In order to estimate the dependence of CD4 and CD8 antigen expression upon the cell activation, t... more In order to estimate the dependence of CD4 and CD8 antigen expression upon the cell activation, their presence on resting and activated human T cells was investigated. Peripheral blood lymphocytes of healthy donors derived from late (TEl) and early (TEe) E rosettes were used as resting and presumably in vivo activated T cell subsets, respectively. Expression of the above markers on T cells stimulated in vitro with PHA was also examined. It was found that both lymphocyte subsets contained similar percentage of CD4+ cells, nevertheless, TEe subset was partially enriched in CD8+ cells. PHA stimulation induced in TEe and TEl subsets the considerable increase of proportion of phenotypical opposing cells, CD8+ or CD4+, respectively. The above changes coincided with the time of the maximal DNA synthesis (the 72nd h of cell stimulation).
* Cytokiny to glikoproteiny o niskim ciężarze cząsteczkowym, które są najczęściej wydzielane poza... more * Cytokiny to glikoproteiny o niskim ciężarze cząsteczkowym, które są najczęściej wydzielane poza komórkę i wpływają na jej metabolizm/czynność, wiążąc się do swoistych dla nich receptorów obecnych na komórkach (tego samego lub innego typu).
Expression of Fc gamma and Fc mu receptors on human peripheral blood T lymphocytes of two subsets... more Expression of Fc gamma and Fc mu receptors on human peripheral blood T lymphocytes of two subsets with high (E early rosette forming presumably in vivo activated cells, TEe) and low affinity of E receptors (E late rosette forming presumable resting cells, TEl) was investigated. Different distribution pattern of T gamma and T mu cells in the both examined T cell subsets was found. Thus TEe and TEl subsets have been partially enriched in T mu and T gamma cells, respectively. Furthermore, the results obtained in the PHA-stimulated system have shown that Fc mu receptors do not function as the markers of T cell activation. However, in opposition to this finding Fc gamma receptors may be the early activation markers but only of T cells originally bearing high-affinity E receptors.
High-affinity E-rosette receptor (EhR) is suggested to be an activation marker of human T lymphoc... more High-affinity E-rosette receptor (EhR) is suggested to be an activation marker of human T lymphocytes. The present study has been undertaken to estimate the dependence of EhR expression upon the T cell activation. To this aim the expression of EhR on T cells stimulated with PHA was examined. Nonsynchronized and arrested in G1 phase of cell cycle cultures of T lymphocytes were employed. The kinetics of expression of either de novo induced or reexpressed EhR was estimated by using two T cell subsets (originally EhR- and EhR+). In accordance with opinion of others, our results have shown that EhR is an activation marker of human T cells. Moreover, we have found that EhR is the marker of early and late activation stage because: its expression is induced in G1 phase of cell cycle, it continues to increase with the time of mitogen stimulation and the maximal level of EhR expression coincides with the time of the maximal RNA and DNA synthesis. Both induced de novo and reexpressed Eh receptors display the same expression kinetics.
Introduction: Oxidative modification of proteins affects their biological properties. Previously ... more Introduction: Oxidative modification of proteins affects their biological properties. Previously we have shown that hypochlorite (HOCl), the product of activated neutrophils, enhances protein immunogenecity. Collagen type II, a primary component of cartilage, is commonly used in the induction of arthritis in animals (CIA). The aim of this study was to examine whether HOCl may affect immunogenic, tolerogenic, and arthritogenic properties of collagen. Materials and methods: DBA/J mice were injected with either native (CNAT) or chlorinated collagen (CHOCl) to induce arthritis. The effect of chlorination on collagen properties was measured by evaluation of incidence and severity of CIA. Moreover, the concentration of serum anti-collagen IgG antibodies and myeloperoxidase (MPO) activity in inflamed joints was determined. Results: Mice immunized with CNAT in adjuvant developed arthritis (CIA) with an incidence of 69%. CNAT also exerted tolerogenic properties when injected intravenously either before or shortly after primary immunization, resulting in decreased incidence and severity of CIA, reduced MPO activity in inflamed joints, and lowered serum levels of anti-CNAT IgG anti-bodies. Chlorination of collagen significantly diminished its ability to induce CIA and to trigger generation of anti-CNAT IgG antibodies. Interestingly, chlorination did not affect tolerogenic properties of collagen administered prior to primary immunization with CNAT. Conclusions: These results suggest that chlorination of collagen may selectively affect functional epitopes of collagen. It is likely that in inflamed joints, neutrophil derived HOCl, in some circumstances, will destroy arthritogenic and immunogenic B cell epitopes, while regulatory T cell epitopes will be preserved.
DOAJ (DOAJ: Directory of Open Access Journals), Jun 1, 2011
Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reiche... more Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reicher w Warszawie Słowa kluczowe: cytokiny, degradacja chrząstki, resorpcja kości, nowe cele terapeutyczne.
Objectives: Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are chronic wasting, ... more Objectives: Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are chronic wasting, incurable rheumatic diseases of autoimmune background, in which T cells play a critical pathogenic role. Autologous adipose tissue-derived mesenchymal stem cells (ASCs) may represent an alternative therapeutic option for SLE and SSc patients, but the biology of these cells is poorly understood.
