Takaaki Fukuda

Effects of α-Glucosylhesperidin, a Bioactive Food Material, on Collagen-Induced Arthritis in Mice and Rheumatoid Arthritis in Humans

Hesperidin (Hsp) is an abundant flavonoid in citrus fruits, and the oral administration of Hsp has been recently reported to suppress collagen-induced arthritis in mice. Therefore, we sought to determine whether α-glucosylhesperidin (Hsp-G), which is an Hsp derivative with enhanced water-solubility, is effective on treating arthritis in both mice and humans. Hsp-G was orally administered to mice with collagen-induced arthritis, and its effects were evaluated clinically and histologically. Oral administration of Hsp-G improved collagen-induced arthritis when administered before the onset of arthritis as well as when administered after its onset. A decrease in tumor necrosis factor-α production was found to cause this improvement. In the human study, 19 patients with rheumatoid arthritis (RA) were enrolled in a 12-week double-blind, placebo-controlled trial. Patients were administered beverages containing 3 g Hsp-G (n = 9) or placebo (n = 10) every morning for the duration of the 3-month trial. Additionally, patients received standard therapy from a physician every 4 weeks. As a result, 3 of 9 patients in the Hsp-G group improved, while only 1 of 10 patients in the placebo group improved; this was in accordance with the American College of Rheumatology criteria. The present study revealed that the food material Hsp-G was effective when administered with standard anti-rheumatoid therapy in ameliorating RA in mice and humans without any adverse effects and may improve the quality of life for patients with RA as a complementary/alternative medicine.

Soluble interleukin-18 receptor complex is a novel biomarker in rheumatoid arthritis

IntroductionThere has been no report in the literature of a soluble form of interleukin (IL)-18 receptor α (IL-18Rα). In this study, we evaluated the levels and characteristics of soluble IL-18Rα (sIL-18Rα) in the sera of patients with rheumatoid arthritis (RA) and compared these results to control populations.MethodsThe sIL-18Rα complex was isolated from pooled human blood serum using an anti-IL-18Rα monoclonal antibody affinity column. The purified sIL-18Rα was then examined using Western blot analysis and used in experiments to evaluate the effects on an IL-18-responsive natural killer (NK) human cell line, NK0. An enzyme-linked immunosorbent assay was developed, and sera from 145 patients with RA, 6 patients with adult-onset Stills disease, 31 patients with osteoarthritis (OA), 39 patients with systemic lupus erythematosus (SLE) and 67 controls were tested, along with levels of immunoglobulin M, rheumatoid factor, anticyclic citrullinated peptide antibody, IL-18, IL-13 and interferon (IFN)-γ. Area under the receiver operating characteristic curve (ROC-AUC) analysis was used to evaluate the diagnostic utility of the sIL-18Rα complex.ResultsThe isolated sIL-18Rα complex can be associated with IL-18 and the soluble form of the IL-18Rβ chain. The sIL-18Rα complex bound to the surface to the NK0 cell line, antagonized the stimulatory effects of IL-18 and IL-2 on the NK0 cell line and inhibited IFN-γ production by the cells. The serum levels of sIL-18Rα complex in RA (186.0 ± 33.5 ng/mL, n = 145) and adult-onset Stills disease (98.2 ± 8.9 ng/mL, n = 6) were significantly (P < 0.001) higher than those in the healthy controls (52.3 ± 8.5 ng/mL, n = 67), OA (38.6 ± 5.4 ng/mL, n = 31), SLE (44.6 ± 3.2 ng/mL, n = 39). The serum level of sIL-18Rα complex was not significantly different between RA and adult-onset Stills disease patients. The serum levels of IL-18, IL-13 and IFN-γ in the RA patients were significantly (P < 0.01) higher than in OA and SLE patients as well as healthy controls. ROC-AUC analysis of the serum concentration of sIL-18Rα indicated that it was significantly diagnostic of RA. Moreover, a tumor necrosis factor inhibitor, etanercept, significantly (P < 0.0001) decreased levels of sIL-18Rα in the sera of 29 RA patients 6 months after treatment.ConclusionsThe sIL-18Rα complex could be a potentially useful biomarker for the diagnosis of RA.

