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OBJECTIVE: The signaling lymphocytic activation molecule family of receptors (SLAMF) is involved in the activation of T cells and plays important roles in the pathogenesis of autoimmune diseases. The purpose of this study is to observe the expression of SLAMF3 on CD4 + T cells and its effect on the differentiation of T helper 17 (Th17) in primary Sjögren's syndrome (pSS). Furthermore, we found iguratimod (IGU) could effectively reverse the aberrant Th17 differentiation through JAK1/STAT3 signaling. METHODS: Peripheral blood mononuclear cells from 40 pSS and 40 healthy control subjects were enrolled for analysis of expression of SLAMF3 on CD4 + T and Th17 cells by flow cytometry. Serum IL-17 and SLAMF3 were detected by ELISA assay. Labial biopsies from 20 pSS patients and 20 non-pSS controls were performed immunohistochemical for staining expression of CD4, IL-17, and SLAMF3. Under the priming conditions with anti-CD3/CD28 or CD3/SLAMF3 antibodies on CD4 + T cells extracted from pSS and controls, the proportion of Th17 cells in CD4 + T cells and the amount of soluble IL-17A were assessed by flow cytometry and ELISA. Furthermore, RNA sequencing was performed for the transcriptomics study. Additionally, RNA level of RORγt and IL-17A and the protein level of RORγt, p-JAK1 and p-STAT3, were detected by real-time PCR and western blot. RESULTS: The expression levels of SLAMF3 on CD4 + T and Th17 cells in the peripheral blood and salivary glands in pSS patients were significantly elevated than that in control groups. The serum IL-17A and SLAMF3 in pSS patients were much higher compared with the control group. Although co-stimulation of CD3/SLAMF3 could promote CD4 + T cells differentiate into Th17 cells both in pSS and controls, the CD4 + T cells from pSS have a more sensitive response in Th17 differentiation with the SLAMF3 stimulation. Transcriptomics results showed the CD3/SLAMF3 stimulation caused the activation of Th17 signaling and JAK1/STAT3 pathway. Quantitative PCR and western blotting confirmed the IGU (iguratimod), which is a safe clinical drug in treatment of autoimmune diseases, effectively reversed the increased Th17 proportion, the expression levels of RORγt, pJAK1, and pSTAT3 caused by CD3/SLAMF3 stimulation. CONCLUSION: SLAMF3 upregulates Th17 cell differentiation of CD4 + T cells and IL-17A secretion through enriching RORγt and activating the transcriptomics participating in the pathogenesis of primary Sjögren's syndrome. IGU could inhibit the process through this therapeutic target in pSS.
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Interleucina-17 , Síndrome de Sjögren , Humanos , Diferenciación Celular , Interleucina-17/metabolismo , Janus Quinasa 1/metabolismo , Leucocitos Mononucleares/metabolismo , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/genética , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo , Factor de Transcripción STAT3/metabolismo , Células Th17RESUMEN
OBJECTIVES: Neuropsychiatric systemic lupus erythematosus (NPSLE) is a serious phenotype of systemic lupus erythematosus (SLE). The disturbance of neuron-microglia crosstalk is recently revealed in many neuropsychiatric diseases but was not well studied in NPSLE. We found glucose regulatory protein 78 (GRP78), a marker of endoplasmic reticulum stress, was significantly increased in the cerebrospinal fluid (CSF) of our NPSLE cohort. We, therefore, investigated whether GRP78 can act as a mediator between the neuron-microglia crosstalk and is involved in the pathogenic process of NPSLE. METHODS: Serum and CSF parameters were analyzed in 22 NPSLE patients and controls. Anti-DWEYS IgG was injected intravenously into mice to establish a model of NPSLE. Behavioral assessment, histopathological staining, RNA-seq analyses, and biochemical assays were performed to examine the neuro-immunological alterations in the mice. Rapamycin was intraperitoneally administered to define the therapeutic effect. RESULTS: The level of GRP78 was elevated significantly in the CSF of the patients with NPSLE. An increase in GRP78 expression, accompanied by neuroinflammation and cognitive impairment, was also found in the brain tissues of the NPSLE model mice induced by anti-DWEYS IgG deposition on hippocampal neurons. In vitro experiments demonstrated that anti-DWEYS IgG could stimulate neurons to release GRP78, which activated microglia via TLR4/MyD88/NFκB pathway to produce more pro-inflammatory cytokines and promote migration and phagocytosis. Rapamycin ameliorated GRP78-inducing neuroinflammation and cognitive impairment in anti-DWEYS IgG-transferred mice. CONCLUSION: GRP78 acts as a pathogenic factor in neuropsychiatric disorders via interfering neuron-microglia crosstalk. Rapamycin may be a promising therapeutic candidate for NPSLE.