Background and objectives Adipose-derived mesenchymal stem cells (ASCs) have immunomodulatory pro... more Background and objectives Adipose-derived mesenchymal stem cells (ASCs) have immunomodulatory properties, but their activity is dependent on signals provided by local microenvironment. ASCs administration in rheumatoid arthritis (RA) did not bring consistent results, while in osteoarthritis (OA) it has beneficial effects. It is likely that proinflammatory milieu of rheumatoid joint affects ASCs activity. To test this hypothesis, the function of rheumatoid ASCs (RA-ASCs) and osteoarthritic ASCs (OA-ASCs) isolated from infrapatellar fat pad of the knee joint have been analysed. Materials and methods Infrapatellar fat pads were obtained from 29 RA and 12 OA patients undergoing total knee joint replacement surgery. Then, ASCs were isolated accordingly to routinely applied procedure. ASCs were treated or not with IFNγ or TNF. The expression of indoleamine 2,3-dioxygenase and heme oxygenase 1 mRNA was measured using quantitative PCR, while ASCs secretory potential was assessed by specific ELISAs. Rheumatoid synovial fibroblasts (RA-FLS) or peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured in ASC-conditioned media or co-cultured with ASCs. Next, cytokines secretion and cells proliferation were measured using ELISAs and bromodeoxyuridine/3H-thymidine proliferation assays, respectively. Results RA- and OA-ASCs activity in vitro is comparable, nonetheless some differences in spontaneous secretory activity and heme oxygenase 1 mRNA expression have been observed. Unstimulated RA- and OA-ASCs slightly inhibited PBMCs proliferation and induced IL-10 secretion but increased IL-17A production and failed to suppress release of other proinflammatory factors (TNF, IFNγ, RANTES) by PBMCs. Under TNF stimulation, RA- and OA-ASCs up-regulated IL-6 and MMP-3 secretion in RA-FLS cultures and IL-17A release by PBMCs. ASCs treatment with IFNγ or TNF did not intensify their anti-inflammatory activity. Conclusions Our study demonstrates that immunosuppressive function of RA- and OA-ASCs derived from infrapatellar fat pad is impaired. Possibly, it is due to the localisation of ASCs in the site of inflammation. Moreover, TNF triggers proinflammatory properties of ASCs which indicates that when administered to the rheumatoid joint, ASCs may shift from an anti-inflammatory to proinflammatory phenotype. Acknowledgements This work was supported by the Polish National Science Centre (grant no. 2011/01/N/NZ5/00932) and by the European Union funds (European Social Fund, Human Capital Programme).
DOAJ (DOAJ: Directory of Open Access Journals), Apr 1, 2011
Patogeneza reumatoidalnego zapalenia stawów. Część II-odpowiedź wrodzona, nowe cele terapeutyczne... more Patogeneza reumatoidalnego zapalenia stawów. Część II-odpowiedź wrodzona, nowe cele terapeutyczne Pathogenesis of rheumatoid arthritis. Part II-innate immunity, new therapeutic targets E Ew wa a K Ko on nt tn ny y Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reicher w Warszawie S Sł ło ow wa a k kl lu uc cz zo ow we e: : synowiocyty, komórki odporności wrodzonej, cząsteczki aktywujące, nowe cele terapeutyczne. K Ke ey y w wo or rd ds s: : synoviocytes, innate immunity cells, activating molecules, new therapeutic targets.
DOAJ (DOAJ: Directory of Open Access Journals), Apr 1, 2011
Leptyna i adiponektyna to klasyczne adipokiny produkowane przezbiałą tkankę tłuszczową. Mają one ... more Leptyna i adiponektyna to klasyczne adipokiny produkowane przezbiałą tkankę tłuszczową. Mają one działanie plejotropowe; ich rolębada się w takich chorobach, jak reumatoidalne zapalenie stawówczy choroba zwyrodnieniowa stawów. Dotąd nie wiadomo, jakidokładnie wpływ wywierają one na procesy chondrogenezyi osteoblastogenezy. Te dwa procesy są bardzo istotne z punktuwidzenia reumatoidalnego zapalenia stawów, gdyż w chorobie tejdochodzi do destrukcji chrząstki i kości stawowej. Istotne jestokreślenie, jaką rolę odgrywają adipokiny w reumatoidalnymzapaleniu stawów oraz w jaki sposób wpływają na różnicowaniekomórek mezenchymalnych. W niniejszej pracy przedstawionoobecną wiedzę na temat roli adiponektyny i leptyny w procesachosteogenezy i chondrogenezy (tab. I i II).
Background Most human peripheral blood monocytes strongly express surface CD14, and do not expres... more Background Most human peripheral blood monocytes strongly express surface CD14, and do not express CD16 (CD14 + +/ CD16-). A smaller group of monocytes express lower levels of CD14 and also express CD16 (CD14 + /CD16 +). This subgroup has different phagocytosing and antigen-presenting capabilities, and is expanded in a number of disease states. 1 Monocytederived macrophages and dendritic cells have important roles in the pathogenesis of both rheumatoid arthritis and SLE. Objectives To determine the percentage of circulating CD14 + / CD16 + monocytes in rheumatoid arthritis and systemic lupus erythematosus (SLE) and relate this to measures of disease activity and treatment. Methods Peripheral blood was sampled from 31 SLE patients, 19 rheumatoid arthritis patients and 19 healthy controls. The percentage of CD14 + /CD16 + monocytes was determined by immunofluorescence labelling and dual colour flow cytometry. Results The percentage of CD14 + /CD16 + monocytes was significantly reduced in rheumatoid arthritis (median = 4.90%) compared to normal subjects (median = 7.30%, p = 0.014), and in rheumatoid arthritis compared to SLE patients (median = 9.40%, p = 0.009). There was a trend towards a higher percentage of CD14 + /CD16 + monocytes in SLE compared to normal subjects, but this did not reach statistical significance. A negative correlation of percentage CD14 + /CD16 + monocytes with serum C-reactive protein level was observed (r =-0.461, p = 0.001). Drug treatment (including corticosteroid dose) did not correlate with the percentage of CD14 + /CD16 + monocytes in rheumatoid arthritis or SLE. Conclusion The percentage of CD14 + /CD16 + monocytes in rheumatoid arthritis is lower than in normal subjects and in SLE, and this may contribute to the pathogenesis of rheumatoid arthritis.
