using a DS column should be performed.