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Chaperón BiP del Retículo Endoplásmico , Lupus Eritematoso Sistémico , Vasculitis por Lupus del Sistema Nervioso Central , Animales , Ratones , Chaperón BiP del Retículo Endoplásmico/metabolismo , Glucosa , Inmunoglobulina G , Microglía , Enfermedades Neuroinflamatorias , Neuronas , HumanosRESUMEN
Chemokine-like factor (CKLF)-like MARVEL transmembrane domain containing family member 6 (CMTM6), which is a key regulator of programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) signaling in patients with primary Sjögren's syndrome (pSS). In this study, we analyzed the serum levels of CMTM6, PD-1, and PD-L1 in 50 patients with pSS, 42 patients with non-pSS (simply dry mouth and/or eyes symptoms) and 50 healthy controls (HC). The expression of CMTM6, PD-1, and PD-L1 in labial glands of the same 50 pSS patients and 42 non-pSS patients were assessed by immunohistochemistry (IHC). The clinical significance of CMTM6, PD-1, and PD-L1 were analyzed. We found that levels of CMTM6, PD-L1 as well as PD-1 in sera were all increased significantly in patients with pSS compared with non-pSS controls and HC. Serum CMTM6 level showed significantly correlation with PD-L1, PD-1, as well as clinical laboratory indicators and disease activity of pSS patients. CMTM6, PD-1, and PD-L1 expression in labial glands was also higher significantly in pSS patients than non-pSS controls. pSS patients with higher CM grade or ESSDAI score have higher CMTM6, PD-L1, and PD-1 expression in labial glands. These results suggest that CMTM6 may affect peripheral tolerance and lymphocytes activation by PD-1/PD-L1 pathway in sera and target tissue in pSS.
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Síndrome de Sjögren , Humanos , Proteínas con Dominio MARVEL/metabolismo , Proteínas de la Mielina/metabolismo , Glándulas Salivales/metabolismoRESUMEN
Vascular cell adhesion molecule-1 (VCAM-1) and its ligand very late antigen (VLA-4) play important roles in many autoimmune diseases. Our study aimed to investigate the serum level of VCAM-1 and VLA-4 expression on peripheral blood neutrophil surface in patients with dermatomyositis (DM), especially focusing on patients with interstitial lung disease (ILD). Blood specimens of 42 patients with DM and 42 healthy controls matched for age and gender were recruited. Total serum VCAM-1 level was measured using commercial enzyme-linked immunosorbent assay (ELISA) and the percentages of VLA-4 expression on neutrophils were analyzed by flow cytometry. We divided patients into subgroups according to whether they had ILD and whether they exhibited diffuse alveolar damage (DAD) via high-resolution computed tomography (HRCT). sVCAM-1 was increased in classical DM (cDM) and clinical amyopathic dermatomyositis (CADM) compared with healthy controls (both p < .01). DM-ILD had higher sVCAM-1 levels than the none-ILD group (p < .01). sVCAM-1 was also significantly increased in the DAD group compared to the none-DAD group (p < .01). The percentages of VLA-4 expression on neutrophils in cDM and CADM patients were significantly elevated than that in healthy controls (both p < .01). The percentage of VLA-4 expression on neutrophils in DM patients with ILD was higher than none-ILD group (p < .01). In the patients with ILD, DAD group had a higher percentage of VLA-4 expression on neutrophils than none-DAD group (p < .01). Our findings indicated that serum VCAM-1 levels combined with VLA-4 expression on neutrophils might be useful for detecting the severity of lung disease in patients with DM.