Wprowadzenie. U chorych na toczeń rumieniowaty układowy (ang. systemic lupus erythematosus-SLE) c... more Wprowadzenie. U chorych na toczeń rumieniowaty układowy (ang. systemic lupus erythematosus-SLE) często stwierdza się niedobór witaminy D. Spośród możliwych przyczyn tego stanu rozpatruje się: objęcie procesem chorobowym nerek, terapię z zastosowaniem kortykosteroidów lub leków przeciwmalarycznych, a także fotoprotekcję. Dotychczas nie w pełni poznany jest wpływ autoprzeciwciał skierowanych przeciwko kalcytriolowi na stężenie kalcydiolu w surowicy. Cel pracy. Ocena częstości występowania autoprzeciwciał skierowanych przeciwko kalcytriolowi w grupie chorych na SLE. Materiał i metodyka. Badaniem objęto grupę 37 chorych na SLE. Grupę kontrolną stanowiło 30 zdrowych dawców krwi. Przeciwciała oznaczano metodą immunoenzymatyczną. Wyniki. Autoprzeciwciała skierowane przeciwko kalcytriolowi wykryto u 3 pacjentów (8,1%) z SLE. Nie stwierdzono istotnej statystycznie różnicy w stężeniu kalcydiolu [25(OH)D 3 ] między chorymi, u których obecne były przeciwciała skierowane przeciwko kalcytriolowi, a osobami bez tych przeciwciał. Wnioski. W surowicy chorych na SLE obserwuje się autoprzeciwciała przeciwko kalcytriolowi, jednak ich obecność wydaje się nie wpływać na występowanie niedoboru witaminy D.
Background Classical adipokines, leptin and adiponectin, are thought to originate principally fro... more Background Classical adipokines, leptin and adiponectin, are thought to originate principally from white adipose tissue. Although both of them have been suggested to participate in pathogenesis of rheumatoid arthritis (RA), their role is still controversial [1]. Data concerning leptin contribution are inconsistent. Adiponectin was reported to exert mostly proinflammatory and prodestructive effects, but little is known about biological activities of its low (LMW) and high molecular (HMW) complexes. We have recently reported that not only rheumatoid synovial membrane (SM), but also articular adipose tissue (AAT) releases biologically active factors, including leptin and adiponectin [2]. However, it is not known whether periarticular subcutaneous adipose tissue (ScAT) shows similar secretory activity. Objectives (1) To compare the production of leptin and adiponectin by rheumatoid ScAT, AAT, and SM explants. (2) To investigate the effects of leptin and adiponectin on rheumatoid fibroblast-like synoviocytes (FLS) function. Methods Tissue specimens were obtained from the knee joints of 60 patients with established rheumatoid arthritis (RA) who were undergoing total joint replacement surgery. Tissue explants (100 mg/ml/well) were cultured in medium (DMEM) alone or treated for 18 h with recombinant human cytokines relevant to RA pathogenesis: IL-1β, TNF, interferon γ or IL-15, applied at 1, 10 or 40 ng/ml concentrations, respectively. After the treatment leptin and adiponectin concentrations were measured in tissue explants culture supernatants by ELISA. Moreover, RA FLS isolated from SM specimens were stimulated for 18 h with recombinant human leptin, HMW or LMW adiponectin (0.1-10 ng/ml), then the culture supernatants were collected and concentrations of proinflammatory cytokines (IL-6, IL-8) and connective tissue degrading enzyme, matrix metalloproteinase-3 (MMP-3), were measured using specific ELISA. Results Spontaneous leptin secretion by AAT, ScAT and SM explants was similar ($≈ $200 pg/100 mg), while SM secreted twice as much adiponectin as AAT and ScAT (mean ± SEM equal 7966±1016, 4649±340 and 4962±585 pg/100 mg, respectively). The release of both adipokines from SM did not change upon stimulation, while all applied stimuli raised their secretion from AAT and ScAT. In these conditions AAT and ScAT produced twice as much leptin but still less adiponectin than SM. Exogenously added adipokines exerted little (leptin), moderate (HMW adiponectin) or strong (LMW adiponectin) stimulatory effects on IL-6, IL-8 and MMP-3 secretion by RA FLS. Interestingly, the effect of LMW adiponectin (10 ng/ml) was comparable to that exerted by TNF. Conclusions Joint-associated adipose tissues (AAT and ScAT) are highly reactive to proinflammatory cytokines and upon stimulation their contribution to local adipokine pool is substantial. In RA joints adiponectin, especially of LMW, may contribute to synovitis and tissue destruction owing to potent up-regulation of IL-6, IL-8 and MMP-3 secretion by FLS. Acknowledgements This work was sponsored by grant No N N402 369938 from the Polish Ministry of Science and Higher Education. References Neumann E et al., Arthritis Rheum., 2011, 63:1159-69. Kontny E et al., Ann Rheum Dis., 2012, 71:262-267. Disclosure of Interest None Declared
A study was carried out to compare the phenotype, chondrogenic and osteogenic potential of adipos... more A study was carried out to compare the phenotype, chondrogenic and osteogenic potential of adipose mesenchymal stem cells (ASCs) derived from articular adipose tissue (AAT) and subcutaneous adipose tissue (ScAT) of rheumatoid arthritis (RA) patients. We also aimed to evaluate the role of leptin (LEP), low molecular weight adiponectin isoform (LMW), high molecular weight adiponectin isoform (HMW) and tumor necrosis factor (TNF) in ASCs differentiation. AAT and ScAT were obtained from RA patients undergoing total knee joint replacement surgery. ASCs were isolated and expanded in vitro. Cells phenotype was assesed by flow cytometry; leptin and adiponectin receptors mRNA expression by RT-PCR. Differentiation was performed in chondrogenic and osteogenic medium with or without LEP, LMW, HMW and TNF. After differentiation, expression of mRNA for Sox9, aggrecan (Agr), collagen 2a (Col2a) for chondrogenesis, and BMP-2, Runx-2 and osteopontin (OPN) for osteogenesis was evaluated by RT-PCR. Dickkopf-1 (DKK-1) and osteoprotegerin (OPG) proteins concentrations were measured in supernatants from osteogenic cultures using ELISA. AAT and ScAT-ASCs have the same CD105+CD90+CD73+CD45-CD34+/-CD19-CD14-phenotype. AAT-ASCs seem to be more susceptible to chondrogenesis and osteogenesis in vitro. Adipocytokines do not exert strong effect on ASCs differentiation, however HMW slightly increased expression of chondrogenesis markers mRNA in AAT-ASCs. TNF diminished expression of all chondrogenesis markers and OPN, but enhanced expression of BMP-2 and Runx-2 mRNA.