Expression of membrane-type 1 matrix metalloproteinase in rheumatoid synovial cells

Membrane‐type 1 matrix metalloproteinase (MT1‐MMP) is thought to be a putative regulator of pro‐gelatinase A (MMP‐2) in the rheumatoid synovium. In this study, we examined the effects of IL‐1β, one of the inflammatory cytokines, on the expression of MT1‐MMP and the activation of pro‐MMP‐2 using rheumatoid synovial cells. We also studied the effects of KE‐298 (2‐acetylthiomethyl‐4‐(4‐methylphenyl)‐4‐oxobutanoic acid), a new disease‐modifying anti‐rheumatic drug (DMARD), on MT1‐MMP expression of rheumatoid synovial cells. Type B synovial cells (fibroblast‐like synovial cells) were cultured with KE‐298 (25–100 µg/ml) in the presence of IL‐1β for 48 h. Activation of pro‐MMP‐2 secreted from synovial cells was analysed by gelatin zymography. Reverse transcription–polymerase chain reaction (RT–PCR) methods were used to detect MT1‐MMP mRNA. MT1‐MMP protein expression on synovial cells was examined by anti‐MT1‐MMP immunoblot. An active form of MMP‐2 was demonstrated in the culture media conditioned by IL‐1β‐stimulated synovial cells. In addition, MT1‐MMP mRNA and protein expression of rheumatoid synovial cells were increased by IL‐1β treatment. KE‐298 blocked this IL‐1β‐induced pro‐MMP‐2 activation and MT1‐MMP expression, but did not affect IL‐1β‐induced tissue inhibitor of metalloproteinase‐2 (TIMP‐2) secretion from rheumatoid synovial cells. These findings indicate that activation of rheumatoid synovial cells by IL‐1β results in the induction of MT1‐MMP expression. Given that MT1‐MMP promotes matrix degradation by activating pro‐MMP‐2, these results suggest a novel mechanism whereby cytokine may contribute to articular destruction in rheumatoid arthritis (RA). KE‐298 may prevent this process by down‐regulating MT1‐MMP expression.

HTLV-I associated arthritis: characteristics of an HTLV-I virus infected T cell line from synovial fluid.

A T cell line from mononuclear cells in the synovial fluid of a patient with polyarthritis was established. The T cell line reacted with serum samples positive for antibodies to human T cell lymphotropic virus type I (HTLV-I) and with monoclonal antibody to HTLV-I p19. In Southern blotting with an env-pX-LTR HTLV-I probe and digestion of T cell line DNA with the restriction enzymes ClaI, DraI, and PstI generated fragments that were identical to those found in two HTLV-I infected T cell lines established from adult T cell leukaemia or HTLV-I associated myelopathy. The T cell line expressed CD2, CD3, CD4, CD45RA, CD29, HLA-DR, CD25, and CD26 antigens, but not CD8 and CD20 antigens. Large amounts of interleukin 6, interferon gamma, and tumour necrosis factor alpha were secreted in the culture supernatants of this cell line. This line helped immunoglobulin production by B cells, but not K562, Raji, and synovial cell lysis.

Sustainable Efficacy of Switching From Intravenous to Subcutaneous Tocilizumab Monotherapy in Patients With Rheumatoid Arthritis

To evaluate the efficacy and safety of switching from intravenous (IV) tocilizumab (TCZ) to subcutaneous (SC) TCZ monotherapy in rheumatoid arthritis patients.

Humoral immune responses to CTL epitope peptides from tumor-associated antigens are widely detectable in humans: A new biomarker for overall survival of patients with malignant diseases

Both cellular and humoral immune responses are crucial to induce potent anti-tumor immunity, but most of currently conducted peptide-based cancer vaccines paid attention to cellular responses alone, and none of them are yet approved as a therapeutic modality against cancer patients. We investigated humoral immune responses to CTL epitope peptides derived from tumor-associated antigens in healthy donors and patients with various diseases to facilitate better understanding of their distribution patterns and potential roles. Bead-based multiplex assay, ELISA, and Western blotting were used to measure immunoglobulins reactive to each of 31 different CTL epitope peptides. Importantly, the sums of anti-peptide IgG levels specific to 31 CTL epitope peptides were well correlated with better overall survival (OS) in patients with malignant diseases. Our results suggested that humoral immune responses to CTL epitope peptides were widely detectable in humans. Measurement of immunoglobulins specific to CTL epitope peptides may provide a new biomarker for OS of patients with malignant diseases, although it still remains to be determined whether the correlations between humoral immune responses to epitope peptides and OS are observed only for the CTL epitopes used, or also for other panels of peptides. Quantity of circulating IgG reactive to these peptides was also discussed.