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Dermatomiositis , Integrina alfa4beta1 , Enfermedades Pulmonares Intersticiales , Molécula 1 de Adhesión Celular Vascular , Humanos , Integrina alfa4beta1/metabolismo , Enfermedades Pulmonares Intersticiales/diagnóstico , Neutrófilos , Pronóstico , Estudios Retrospectivos , Molécula 1 de Adhesión Celular Vascular/sangreRESUMEN
Background: Recently trained immunity of microglia provided an opportunity to study the chronic effect of microglial activation and its metabolic rewiring in neuroimmunological diseases. Since elevated levels of B cell-activating factor (BAFF) have been proved to be associated with some chronic neuroimmunological disorders. Here, we used the trained innate immunity model to analyze the effect of BAFF, a vital regulator of the adaptive immune system, on long-term microglial activation and metabolic reprogramming in vitro and in vivo. Methods and results: In vitro, BV2 cells and mouse primary microglial cells were incubated with BAFF for 24 h (BAFF priming). After 5 days of resting, microglia were restimulated with LPS (LPS restimulation) or BAFF (BAFF restimulation). BAFF priming induced a pro-inflammatory trained immunity-phenotype of both BV2 cells and primary microglial cells, which was indicated by morphological change, secretion of pro-inflammatory cytokine and chemokine upon LPS restimulation or BAFF restimulation. The production of lactate and NAD+/NADH ratio were elevated 5 days after BAFF priming. The activation of the Akt/mTOR/HIF-1α pathway was induced by BAFF priming and lasted for 5 days. Pretreating the BV2 cells or mouse primary microglial cells with rapamycin blocked mTOR/HIF-1α activation and cellular metabolic reprogramming induced by BAFF training. Consistently, rapamycin efficiently suppressed the trained immunity-like responses of microglia triggered by BAFF. In vivo, adult male mice were treated with BAFF by intracerebroventricular injection for priming and 7 days later with BAFF for restimulation. BAFF training activated microglia in the cortex and hippocampus. The production of proinflammatory cytokines and chemokines was elevated after BAFF training. Conclusion: Our current data, for the first time, demonstrate that BAFF priming induces a proinflammatory memory-like response of microglia not only to LPS but also to BAFF itself. Rapamycin inhibits microglial priming triggered by BAFF through targeting the mTOR/HIF-1α signaling pathway. Our data reveal a novel role of BAFF in trained immunity and that rapamycin may be a potential therapeutic target of neuroimmunological diseases.
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Factor Activador de Células B/inmunología , Memoria Inmunológica/inmunología , Inmunosupresores/farmacología , Microglía/inmunología , Sirolimus/farmacología , Animales , Memoria Inmunológica/efectos de los fármacos , Inflamación/inmunología , Ratones , Microglía/efectos de los fármacosRESUMEN
Peripheral helper T (Tph) cells, phenotypically PD-1hiCXCR5-CD4+, are a recently identified Th cell subset that relates to several autoimmune diseases. Contrary to PD-1hiCXCR5+CD4+ follicular helper T (Tfh) cells, Tph cells are not located in lymphoid organs but accumulate in inflamed tissues. This study investigated Tph cells to determine their involvement in dermatomyositis (DM). The frequency of circulating Tph and Tfh cells was evaluated by flow cytometry at baseline and after glucocorticoid treatment. The expression of Tph and B cells was determined in muscle tissue by immunohistochemistry (IHC). Further, the correlations between circulating Tph cells and clinical characteristics were investigated. Flow cytometry revealed that circulating Tph and Tfh cells were decreased in peripheral blood of DM patients compared with healthy controls (HCs). However, the muscular expression of Tph and B cells was upregulated in patients with DM compared to that in the controls by IHC. Interestingly, the increased B cells accumulated around Tph cells in infiltrated lesions. The frequency of circulating Tph cells was positively correlated with Tfh cells, CD3+ T cells, CD4+ T cells, and CD8+ T cells, whereas negatively correlated with erythrocyte sedimentation rate (ESR), interleukin (IL)-6, and IL-10 levels. Furthermore, the abnormal circulating Tph cells in peripheral blood were recovered after glucocorticoid treatment. These results indicate that Tph cells might be involved in the immunopathogenesis of DM and therefore might provide novel insight for the development of DM therapies.
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Dermatomiositis , Linfocitos B , Linfocitos T CD4-Positivos , Humanos , Interleucinas , Músculos , Linfocitos T Colaboradores-InductoresRESUMEN
INTRODUCTION/OBJECTIVE: Ferritin has gained increasing attention in systemic lupus erythematosus (SLE).This study aimed to investigate the clinical significance of urinary ferritin/creatinine ratio in lupus nephritis (LN). METHOD: Samples from 62 SLE patients (35 with LN and 27 without LN) and 62 healthy controls were evaluated. There were nine patients who underwent renal biopsy. The amount of urinary ferritin was measured by enzyme-linked immunosorbent assay (ELISA) and normalized by the amount of urinary creatinine to obtain the urinary ferritin/creatinine ratio (UFCR). The relationships between UFCR and inflammatory markers, laboratory indicators, as well as the activity index (AI), and chronicity index (CI) of KBs were also investigated by correlation analysis. RESULTS: UFCR level in severe active SLE patients was significantly higher than that in inactive SLE patients (P < 0.01) or healthy controls (P < 0.01). In addition, UFCR level was significantly increased in the patients with LN when compared with those without LN (P < 0.01).Correlation analysis showed that UFCR level is positively correlated with Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) (P < 0.01), 24 h urine protein quantitation, serum creatinine, serum cystatin C, and GFR. The UFCR levels was significantly positively correlated with AI (P < 0.05) but not CI (P = 0.614) of KBs. CONCLUSIONS: UFCR level is a potential biomarker for the kidney injury in LN. Key Points ⢠UFCR level is significantly increased in LN patients. ⢠UFCR level is positively correlated with SLEDAI. ⢠UFCR level is closely related to kidney injury indicators. ⢠UFCR level is a potential biomarker for the kidney injury in LN.