ObjectivesWhole body cryotherapy (WBC) is widely used in inflammatory diseases of the joints, inc... more ObjectivesWhole body cryotherapy (WBC) is widely used in inflammatory diseases of the joints, including rheumatoid arthritis (RA), but the mechanism(s) of its action is not fully understood. The aim of the study was to compare the effects of WBC and conventional rehabilitation (CR) on the clinical and immune status of RA patients.Material and methodsRheumatoid arthritis patients were classified into 2 groups according to the rehabilitation method used: the study group (CT, n = 25) and control group (CR, n = 25). To measure disease activity, the disease activity score (DAS28) was used, while to assess the morning stiffness and pain intensity, the visual analogue scale (VAS) was applied. Selected laboratory parameters, such as erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels, were also determined. The serum concentrations of pro- (interleukin 6 [IL-6], tumor necrosis factor a [TNF-a], macrophage migration inhibitory factor [MIF]) and anti-inflammatory (IL-10) c...
Background: Our previous data have proved that taurine chloramine (Tau-Cl), an endogenous molecul... more Background: Our previous data have proved that taurine chloramine (Tau-Cl), an endogenous molecule formed in activated neutrophils, normalizes pathological functions of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS)[1-3]. Moreover, we have recently reported that rheumatoid articular adipose tissue (AAT) produces factors activating FLS and thus may contribute to local pathological processes [4]. It is likely that Tau-Cl generated by infiltrating neutrophils may also affect cytokine secretion by joint-associated adipose tissue but this suggestion has not been verified yet. Objectives: To evaluate Tau-Cl effects on the production of select cytokines by rheumatoid synovial membrane (SM), AAT and subcutaneous periarticular adipose tissue (ScAT) explants. Methods: Tissue specimens were obtained from the knee joints of 64 patients with established RA who were undergoing total joint replacement surgery. Tissue explants (100 mg/ml/well) were treated for 18 h with 1 μg/ml of lipopolysaccharide (LPS) from Escherichia coli 055:B5 in the presence or absence of Tau-Cl (500 μM). After the treatment concentrations of select pro-(IL-1β, TNF, IL-6, IL-8) and anti-inflammatory (IL-10 and interleukin 1 receptor antagonist-IL-1Ra) cytokines were measured by specific ELISA. The Wilcoxon test was applied to evaluate the effects of LPS and Tau-Cl, while comparison between AAT, ScAT and SM cultures was done using Mann-Whitney U test. Results: Both AAT and ScAT produced spontaneously smaller amounts of all tested cytokines than SM. LPS vigorously raised release of cytokines from both adipose tissues with the fold increase within the range 10-20 for IL-1β, IL-1Ra, IL-6, and IL-8 or 50-100 for TNF and IL-10, while its stimulatory effect on SM was less pronounced (the fold increase within the range 2-20 and 30-40, respectively). Despite this, LPS-treated SM explants still released more cytokines than AAT and ScAT. In LPS-treated adipose tissues cultures Tau-Cl potently inhibited production of IL-1β, IL-6 and IL-1Ra but exerted weaker inhibitory effects on TNF and IL-8 release (% of inhibition within the range 60-70 and 25-30, respectively). In LPS-treated SM cultures Tau-Cl had no effect on TNF and IL-1Ra secretion but down-regulated IL-1β by 50%, while IL-6 and IL-8 by 25-30%. Importantly, Tau-Cl did not affect LPS-triggered IL-10 secretion by any tested tissues. Conclusions: We report for the first time that Tau-Cl is a potent inhibitor of pro-inflammatory cytokine secretion by joint-associated adipose tissues. Although this compound exerts weaker inhibitory effect on the release of proinflammatory cytokines by SM, its net local effect could be anti-inflammatory due to sparing IL-10 production and inhibition of mostly IL-1β (SM) or both IL-1β and IL-1Ra (AAT and ScAT). Thus, our results expand the spectrum of known anti-inflammatory activities of Tau-Cl and give further support to consider this compound as a promising candidate for RA treatment.
