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ABSTRACT: The distribution of folate receptor (FR)-β+ macrophages and their M1/M2 expression profiles were examined in osteoarthritis (OA) synovial tissues, and compared to those in rheumatoid arthritis (RA) synovial tissues and CD163+ macrophages in both OA and RA synovial tissues.
The phenotypes and fluorescein isothiocyanate (FITC)-folate uptake of FR-β+ synovial macrophages were analysed by flow cytometry. The distribution of FR-β+ macrophages in OA and RA synovial tissues was examined by immunofluorescent microscopy. Tumour necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), interleukin (IL)-10, and transforming growth factor (TGF)-β expression in FR-β+ macrophages was detected by double-immunostaining in both OA and RA synovial tissues.
FR-β+ macrophages were predominantly present in the synovial lining layer in OA patients. The proportion of CD163-FR-β+ cells in synovial mononuclear cells (MNCs) was increased in OA compared to RA synovial tissues. FR-β(high) macrophages from OA synovial tissues represented the majority of folic acid-binding cells. Although FR-β+ or CD163+ macrophages in the synovial tissues of OA and RA patients expressed a mixed pattern of M1 and M2 macrophage markers, there were more M2 markers expressing synovial macrophages in OA than in RA patients.
The distribution and M1/M2 expression profiles of FR-β+ synovial macrophages were different between OA and RA synovial tissues. Thus, the findings underscore that the M1/M2 paradigm using surface markers FR-β and CD163 is an oversimplification of macrophage subsets. Functional FR-β present on OA synovial macrophages provides a potential tool for the diagnosis and treatment of OA.
Scandinavian journal of rheumatology 01/2012; 41(2):132-40. · 2.51 Impact Factor
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ABSTRACT: To define the pathogenesis of pigmented villonodular synovitis (PVNS), by searching for highly expressed genes in primary synovial cells from patients with PVNS.
A combination of subtraction cloning and Southern colony hybridisation was used to detect highly expressed genes in PVNS in comparison with rheumatoid synovial cells. Northern hybridisation was performed to confirm the differential expression of the humanin gene in PVNS. Expression of the humanin peptide was analysed by western blotting and immunohistochemistry. Electron microscopic immunohistochemistry was performed to investigate the distribution of this peptide within the cell.
68 highly expressed genes were identified in PVNS. Humanin genes were strongly expressed in diffuse-type PVNS, but were barely detected in nodular-type PVNS, rheumatoid arthritis, or osteoarthritis. Humanin peptide was identified in synovium from diffuse-type PVNS, and most of the positive cells were distributed in the deep layer of the synovial tissue. Double staining with anti-humanin and anti-heat shock protein 60 showed that humanin was expressed mainly in mitochondria. Electron microscopy disclosed immunolocalisation of this peptide, predominantly around dense iron deposits within the siderosome.
Increased expression of the humanin peptide in mitochondria and siderosomes is characteristic of synovial cells from diffuse-type PVNS. Humanin is an anti-apoptotic peptide which is encoded in the mitochondrial genome. Present findings suggest that mitochondrial dysfunction may be the principal factor in pathogenesis of diffuse-type PVNS and that humanin peptide may play a part in the neoplastic process in this form of PVNS.
Annals of the Rheumatic Diseases 07/2005; 64(6):816-23. · 8.73 Impact Factor
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ABSTRACT: The aim of the present study was to evaluate levels of soluble CD 163 in sera and fluids from rheumatoid arthritis (RA) patients and elucidate the mechanism that regulates the shedding of CD163. Levels of soluble CD163 in sera and fluids from RA patients were examined by a sandwich enzyme immunoassay and Western blotting. To determine the effects of tissue inhibitors of metalloproteinase (TIMPs) on the shedding of CD163 from monocytes/macrophages, levels of soluble CD163 in cultures of monocytes/macrophages and the expression of CD163 on monocytes/macrophages in the presence or absence of TIMPs were examined by a sandwich enzyme immunoassay and flow cytometry, respectively. The clinical marker that was most associated with serum levels of soluble CD163 was levels of CRP. TIMP-3, but not TIMP-1 or TIMP-2, inhibited the shedding of CD163 from monocytes/macrophages. It was shown that serum levels of soluble CD163 are a sensitive and reliable marker to monitor activated macrophages in synovitis from RA patients and the results imply that the responsible proteinase for the shedding of CD163 is not a member of the matrix metalloproteinases, but is likely to be a member of ADAMs.