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Lupus Eritematoso Sistémico , Nefritis Lúpica , Biomarcadores , Creatinina , Ferritinas , Humanos , Lupus Eritematoso Sistémico/diagnóstico , Nefritis Lúpica/diagnóstico , Índice de Severidad de la EnfermedadRESUMEN
BACKGROUND: Neuropsychiatric manifestations are frequent in patients with systemic lupus erythematosus (SLE), yet the etiology and pathogenesis of brain damage in SLE remains unclear. Because the production of autoantibodies, formation and deposition of immunocomplexes are major serological characteristics of SLE, the elevated level of serum immunoglobulin may contribute to brain tissue injury of SLE. To testify this, in this study, we examined whether immunoglobulin G (IgG) in the serum of SLE patients affects the cellular functions in central nervous system and the potential mechanism. METHODS: In vivo intracerebral injection of SLE-serum in mouse was used to activate microglia and the production of pro-inflammatory cytokine was assessed by ELISA. Sera was divided into IgG and IgG depleted fractions, while IgG was further divided into Fc and Fab fragments to examine which part has an effect on microglia. Flow cytometry, immunofluorescence and quantitative PCR (qPCR) were used to verify the synergistic effect of B-cell activating factor (BAFF) on IgG stimulation of microglia. RESULTS: We found that IgG in lupus sera can induce M1 activation of brain microglia following intraventricular injection into normal mice, and BAFF facilitates this process. In vitro, we identified that IgG bound to microglia through Fc rather than Fab fragments, and BAFF up-regulated the expression of Fc receptors (FcγR) on the surface of microglia, consequently, promote IgG binding to microglia. CONCLUSION: Our results suggest that lupus serum IgG causes inflammatory responses of microglia by involving the Fc signaling pathway and the activity could be up-regulated by BAFF. Accordingly, disruption of the FcγR-mediated signaling pathway and blockade of microglia activation may be a therapeutic target in patients with neuropsychiatric lupus erythematosus.
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Factor Activador de Células B/metabolismo , Fragmentos Fc de Inmunoglobulinas/metabolismo , Inmunoglobulina G/sangre , Lupus Eritematoso Sistémico/sangre , Microglía/patología , Animales , Línea Celular , Polaridad Celular , Citocinas/metabolismo , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Masculino , Ratones Endogámicos C57BL , Unión Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de IgG/metabolismo , Regulación hacia ArribaRESUMEN
OBJECTIVE: Brain-derived neurotrophic factor (BDNF) has been reported to be involved in the pathogenesis of autoimmune diseases and tyrosine kinase B (TrkB) is the specific receptor for BDNF. Our aim in this study was to investigate serum BDNF level and TrkB expression on peripheral blood T cell surface in patients with systemic lupus erythematosus (SLE) and explore potential relationship between serum BDNF and SLE. METHODS: Samples from fifty SLE patients and thirty healthy controls were evaluated. Serum BDNF level was measured by enzyme-linked immunosorbent assay (ELISA) and the percentages of TrkB expression on the surface of CD3â¯+â¯CD4â¯+â¯and CD3â¯+â¯CD8â¯+â¯T lymphocytes were measured by flow cytometry. The SLE patients were divided into subgroups according to whether they exhibited brain, kidney or lung involvement, and whether the disease was active or inactive. RESULTS: Serum BDNF levels in SLE patients were decreased when compared to the controls (pâ¯<â¯0.001). Comparing with the SLE individuals without systemic involvement, the BDNF levels were decreased in SLE patients with lupus nephritis (pâ¯=â¯0.042) and in SLE patients with neuropsychiatric manifestations (pâ¯=â¯0.04). On the other hand, the BDNF level was significantly increased in the inactive SLE group (pâ¯<â¯0.001) compared to the active SLE group. In addition, the percentages of TrkB expression on CD3â¯+â¯CD4â¯+â¯and CD3â¯+â¯CD8â¯+â¯T cell surface in SLE were significantly higher (pâ¯<â¯0.001; pâ¯<â¯0.001, respectively) than that in the controls. CONCLUSIONS: Serum BDNF level combined with TrkB expression on T cell surface can reflect SLE activity. It is possible that BDNF may be used as a potential serological biomarker for disease activity of SLE. In addition, the significant decrease in serum BDNF level may imply systemic involvement of SLE, as well as, possibly, differentiate neuropsychiatric SLE from hormone-induced mental disorders.