Successful treatment of macrophage activation syndrome in a patient with dermatomyositis by combination with immunosuppressive therapy and plasmapheresis

Macrophage activation syndrome (MAS), also known as secondary hemophagocytic lymphohistiocytosis, is mediated by cytokine overproduction from excessive activation of T lymphocytes and macrophages. We present a dermatomyositis patient with MAS, caused by hypercytokinemia. The combination of tacrolimus and plasma exchange therapy was effective in this case for treating MAS. This combination therapy is especially useful for MAS refractory to steroids.

Bronchiolitis obliterans organizing pneumonia in a patient with primary biliary cirrhosis and rheumatoid arthritis treated with prednisolone

A 62-year-old woman with primary biliary cirrhosis (PBC) and rheumatoid arthritis (RA) was examined at our hospital for a 2-week history of non-resolving fever, cough and malaise. Her chest radiograph revealed left lower lobe opacity. Various kinds of antibiotics were not effective and transbronchial biopsy revealed non-specific inflammatory alveolar lesions. Chest radiograph at 14 days after admission revealed migration of the pulmonary opacity, suggesting bronchiolitis obliterans organizing pneumonia (BOOP), which responded well clinically and radiologically to oral corticosteroid therapy. BOOP may be one of the possible non-hepatic complications of PBC especially in patients associated with other connective tissue diseases.

Expression of cyclooxygenase-1 (COX-1) in labial salivary glands of Sjögren's syndrome

COX plays an important role in inflammatory diseases such as rheumatoid arthritis. To determine the role of COX in Sjögrens syndrome (SS), we examined COX expression in the salivary glands of SS patients. We examined 15 patients with SS and two normal subjects. Labial salivary gland tissue samples were analysed immunohistochemically using anti‐COX‐1 and COX‐2 antibodies. All biopsy samples from 15 patients with SS were stained for COX‐1. In contrast, COX‐1 immunostaining was not detected in normal salivary gland tissues. Co‐expression of COX‐1 and CD68 was confirmed by mirror section technique and double antibody immunostaining. This finding indicated that COX‐1‐expressing cells in SS salivary glands were infiltrating macrophages. In contrast to COX‐1 staining, only a little COX‐2 immunostaining was observed in salivary gland tissues from SS patients. These data suggest that COX‐1 expression on infiltrating macrophages may contribute to the inflammatory process of salivary glands in SS.

Impact of methotrexate dose on efficacy of adalimumab in Japanese patients with rheumatoid arthritis: Results from registered data analyses

Abstract Objective: Upper limit of methotrexate (MTX) for patients with rheumatoid arthritis (RA) was recently increased from 8 to 16 mg/week in Japan. We therefore examined the effect of concomitant MTX dose on the efficacy of adalimumab (ADA) in clinical practice. Method: Sixty-one consecutive RA patients treated with ADA were followed for minimum 52 weeks and retrospectively compared by MTX dose; patients receiving concomitant MTX of 10 mg/week or more (MTX ≥10 mg group) and <10 mg/week (MTX <10 mg group). Disease activity and remission were evaluated by the disease activity score 28 (DAS28) criteria. Results: The MTX ≥10 mg group consistently showed better improvement in DAS28 and resulted in more patients (52.8%) with DAS28-remission compared with the MTX <10 mg group (26.1%). Multivariate analysis showed that MTX ≥10 mg had a significant effect on DAS28 remission with odds ratio of 5.12. ADA retention rate was 72.2% in MTX ≥10 mg group compared with 52.0% in MTX <10 mg group. Discontinuation of ADA due to adverse events were comparable in the MTX ≥10 mg and MTX <10 mg groups (11.1% vs. 12.0%). Conclusions: These findings support the critical role of concomitant MTX in the efficacy of ADA, and recommend use of MTX ≥10 mg in Japanese RA patients.