Clinical & Experimental Immunology 11/2002; 130(1):156-61. · 3.36 Impact Factor
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ABSTRACT: To investigate the expression of folate receptors (FR) and reduced folate carrier (RFC) and determine their relevance to methotrexate (MTX) transport in synovial mononuclear cells (SMC) from patients with rheumatoid arthritis (RA).
Levels of FR and RFC messenger RNA (mRNA) were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) in SMC from RA patients and peripheral blood mononuclear cells from healthy donors. Expression of FR-beta mRNA and protein was determined by Northern blot and Western blot analyses in RA SMC and monocyte/macrophage-lineage cells. FR-beta expression and folic acid binding capacity on the cell surface were examined by flow cytometric analysis and 3H-folic acid binding analysis. Studies of the inhibition of 3H-MTX uptake in the presence of unlabeled folic acid were performed to investigate the uptake of MTX through FR in RA SMC.
RT-PCR, Northern blot, and Western blot analyses showed that FR-beta mRNA and protein were expressed selectively in activated monocytes and CD14+ RA SMC. These cells exhibited folic acid binding capacity. Furthermore, the FR-beta protein was shown to have folic acid binding capacity. Uptake of 3H-MTX through RA SMC was significantly inhibited in the presence of unlabeled folic acid.
These results demonstrate that FR-beta expression is selectively elevated in RA synovial macrophages and suggest that MTX is transported through FR-beta in RA synovial macrophages. The findings suggest that folate antagonists with higher affinity for FR-beta would be useful in the treatment of RA.
Arthritis & Rheumatism 09/1999; 42(8):1609-16. · 7.87 Impact Factor
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ABSTRACT: It has been shown that cells with high affinity very late Ag (VLA)-integrins have up-regulated expression of a beta1-subunit epitope, which is detected by 15/7 mAb. In this study, we demonstrate that soluble VCAM-1 (sVCAM-1) exhibits chemotactic activity of T cells with high affinity VLA-4 against VCAM-1, such as Jurkat T cells and IL-2-dependent T cells. Moreover, we found that T cells in the synovial fluid show high basal migration in the absence of sVCAM-1, compared with peripheral blood T cells in patients with rheumatoid arthritis. Among T cells in the synovial fluid, CD45RO+ memory T cells, in response to sVCAM-1, showed a much higher than basal migratory response when compared with CD45RA+ naive cells, while no significant difference was observed between CD4+ and CD8+ T cells. The chemotactic activity of sVCAM-1 is inhibited in the presence of anti-VCAM-1 and anti-VLA-4, which interfered with the binding between VCAM-1 and VLA-4. Inhibition studies using various kinase inhibitors (C3 exoenzyme, KN62, and H7) show that Rho, Ca2+/calmodulin-dependent kinase II, and protein kinase C are involved in signal transduction in sVCAM-1-induced chemotaxis, respectively, whereas tyrosine kinase seems to play a lesser role, since genistein showed only partial inhibition of T cell chemotaxis. Western blot analysis using an anti-phospho-serine mAb (MO82) reveals that Ser82 in the vimentin is phosphorylated specifically by Ca2+/calmodulin-dependent kinase II through sVCAM-1 activation in the IL-2 dependent T cells. Collectively, by inducing migration and recruitment of T cells through several kinase activations, sVCAM-1 contributes to the development of the inflammation of synovial lesion.
The Journal of Immunology 12/1998; 161(9):4931-8. · 5.79 Impact Factor
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ABSTRACT: In this study, we made a mouse model for contact hypersensitivity (CH) using oxazolone as a contact allergen and examined the expression of interleukin-18 (IL-18) in the diseased skin sites at both mRNA and protein levels. In the kinetic study by semiquantitative RT-PCR, IL-18 mRNA was constitutively produced in normal murine skin but increased significantly at 12 h and peaked at 24 h in the ear skin of CH mice. A positive correlation was confirmed between the IL-18 mRNA signal and CH, as measured by mouse ear swelling response. Histologically, in situ hybridization showed that the IL-18 mRNA signal was weakly observed in the dermis but not the epidermis of normal skin, whereas the IL-18 mRNA signal was found intensively in the dermis, particularly in inflammatory cell areas. Using IL-18-specific antibody immunostaining, it was further found that IL-18 protein production had a histologic location similar to that of IL-18 mRNA in both normal and CH mice. The present study suggests that IL-18 may be implicated in the pathogenesis of murine CH.