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Factor Neurotrófico Derivado del Encéfalo/biosíntesis , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/metabolismo , Regulación de la Expresión Génica , Lupus Eritematoso Sistémico/metabolismo , Glicoproteínas de Membrana/biosíntesis , Receptor trkB/biosíntesis , Adolescente , Adulto , Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/patología , Femenino , Humanos , Lupus Eritematoso Sistémico/patología , Masculino , Persona de Mediana EdadRESUMEN
Follicular helper T(Tfh) cells and follicular regulatory T(Tfr) cells are critical for the development and maintenance of germinal center and humoral immune responses. Accumulating evidence has demonstrated that the dysregulation of either Tfh or Tfr cells contributes to the pathogenesis of autoimmune diseases. The aim of this study was to examine the numbers of Tfh and Tfr cells in patients with rheumatoid arthritis (RA). Twenty-four patients with RA patients and 20 health controls (HCs) were enrolled in this study. We analyzed the numbers of Tfh (CD4+ CXCR5+ PD-1hi) cells and Tfr (CD4+ CXCR5+CD127lo) cells in 24 RA patients via flow cytometry. The level of the soluble PD-1 and its ligands (sPD-L1 and sPDL-2) were examined by ELISA. Flow cytometry revealed that both circulating Tfh and Tfr cells were increased in RA patients compared with HCs. More importantly, the ratio of Tfr/Tfh was decreased, indicating a disruption of the balance between Tfh and Tfr. The Tfr/Tfh ratio was inversely correlated with level of serum CRP, ESR, RF, anti-CCP, IgG and DAS28 index. We also found that the serum level of sPD-1 was significantly elevated in the RA patients, which was positively correlated with CRP, ESR and the number of Tfh cells. These results indicate that an imbalance of circulating Tfr and Tfh cells may be involved in the immunopathogenesis of RA and may provide novel insight for the development of RA therapies.
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Artritis Reumatoide/patología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunología , Adulto , Anciano , Antígeno B7-H1/sangre , Antígenos CD4/análisis , Células , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Humanos , Inmunofenotipificación , Subunidad alfa del Receptor de Interleucina-7/análisis , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Proteína 2 Ligando de Muerte Celular Programada 1/sangre , Receptor de Muerte Celular Programada 1/análisis , Receptores CXCR5/análisisRESUMEN
PURPOSE: The resistance to 5-FU often limits its clinical effectiveness on breast cancer treatment. Combination therapy thus is employed to overcome this treatment resistance. We here report a potent antitumor effect of Emodin at low dose on chemotherapy sensitivity of MCF-7 breast cancer cells. METHODS: Cell viability, apoptosis, glutathiones (GSH) concentration and Reactive oxygen species (ROS) activity following Emodin and 5-FU treatment was assessed. Cellular senescence following combined treatment and silence of NRARP was examined by senescence-associated ß-galactosidase analysis. Western blot analysis was used to determine changes in the expression of p21, p16, p27, E2F1 and NRARP. RESULTS: Low dose Emodin potentiates 5-FU-induced apoptosis of breast cancer cells, in association with inhibition of NRARP, resulting in cellular senescence. RNA interference of NRARP induced cellular senescence in MCF-7 breast cancer cells. Furthermore, the cellular senescence induced by Emodin and 5-FU treatment could be reverted by pcDNA-NRARP. CONCLUSION: These findings provide preclinical evidence for repurposing use of Emodin in combination with chemotherapeutic agents to treat breast cancer as an alternative salvage regimen.