Conclusion The development of CIA and autoimmune responses in MRL/lpr mice were markedly suppress... more Conclusion The development of CIA and autoimmune responses in MRL/lpr mice were markedly suppressed by SX-5452 via inhibition of anti-CII/anti-DNA antibodies formation. Since SX-5452 exerts no overt toxicity after repeated administration in mice, it is an interesting candidate for therapeutic use in rheumatoid arthritis. THU0089 INHIBITION OF RHEUMATOID ARTHRIRTIS
In order to estimate the dependence of CD4 and CD8 antigen expression upon the cell activation, t... more In order to estimate the dependence of CD4 and CD8 antigen expression upon the cell activation, their presence on resting and activated human T cells was investigated. Peripheral blood lymphocytes of healthy donors derived from late (TEl) and early (TEe) E rosettes were used as resting and presumably in vivo activated T cell subsets, respectively. Expression of the above markers on T cells stimulated in vitro with PHA was also examined. It was found that both lymphocyte subsets contained similar percentage of CD4+ cells, nevertheless, TEe subset was partially enriched in CD8+ cells. PHA stimulation induced in TEe and TEl subsets the considerable increase of proportion of phenotypical opposing cells, CD8+ or CD4+, respectively. The above changes coincided with the time of the maximal DNA synthesis (the 72nd h of cell stimulation).
* Cytokiny to glikoproteiny o niskim ciężarze cząsteczkowym, które są najczęściej wydzielane poza... more * Cytokiny to glikoproteiny o niskim ciężarze cząsteczkowym, które są najczęściej wydzielane poza komórkę i wpływają na jej metabolizm/czynność, wiążąc się do swoistych dla nich receptorów obecnych na komórkach (tego samego lub innego typu).
Expression of Fc gamma and Fc mu receptors on human peripheral blood T lymphocytes of two subsets... more Expression of Fc gamma and Fc mu receptors on human peripheral blood T lymphocytes of two subsets with high (E early rosette forming presumably in vivo activated cells, TEe) and low affinity of E receptors (E late rosette forming presumable resting cells, TEl) was investigated. Different distribution pattern of T gamma and T mu cells in the both examined T cell subsets was found. Thus TEe and TEl subsets have been partially enriched in T mu and T gamma cells, respectively. Furthermore, the results obtained in the PHA-stimulated system have shown that Fc mu receptors do not function as the markers of T cell activation. However, in opposition to this finding Fc gamma receptors may be the early activation markers but only of T cells originally bearing high-affinity E receptors.
High-affinity E-rosette receptor (EhR) is suggested to be an activation marker of human T lymphoc... more High-affinity E-rosette receptor (EhR) is suggested to be an activation marker of human T lymphocytes. The present study has been undertaken to estimate the dependence of EhR expression upon the T cell activation. To this aim the expression of EhR on T cells stimulated with PHA was examined. Nonsynchronized and arrested in G1 phase of cell cycle cultures of T lymphocytes were employed. The kinetics of expression of either de novo induced or reexpressed EhR was estimated by using two T cell subsets (originally EhR- and EhR+). In accordance with opinion of others, our results have shown that EhR is an activation marker of human T cells. Moreover, we have found that EhR is the marker of early and late activation stage because: its expression is induced in G1 phase of cell cycle, it continues to increase with the time of mitogen stimulation and the maximal level of EhR expression coincides with the time of the maximal RNA and DNA synthesis. Both induced de novo and reexpressed Eh receptors display the same expression kinetics.
Introduction: Oxidative modification of proteins affects their biological properties. Previously ... more Introduction: Oxidative modification of proteins affects their biological properties. Previously we have shown that hypochlorite (HOCl), the product of activated neutrophils, enhances protein immunogenecity. Collagen type II, a primary component of cartilage, is commonly used in the induction of arthritis in animals (CIA). The aim of this study was to examine whether HOCl may affect immunogenic, tolerogenic, and arthritogenic properties of collagen. Materials and methods: DBA/J mice were injected with either native (CNAT) or chlorinated collagen (CHOCl) to induce arthritis. The effect of chlorination on collagen properties was measured by evaluation of incidence and severity of CIA. Moreover, the concentration of serum anti-collagen IgG antibodies and myeloperoxidase (MPO) activity in inflamed joints was determined. Results: Mice immunized with CNAT in adjuvant developed arthritis (CIA) with an incidence of 69%. CNAT also exerted tolerogenic properties when injected intravenously either before or shortly after primary immunization, resulting in decreased incidence and severity of CIA, reduced MPO activity in inflamed joints, and lowered serum levels of anti-CNAT IgG anti-bodies. Chlorination of collagen significantly diminished its ability to induce CIA and to trigger generation of anti-CNAT IgG antibodies. Interestingly, chlorination did not affect tolerogenic properties of collagen administered prior to primary immunization with CNAT. Conclusions: These results suggest that chlorination of collagen may selectively affect functional epitopes of collagen. It is likely that in inflamed joints, neutrophil derived HOCl, in some circumstances, will destroy arthritogenic and immunogenic B cell epitopes, while regulatory T cell epitopes will be preserved.
DOAJ (DOAJ: Directory of Open Access Journals), Jun 1, 2011
Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reiche... more Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reicher w Warszawie Słowa kluczowe: cytokiny, degradacja chrząstki, resorpcja kości, nowe cele terapeutyczne.
Objectives: Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are chronic wasting, ... more Objectives: Systemic lupus erythematosus (SLE) and systemic sclerosis (SSc) are chronic wasting, incurable rheumatic diseases of autoimmune background, in which T cells play a critical pathogenic role. Autologous adipose tissue-derived mesenchymal stem cells (ASCs) may represent an alternative therapeutic option for SLE and SSc patients, but the biology of these cells is poorly understood.