Journal of Interferon & Cytokine Research 10/1998; 18(9):653-9. · 3.06 Impact Factor
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ABSTRACT: In order to establish what contributes to elevated levels of soluble CD14 (sCD14) in rheumatoid arthritis (RA) plasma, levels of sCD14 were compared in RA-paired plasma and synovial fluids and, further, in the culture supernatants of monocyte-rich fractions from patients with RA and healthy donors, and macrophage-rich fractions from RA synovial tissues. The results showed elevated sCD14 in RA synovial fluid in 9 of 16 paired samples and in RA macrophage-rich fractions, suggesting that elevated sCD14 in RA plasma might be due to the sCD14 production by RA synovial macrophages. From the molecular analysis of elevated sCD14, the proteolytic cleavage of membranous CD14 (mCD14) was important in accelerated sCD14 production. Lipopolysaccharides (LPS) at low concentrations and sCD14 increased the ICAM-1 expression on RA synovial fibroblasts. This result implies that in vivo RA synovial fibroblasts may be sensitive to LPS in the presence of sCD14 and LPS-binding protein (LBP).
Rheumatology International 02/1998; 17(6):237-43. · 1.88 Impact Factor
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ABSTRACT: It has been proven that interleukin-12 (IL-12) can modify Th1 and Th2 cell-mediated immune diseases by altering the development and cytokine production of the cells. In this study, we investigated the in vivo immunomodulatory effect of recombinant murine IL-12 on contact hypersensitivity, a Th1 cell-mediated disease. For this purpose, Balb/C mice were sensitized with 3% 4-ethyoxymethylene-2-phenyl-oxazol-5-one (OXAZ), and recombinant mouse IL-12 was given simultaneously during the induction phase. Contact allergy was then elicited by ear challenge with 1% OXAZ. We examined the mouse ear swelling response, in vivo cytokine gene expression in the skin and local lymph nodes, and in vitro cytokine production by the spleen lymphocytes. It was found that in vivo IL-12 treatment during the induction phase significantly enhanced the ear swelling response to OXAZ in sensitized mice. Moreover, remarkable mononuclear cell infiltration and edema and higher expression of Th1 cytokine mRNAs (IL-2 and interferon-gamma) in the skin lesion and local lymph nodes were observed in contact allergic mice with IL-12 treatment compared with contact allergic mice without IL-12 treatment. The expression of Th2 cytokine mRNA (IL-4) in the skin lesion and local lymph nodes, however, was largely downregulated, with no change in IL-5 mRNA in IL-12-treated contact allergic mice. We found, unexpectedly, that, similar to the effects on phytohemagglutinin (PHA) stimulated in vitro IL-2 and IFN-gamma production, PHA-induced in vitro IL-4 production was enhanced in the spleen lymphocytes from IL-12-treated contact allergic mice. Our results indicate that exogenous IL-12 enhanced contact hypersensitivity probably because of the in vivo promoting and suppressing effects of IL-12 on Th1 and Th2 gene expression, respectively.
Journal of Interferon & Cytokine Research 02/1998; 18(1):23-31. · 3.06 Impact Factor
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ABSTRACT: The present review focuses on the possible role of VCAM-1 expression on synovial fibroblast-like cells in the synovial lesion of rheumatoid arthritis (RA). The VCAM-1 expressing cells were mainly present in the synovial lining layer. The VCAM-1 expressing fibroblast-like cells also showed activity of uridine diphosphoglucose dehydrogenase, indicating that they are activated fibroblasts. VCAM-1 expressing T cells were also found in RA synovial fluids, where T lymphocytes show upregulation of alpha 4 beta 1 expression, and these T lymphocytes are able to bind to VCAM-1 in solid phase. Further experiments excluded the production of VCAM-1 protein in synovial fluid T lymphocytes and supported the idea that the soluble VCAM-1 was bound to the surface of synovial fluid T lymphocytes. We next planned to examine the effect of soluble VCAM-1 on T cell functions, by using recombinant soluble VCAM-1. The recombinant soluble VCAM-1 rendered synovial or peripheral T cells anergic to various stimuli. These findings imply that recombinant soluble VCAM-1 might be useful as a therapeutic tool to prevent abnormal immune response, since it binds to activated T lymphocytes with upregulation of alpha 4 beta 1, but not to resting T lymphocytes, and soluble VCAM-1 bound T lymphocytes become anergic.