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Antimetabolitos Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Emodina/farmacología , Fluorouracilo/farmacología , Proteínas de Neoplasias/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular , Células MCF-7 , Proteínas de Neoplasias/metabolismo , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
OBJECTIVE: To examine the expression of chemokine receptors in different peripheral blood T-cell subsets in patients with polymyositis (PM) and dermatomyositis (DM). METHODS: We used flow cytometry to measure the frequencies of chemokinereceptors CXCR3 and CCR4 expression in the CD4+ or CD8+ lymphocytes. Enzyme linked immunosorbent assays were also used to measure the concentrations of C-X-C motif chemokine 10 (CXCL10), thymus and activation regulated chemokine (TARC) and macrophage derived chemokine (MDC). RESULTS: Comparing to 20 healthy controls, %CD4+CXCR3+ and %CD8+CXCR3+ T cells significantly decreased in 33DM patients, and %CD8+CXCR3+ cells decreased in 24PM patients, but %CD4+CCR4+ and %CD8+CCR4+ cells did not significantly change in both the PM and DM patients. Accordingly, the Th1/Th2 polarization, analyzed as the balance obtained after dividing %CD4+CXCR3+ cells by %CD4+CCR4+ cells, showed a significant reduction in DM. The serum concentration of CXCR3+ ligand, CXCL10, significantly increased and negatively correlated with circulating %CD4+CXCR3+ cells in DM patients. There was no significant change of TARC and MDC in PM and DM patients. Furthermore, %CD4+CXCR3+ cells decreased more severely in the patients with interstitial lung disease. CONCLUSIONS: The present results indicate that the distributions of circulating CXCR3+ T-cells differ among the PM and DM cases. Our findings suggest a pathogenic difference between PM and DM.
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Dermatomiositis/sangre , Polimiositis/sangre , Receptores de Quimiocina/sangre , Adolescente , Adulto , Anciano , Estudios de Casos y Controles , Humanos , Ligandos , Persona de Mediana Edad , Adulto JovenRESUMEN
To investigate the potential involvement of microglia in the neuropathology of systemic lupus erythematosus (SLE), we examined whether SLE patient sera could activate BV2 microglia in vitro. Exposure to SLE patient sera resulted in morphological changes in the microglia, an increase in MHC II and CD86 protein expression, and an obvious release of nitric oxide and proinflammatory cytokines. However, the SLE sera did not induce a specific change in the production of immunoregulatory cytokines. Inactivating complements or neutralizing proinflammatory cytokines in the SLE sera did not suppress microglial activation. Our results highlight the potential role of microglia in neuroinflammation in SLE patients.
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Lupus Eritematoso Sistémico/sangre , Microglía/metabolismo , Suero/metabolismo , Adulto , Animales , Anticuerpos/farmacología , Estudios de Casos y Controles , Moléculas de Adhesión Celular/metabolismo , Línea Celular Transformada , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Medio de Cultivo Libre de Suero/farmacología , Citocinas/metabolismo , Femenino , Antígenos de Histocompatibilidad Clase I/inmunología , Antígenos de Histocompatibilidad Clase I/metabolismo , Humanos , Masculino , Ratones , Microglía/efectos de los fármacos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Estudios RetrospectivosRESUMEN
DNase1l3 is an endonuclease to degrade the chromatin of apoptotic or necrotic cells. Serum DNase1l3 may fulfill the function of clearance of chromatin released into the circulation by dying cells, which can trigger autoimmune responses. To date, it remains unclear whether serum DNase1l3 level associates with the pathogenesis of autoimmune diseases. Sixty-eight patients with dermatomyositis/polymyositis (DM/PM, n = 30), systemic lupus erythematosus (SLE, n = 20) and rheumatoid arthritis (RA, n = 18), as well as 26 healthy blood donors were enrolled in the present study. Serum levels of DNase1l3 were quantified by enzyme-linked immunosorbent assay. DNASE1L3 activity in serum was estimated by the capability of serum to digest nucleosomal DNA. Clinical, biochemical, serological and other markers of disease activity (CRP, ESR, C3, C4, anti-Jo-1 and anti-dsDNA, etc.) were measured by standard laboratory procedure. We found a decrease in DNase1l3 level in the DM/PM and SLE patients, resulting in the reduction in serum activity to digest nucleosome DNA. In contrast, the level and activity of DNase1l3 remained unchanged in the RA patients. The DNase1l3 level was relatively lower in the DM/PM patients with anti-Jo-1 antibody and interstitial lung disease, and in the SLE patients with SLE disease activity index higher than 6, renal involvement and anti-dsDNA antibody. DNase1l3 level negatively correlated with CRP and IgG in the PM/DM patients and correlated with ESR in the SLE patients. We found a significant reduction in serum DNase1l3 level in DM/PM and SLE, which may associate with clinic features and disease activity.