Background and objectives Adipose-derived mesenchymal stem cells (ASCs) have immunomodulatory pro... more Background and objectives Adipose-derived mesenchymal stem cells (ASCs) have immunomodulatory properties, but their activity is dependent on signals provided by local microenvironment. ASCs administration in rheumatoid arthritis (RA) did not bring consistent results, while in osteoarthritis (OA) it has beneficial effects. It is likely that proinflammatory milieu of rheumatoid joint affects ASCs activity. To test this hypothesis, the function of rheumatoid ASCs (RA-ASCs) and osteoarthritic ASCs (OA-ASCs) isolated from infrapatellar fat pad of the knee joint have been analysed. Materials and methods Infrapatellar fat pads were obtained from 29 RA and 12 OA patients undergoing total knee joint replacement surgery. Then, ASCs were isolated accordingly to routinely applied procedure. ASCs were treated or not with IFNγ or TNF. The expression of indoleamine 2,3-dioxygenase and heme oxygenase 1 mRNA was measured using quantitative PCR, while ASCs secretory potential was assessed by specific ELISAs. Rheumatoid synovial fibroblasts (RA-FLS) or peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured in ASC-conditioned media or co-cultured with ASCs. Next, cytokines secretion and cells proliferation were measured using ELISAs and bromodeoxyuridine/3H-thymidine proliferation assays, respectively. Results RA- and OA-ASCs activity in vitro is comparable, nonetheless some differences in spontaneous secretory activity and heme oxygenase 1 mRNA expression have been observed. Unstimulated RA- and OA-ASCs slightly inhibited PBMCs proliferation and induced IL-10 secretion but increased IL-17A production and failed to suppress release of other proinflammatory factors (TNF, IFNγ, RANTES) by PBMCs. Under TNF stimulation, RA- and OA-ASCs up-regulated IL-6 and MMP-3 secretion in RA-FLS cultures and IL-17A release by PBMCs. ASCs treatment with IFNγ or TNF did not intensify their anti-inflammatory activity. Conclusions Our study demonstrates that immunosuppressive function of RA- and OA-ASCs derived from infrapatellar fat pad is impaired. Possibly, it is due to the localisation of ASCs in the site of inflammation. Moreover, TNF triggers proinflammatory properties of ASCs which indicates that when administered to the rheumatoid joint, ASCs may shift from an anti-inflammatory to proinflammatory phenotype. Acknowledgements This work was supported by the Polish National Science Centre (grant no. 2011/01/N/NZ5/00932) and by the European Union funds (European Social Fund, Human Capital Programme).
DOAJ (DOAJ: Directory of Open Access Journals), Apr 1, 2011
Patogeneza reumatoidalnego zapalenia stawów. Część II-odpowiedź wrodzona, nowe cele terapeutyczne... more Patogeneza reumatoidalnego zapalenia stawów. Część II-odpowiedź wrodzona, nowe cele terapeutyczne Pathogenesis of rheumatoid arthritis. Part II-innate immunity, new therapeutic targets E Ew wa a K Ko on nt tn ny y Zakład Patofizjologii i Immunologii Instytutu Reumatologii im. prof. dr hab. med. Eleonory Reicher w Warszawie S Sł ło ow wa a k kl lu uc cz zo ow we e: : synowiocyty, komórki odporności wrodzonej, cząsteczki aktywujące, nowe cele terapeutyczne. K Ke ey y w wo or rd ds s: : synoviocytes, innate immunity cells, activating molecules, new therapeutic targets.
DOAJ (DOAJ: Directory of Open Access Journals), Apr 1, 2011
Leptyna i adiponektyna to klasyczne adipokiny produkowane przezbiałą tkankę tłuszczową. Mają one ... more Leptyna i adiponektyna to klasyczne adipokiny produkowane przezbiałą tkankę tłuszczową. Mają one działanie plejotropowe; ich rolębada się w takich chorobach, jak reumatoidalne zapalenie stawówczy choroba zwyrodnieniowa stawów. Dotąd nie wiadomo, jakidokładnie wpływ wywierają one na procesy chondrogenezyi osteoblastogenezy. Te dwa procesy są bardzo istotne z punktuwidzenia reumatoidalnego zapalenia stawów, gdyż w chorobie tejdochodzi do destrukcji chrząstki i kości stawowej. Istotne jestokreślenie, jaką rolę odgrywają adipokiny w reumatoidalnymzapaleniu stawów oraz w jaki sposób wpływają na różnicowaniekomórek mezenchymalnych. W niniejszej pracy przedstawionoobecną wiedzę na temat roli adiponektyny i leptyny w procesachosteogenezy i chondrogenezy (tab. I i II).
Background Most human peripheral blood monocytes strongly express surface CD14, and do not expres... more Background Most human peripheral blood monocytes strongly express surface CD14, and do not express CD16 (CD14 + +/ CD16-). A smaller group of monocytes express lower levels of CD14 and also express CD16 (CD14 + /CD16 +). This subgroup has different phagocytosing and antigen-presenting capabilities, and is expanded in a number of disease states. 1 Monocytederived macrophages and dendritic cells have important roles in the pathogenesis of both rheumatoid arthritis and SLE. Objectives To determine the percentage of circulating CD14 + / CD16 + monocytes in rheumatoid arthritis and systemic lupus erythematosus (SLE) and relate this to measures of disease activity and treatment. Methods Peripheral blood was sampled from 31 SLE patients, 19 rheumatoid arthritis patients and 19 healthy controls. The percentage of CD14 + /CD16 + monocytes was determined by immunofluorescence labelling and dual colour flow cytometry. Results The percentage of CD14 + /CD16 + monocytes was significantly reduced in rheumatoid arthritis (median = 4.90%) compared to normal subjects (median = 7.30%, p = 0.014), and in rheumatoid arthritis compared to SLE patients (median = 9.40%, p = 0.009). There was a trend towards a higher percentage of CD14 + /CD16 + monocytes in SLE compared to normal subjects, but this did not reach statistical significance. A negative correlation of percentage CD14 + /CD16 + monocytes with serum C-reactive protein level was observed (r =-0.461, p = 0.001). Drug treatment (including corticosteroid dose) did not correlate with the percentage of CD14 + /CD16 + monocytes in rheumatoid arthritis or SLE. Conclusion The percentage of CD14 + /CD16 + monocytes in rheumatoid arthritis is lower than in normal subjects and in SLE, and this may contribute to the pathogenesis of rheumatoid arthritis.