Human Cell 10/1996; 9(3):187-92. · 1.27 Impact Factor
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ABSTRACT: Elevated levels of soluble vascular cell adhesion molecule-1 (sVCAM-1)/CD106 have been reported in synovial fluid (SF) from patients with rheumatoid arthritis (RA). In the present study, VCAM-1-positive lymphocytes were found in SF from RA patients. The data strongly suggest that sVCAM-1 might be bound to lymphocytes in SF. rsVCAM-1 in the fluid phase can bind to both SF lymphocytes and IL-2-dependent T cell lines with up-regulated expression and binding activity of VLA-4. Furthermore, proliferative responses of SF mononuclear cells (SFMC) with PHA, immobilized anti-CD3, or anti-CD2 and PMA were inhibited to various extents in the presence of rsVCAM-1, but only PMA-induced proliferative response of PBMC from normal individuals was inhibited notably in the presence of rsVCAM-1. rsVCAM-1 also drastically reduced IL-2 production of Jurkat leukemic T cells possessing high affinity VLA-4 with the stimulation of anti-CD3 and PMA, suggesting that the T cell hyporesponsiveness induced by rsVCAM-1 might stem from impairment of IL-2 production. These results indicate that sVCAM-1 provides a negative signal to T cell activation, probably by affecting the pathway of protein kinase C activation. Thus, binding of sVCAM-1 to SF lymphocytes might partly explain the anergic state of these lymphocytes.
The Journal of Immunology 04/1996; 156(6):2300-8. · 5.79 Impact Factor
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Bulletin of Environmental Contamination and Toxicology 01/1996; 55(6):789-95. · 1.02 Impact Factor
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ABSTRACT: To determine the effects of signalling through the DR molecule on synoviocytes from RA patients, the synovial adherent cells were incubated with anti-DR antibodies. After 24 h incubation, we found the formation of multinucleated giant cells in that culture. These multinucleated giant cells showed characteristics of monocyte-macrophage lineage cells and precursor of osteoclasts. Cyclohexamide inhibited the formation of multinucleated giant cells, but not the aggregation of synovial cells, suggesting that newly synthesized proteins are associated with the cell fusion. These results revealed a new mechanism in multinucleated giant cell formation.
Clinical & Experimental Immunology 12/1994; 98(2):257-63. · 3.36 Impact Factor
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ABSTRACT: To examine the effect of LY309887, an inhibitor of glycinamide ribonucleotide formyltransferase in de novo purine biosynthesis on murine type collagen-induced arthritis (CIA).
CIA was induced by immunization with bovine type II collagen in adjuvant. The expression of folate receptors was examined in dissected synovial tissues and bone marrow cells from arthritic and non-arthritic mice by the semi-quantitative reverse transcription-polymerase chain reaction. LY309887 was administered to CIA mice after the onset of arthritis. Mice were monitored for arthritis index for 21 days. Levels of IgG1 and IgG2a antibodies against bovine type II collagen were measured in sera from CIA mice with or without LY309887 treatment by the enzyme-linked immunosorbent assay. Histologic analyse were also performed in synovial tissues from arthritic joints with or without LY309887 treatment.
Levels of mRNA of folate receptor beta (FR-beta) were elevated in arthritic joints from CIA mice, compared with those in nonarthritic joints. The expression of mRNA of FR-beta was dominant in bone marrow cells of CIA mice. The administration of LY309887 suppressed the disease progression of CIA mice as defined by the lower arthritis index, and decreased production of serum IgG1 and IgG2a anti-type II collagen antibody, and the damage to cartilage or bone.
The administration of LY309887 was effective on CIA mice. It was suggested that LY309887 might be useful in the treatment of rheumatoid arthritis.
Clinical and experimental rheumatology 21(6):719-25. · 2.15 Impact Factor