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Artritis Reumatoide/patología , Dermatomiositis/patología , Endodesoxirribonucleasas/sangre , Lupus Eritematoso Sistémico/patología , Adolescente , Adulto , Anciano , ADN/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Hidrólisis , Masculino , Persona de Mediana Edad , Suero/química , Adulto JovenRESUMEN
OBJECTIVE: Although chemokines are critical elements for the selective attraction and activation of various leukocyte subsets in the inflammatory process, there are few findings concerning T helper (Th) 1 or Th2 chemokines in ankylosing spondylitis (AS). This study was designed to determine whether serum levels of chemokines that are preferentially chemotactic for Th1 (IFN-gamma-inducible protein-10, IP-10/CXCL10) and Th2 (thymus and activation regulated chemokine, TARC/CCL17) and (macrophage derived chemokine, MDC/CCL22) cells were elevated and whether they correlated with the clinical features in patients with AS. METHODS: Forty-two patients with axial AS and 25 healthy controls were enrolled into the study. Serum levels of chemokines (IP-10, TARC and MDC) and cytokines (IFN-γ, TNF-α and IL-4) were examined using ELISA. The disease activity was evaluated by Ankylosing Spondylitis Disease Activity Score (ASDAS). Serum levels of IgG, IgA, IgM, erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were measured. RESULTS: Serum chemokine levels of IP-10, TARC and MDC were significantly higher in patients with AS than those in healthy controls. Serum cytokine levels of IFN-γ, TNF-α were also significantly increased, but the levels of IL-4 were not. Furthermore, IP-10 levels in AS patients correlated with ESP, CRP and ASDAS, while the levels of TARC and MDC did not correlate with these clinic indexes. Correlation analysis between the levels of chemokines and cytokines revealed a positive correlation between IP-10 and TNF-α. The levels of both Th1 and Th2 chemokines decreased under blockade of TNF-α. CONCLUSION: Our results suggest that both a Th1 chemoattractant IP-10 and Th2 chemoattractants, TARC and MDC, cooperatively play a role in the development of AS.
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Quimiocina CCL17/inmunología , Quimiocina CCL22/metabolismo , Quimiocina CXCL10/metabolismo , Espondilitis Anquilosante/metabolismo , Células TH1/metabolismo , Células Th2/metabolismo , Adulto , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Quimiocina CCL17/sangre , Quimiocina CCL22/sangre , Quimiocina CXCL10/sangre , Citocinas/sangre , Citocinas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina M/sangre , Mediadores de Inflamación/sangre , Mediadores de Inflamación/metabolismo , Masculino , Espondilitis Anquilosante/sangre , Espondilitis Anquilosante/inmunología , Adulto JovenRESUMEN
The aim of this study was to investigate the effects of emodin on the proliferation of human breast cancer cells Bcap-37 and ZR-75-30. Cell viability following emodin treatment was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of emodin on apoptosis were determined by flow cytometry using Annexin V-fluorescein isothiocyanate and propidium iodide staining. Quantitative polymerase chain reaction and western blot analysis were used to determine changes in the expression of apoptotic genes and protein, respectively. The effect of emodin on the invasiveness of breast cancer cells was evaluated by Matrigel invasion assay. Treatment of breast cancer cells Bcap-37 and ZR-75-30 with emodin was observed to inhibit the growth and induced apoptosis in a time- and dose-dependent manner. Emodin reduced the level of Bcl-2 and increased levels of cleaved caspase-3, PARP, p53 and Bax. These findings indicate that emodin induces growth inhibition and apoptosis in human breast cancer cells. Emodin may be a potential therapeutic agent for the treatment of breast cancer.
RESUMEN
In the United States of America, prostate cancer is the second most common age-related cancer among men. African-American men have the highest incidence of, and mortality rate from this disease in the United States. According to the American Cancer Society, 29% of all cancer cases and 9% of all cancer deaths are a result of prostate cancer. Individuals who are at highest risk include African-American men, men over 60 years of age, and those with a family history of the disease. African-Americans also have twice the risk of developing prostate cancer as compared to Caucasians. Erythroblastosis virus E26 transformation-specific (ETS) factors play an important role in human cancers. ETS Variant 1 (ETV1), an ETS factor, is notable for its association in prostate cancers, where truncated ETV1 (dETV1) or its full length counterpart is overexpressed in approximately 10% of the prostate cancer patients. Prostate cancer tumorigenesis may be initiated by deregulation of the Wnt/ß-catenin pathway. Mutations that stabilize ß-catenin were shown to contribute to the loss of cell-growth control in tumorigenesis. We hypothesized that ETV1's interaction with components of the Wnt/ß-catenin pathway may alter ß-catenin's interaction with downstream tumor-suppressor genes, which are critical in regulating apoptosis and cell-growth properties of prostate cells. Our results demonstrate for the first time that ETV1 alters ß-catenin activity by activating kinases that regulate Wnt/ß-catenin activity through post-translational modification in prostate cancer cells. We further demonstrate that therapeutic agents such as PD98059, that reverse effect of ETV1 on Wnt/ß-catenin signaling pathway, can be used to target ETV1-positive prostate cancer cells. These therapeutic agents could have a profound impact on prevention and treatment of prostate cancer which may help to reduce health disparity seen in minority patients. Understanding the role of ETV1 in Wnt/ß-catenin pathway will also allow us to develop better diagnostic tools, which can be used as a biomarker for ETV1-positive prostate cancers.