Wprowadzenie. U chorych na toczeń rumieniowaty układowy (ang. systemic lupus erythematosus-SLE) c... more Wprowadzenie. U chorych na toczeń rumieniowaty układowy (ang. systemic lupus erythematosus-SLE) często stwierdza się niedobór witaminy D. Spośród możliwych przyczyn tego stanu rozpatruje się: objęcie procesem chorobowym nerek, terapię z zastosowaniem kortykosteroidów lub leków przeciwmalarycznych, a także fotoprotekcję. Dotychczas nie w pełni poznany jest wpływ autoprzeciwciał skierowanych przeciwko kalcytriolowi na stężenie kalcydiolu w surowicy. Cel pracy. Ocena częstości występowania autoprzeciwciał skierowanych przeciwko kalcytriolowi w grupie chorych na SLE. Materiał i metodyka. Badaniem objęto grupę 37 chorych na SLE. Grupę kontrolną stanowiło 30 zdrowych dawców krwi. Przeciwciała oznaczano metodą immunoenzymatyczną. Wyniki. Autoprzeciwciała skierowane przeciwko kalcytriolowi wykryto u 3 pacjentów (8,1%) z SLE. Nie stwierdzono istotnej statystycznie różnicy w stężeniu kalcydiolu [25(OH)D 3 ] między chorymi, u których obecne były przeciwciała skierowane przeciwko kalcytriolowi, a osobami bez tych przeciwciał. Wnioski. W surowicy chorych na SLE obserwuje się autoprzeciwciała przeciwko kalcytriolowi, jednak ich obecność wydaje się nie wpływać na występowanie niedoboru witaminy D.
Background Classical adipokines, leptin and adiponectin, are thought to originate principally fro... more Background Classical adipokines, leptin and adiponectin, are thought to originate principally from white adipose tissue. Although both of them have been suggested to participate in pathogenesis of rheumatoid arthritis (RA), their role is still controversial [1]. Data concerning leptin contribution are inconsistent. Adiponectin was reported to exert mostly proinflammatory and prodestructive effects, but little is known about biological activities of its low (LMW) and high molecular (HMW) complexes. We have recently reported that not only rheumatoid synovial membrane (SM), but also articular adipose tissue (AAT) releases biologically active factors, including leptin and adiponectin [2]. However, it is not known whether periarticular subcutaneous adipose tissue (ScAT) shows similar secretory activity. Objectives (1) To compare the production of leptin and adiponectin by rheumatoid ScAT, AAT, and SM explants. (2) To investigate the effects of leptin and adiponectin on rheumatoid fibroblast-like synoviocytes (FLS) function. Methods Tissue specimens were obtained from the knee joints of 60 patients with established rheumatoid arthritis (RA) who were undergoing total joint replacement surgery. Tissue explants (100 mg/ml/well) were cultured in medium (DMEM) alone or treated for 18 h with recombinant human cytokines relevant to RA pathogenesis: IL-1β, TNF, interferon γ or IL-15, applied at 1, 10 or 40 ng/ml concentrations, respectively. After the treatment leptin and adiponectin concentrations were measured in tissue explants culture supernatants by ELISA. Moreover, RA FLS isolated from SM specimens were stimulated for 18 h with recombinant human leptin, HMW or LMW adiponectin (0.1-10 ng/ml), then the culture supernatants were collected and concentrations of proinflammatory cytokines (IL-6, IL-8) and connective tissue degrading enzyme, matrix metalloproteinase-3 (MMP-3), were measured using specific ELISA. Results Spontaneous leptin secretion by AAT, ScAT and SM explants was similar ($≈ $200 pg/100 mg), while SM secreted twice as much adiponectin as AAT and ScAT (mean ± SEM equal 7966±1016, 4649±340 and 4962±585 pg/100 mg, respectively). The release of both adipokines from SM did not change upon stimulation, while all applied stimuli raised their secretion from AAT and ScAT. In these conditions AAT and ScAT produced twice as much leptin but still less adiponectin than SM. Exogenously added adipokines exerted little (leptin), moderate (HMW adiponectin) or strong (LMW adiponectin) stimulatory effects on IL-6, IL-8 and MMP-3 secretion by RA FLS. Interestingly, the effect of LMW adiponectin (10 ng/ml) was comparable to that exerted by TNF. Conclusions Joint-associated adipose tissues (AAT and ScAT) are highly reactive to proinflammatory cytokines and upon stimulation their contribution to local adipokine pool is substantial. In RA joints adiponectin, especially of LMW, may contribute to synovitis and tissue destruction owing to potent up-regulation of IL-6, IL-8 and MMP-3 secretion by FLS. Acknowledgements This work was sponsored by grant No N N402 369938 from the Polish Ministry of Science and Higher Education. References Neumann E et al., Arthritis Rheum., 2011, 63:1159-69. Kontny E et al., Ann Rheum Dis., 2012, 71:262-267. Disclosure of Interest None Declared
A study was carried out to compare the phenotype, chondrogenic and osteogenic potential of adipos... more A study was carried out to compare the phenotype, chondrogenic and osteogenic potential of adipose mesenchymal stem cells (ASCs) derived from articular adipose tissue (AAT) and subcutaneous adipose tissue (ScAT) of rheumatoid arthritis (RA) patients. We also aimed to evaluate the role of leptin (LEP), low molecular weight adiponectin isoform (LMW), high molecular weight adiponectin isoform (HMW) and tumor necrosis factor (TNF) in ASCs differentiation. AAT and ScAT were obtained from RA patients undergoing total knee joint replacement surgery. ASCs were isolated and expanded in vitro. Cells phenotype was assesed by flow cytometry; leptin and adiponectin receptors mRNA expression by RT-PCR. Differentiation was performed in chondrogenic and osteogenic medium with or without LEP, LMW, HMW and TNF. After differentiation, expression of mRNA for Sox9, aggrecan (Agr), collagen 2a (Col2a) for chondrogenesis, and BMP-2, Runx-2 and osteopontin (OPN) for osteogenesis was evaluated by RT-PCR. Dickkopf-1 (DKK-1) and osteoprotegerin (OPG) proteins concentrations were measured in supernatants from osteogenic cultures using ELISA. AAT and ScAT-ASCs have the same CD105+CD90+CD73+CD45-CD34+/-CD19-CD14-phenotype. AAT-ASCs seem to be more susceptible to chondrogenesis and osteogenesis in vitro. Adipocytokines do not exert strong effect on ASCs differentiation, however HMW slightly increased expression of chondrogenesis markers mRNA in AAT-ASCs. TNF diminished expression of all chondrogenesis markers and OPN, but enhanced expression of BMP-2 and Runx-2 mRNA.