RESUMEN
The aim of this study is to investigate the serum levels and clinical significance of nerve grow factor (NGF) and brain-derived neurotrophic factor (BDNF) in Sjogren's syndrome (SS) with interstitial lung disease (ILD). Fifty two untreated patients with SS were enrolled in the study. Of them, 25 patients only displayed salivary glands damage and/or lacrimal gland injury (simple SS group). The other 27 patients were lacrimal and/or salivary gland involvement as well as being concomitant only with intestinal lung disease (ILD group). Twenty-five serum samples from healthy volunteers were examined as controls. We measure serum NGF and BDNF levels by ELISA and correlate them with clinical data. Serum NGF levels were significantly higher in ILD patients (372 ± 129 pg/ml) and simple SS patients (293 ± 72 pg/ml) when compared with healthy controls (187 ± 47 pg/ml) (both p < 0.01). Significant difference were also found between the two patient groups (p < 0.01). In contrast, BDNF were significantly decreased in ILD patients (1,005 ± 143 pg/ml) when compared with either simple SS patients (1,204 ± 176 pg/ml, p < 0.01) or healthy controls (1,217 ± 155 pg/ml, p < 0.01). Correlation analysis showed NGF levels in ILD patient were positively correlated with serum levels of C-reactive protein and IgG (both p < 0.05). The abnormal NGF and BDNF in sera may be a potential character of ILD secondary to pSS.
Asunto(s)
Factor Neurotrófico Derivado del Encéfalo/sangre , Enfermedades Pulmonares Intersticiales/sangre , Factor de Crecimiento Nervioso/sangre , Síndrome de Sjögren/sangre , Adulto , Anciano , Femenino , Humanos , Enfermedades Pulmonares Intersticiales/complicaciones , Masculino , Persona de Mediana Edad , Síndrome de Sjögren/complicacionesRESUMEN
Transforming growth factor beta (TGF-ß) signaling pathway is involved in diverse cellular processes, including cell proliferation, differentiation, adhesion, apoptosis, and some human diseases including cancer. Smad proteins function as mediators of intracellular signal transduction of TGF-ß. Following their phosphorylation by TGF-ß receptor I, Smad2 and Smad3 form a heteromeric complex with Smad4 and then are translocated into the nucleus where they bind to other co-factors and regulate the expression of target genes. ERG (Ets Related Gene) belongs to the ETS family of transcriptional factors. Chromosomal rearrangement of TMPRSS2 gene and ERG gene has been found in the majority of prostate cancers. Over-expression of full length or truncated ERG proteins is associated with a higher rate of recurrence and unfavorable prognosis. In this review, we focus on recent understanding of regulation of TGF-ß/Smads signaling pathway by ERG proteins in prostate cancer.
RESUMEN
TGF-ß/Smads signaling plays a significant role in the regulation of growth of normal and prostate cancer cells. Smad proteins function as important mediators of intracellular signal transduction of transforming growth factor-ß (TGF-ß). TGF-ß signaling pathway is known to regulate cell proliferation, differentiation, apoptosis and play a major role in some human diseases and cancers. Following their phosphorylation by TGF-ß receptor-I, Receptor-regulated Smads (including Smad2 and Smad3 proteins) form a heteromeric complex with co-Smad (Smad4) and then translocate into the nucleus where they bind and regulate the expression of target genes. ERG (Ets Related Gene) belongs to the ETS family of transcriptional factors. Chromosomal rearrangement of TMPRSS2 gene and ERG gene has been found in majority of prostate cancers. Over-expression of full length or truncated ERG proteins have been shown to associate with a higher rate of recurrent and unfavorable prognosis of prostate cancer. In order to understand how ERG oncoprotein regulates TGF-ß/Smads signaling pathway, we have studied the effect of ERG on TGF-ß/Smad3 signaling pathway. In this study, we demonstrate that ERG oncoprotein physically interacts with Smad3 protein and stabilizes phospho-Smad3 protein and thereby enhance TGF-ß/Smad3 signaling pathway in prostate cells. Thus, ERG oncoprotein plays an important role in prostate tumorigenesis by using a novel mechanism to activate TGF-ß/Smad3 signaling pathway.