ObjectivesWhole body cryotherapy (WBC) is widely used in inflammatory diseases of the joints, inc... more ObjectivesWhole body cryotherapy (WBC) is widely used in inflammatory diseases of the joints, including rheumatoid arthritis (RA), but the mechanism(s) of its action is not fully understood. The aim of the study was to compare the effects of WBC and conventional rehabilitation (CR) on the clinical and immune status of RA patients.Material and methodsRheumatoid arthritis patients were classified into 2 groups according to the rehabilitation method used: the study group (CT, n = 25) and control group (CR, n = 25). To measure disease activity, the disease activity score (DAS28) was used, while to assess the morning stiffness and pain intensity, the visual analogue scale (VAS) was applied. Selected laboratory parameters, such as erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels, were also determined. The serum concentrations of pro- (interleukin 6 [IL-6], tumor necrosis factor a [TNF-a], macrophage migration inhibitory factor [MIF]) and anti-inflammatory (IL-10) c...
Background: Our previous data have proved that taurine chloramine (Tau-Cl), an endogenous molecul... more Background: Our previous data have proved that taurine chloramine (Tau-Cl), an endogenous molecule formed in activated neutrophils, normalizes pathological functions of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS)[1-3]. Moreover, we have recently reported that rheumatoid articular adipose tissue (AAT) produces factors activating FLS and thus may contribute to local pathological processes [4]. It is likely that Tau-Cl generated by infiltrating neutrophils may also affect cytokine secretion by joint-associated adipose tissue but this suggestion has not been verified yet. Objectives: To evaluate Tau-Cl effects on the production of select cytokines by rheumatoid synovial membrane (SM), AAT and subcutaneous periarticular adipose tissue (ScAT) explants. Methods: Tissue specimens were obtained from the knee joints of 64 patients with established RA who were undergoing total joint replacement surgery. Tissue explants (100 mg/ml/well) were treated for 18 h with 1 μg/ml of lipopolysaccharide (LPS) from Escherichia coli 055:B5 in the presence or absence of Tau-Cl (500 μM). After the treatment concentrations of select pro-(IL-1β, TNF, IL-6, IL-8) and anti-inflammatory (IL-10 and interleukin 1 receptor antagonist-IL-1Ra) cytokines were measured by specific ELISA. The Wilcoxon test was applied to evaluate the effects of LPS and Tau-Cl, while comparison between AAT, ScAT and SM cultures was done using Mann-Whitney U test. Results: Both AAT and ScAT produced spontaneously smaller amounts of all tested cytokines than SM. LPS vigorously raised release of cytokines from both adipose tissues with the fold increase within the range 10-20 for IL-1β, IL-1Ra, IL-6, and IL-8 or 50-100 for TNF and IL-10, while its stimulatory effect on SM was less pronounced (the fold increase within the range 2-20 and 30-40, respectively). Despite this, LPS-treated SM explants still released more cytokines than AAT and ScAT. In LPS-treated adipose tissues cultures Tau-Cl potently inhibited production of IL-1β, IL-6 and IL-1Ra but exerted weaker inhibitory effects on TNF and IL-8 release (% of inhibition within the range 60-70 and 25-30, respectively). In LPS-treated SM cultures Tau-Cl had no effect on TNF and IL-1Ra secretion but down-regulated IL-1β by 50%, while IL-6 and IL-8 by 25-30%. Importantly, Tau-Cl did not affect LPS-triggered IL-10 secretion by any tested tissues. Conclusions: We report for the first time that Tau-Cl is a potent inhibitor of pro-inflammatory cytokine secretion by joint-associated adipose tissues. Although this compound exerts weaker inhibitory effect on the release of proinflammatory cytokines by SM, its net local effect could be anti-inflammatory due to sparing IL-10 production and inhibition of mostly IL-1β (SM) or both IL-1β and IL-1Ra (AAT and ScAT). Thus, our results expand the spectrum of known anti-inflammatory activities of Tau-Cl and give further support to consider this compound as a promising candidate for RA treatment.
Conclusion The development of CIA and autoimmune responses in MRL/lpr mice were markedly suppress... more Conclusion The development of CIA and autoimmune responses in MRL/lpr mice were markedly suppressed by SX-5452 via inhibition of anti-CII/anti-DNA antibodies formation. Since SX-5452 exerts no overt toxicity after repeated administration in mice, it is an interesting candidate for therapeutic use in rheumatoid arthritis. THU0089 INHIBITION OF RHEUMATOID ARTHRIRTIS
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