transforming-growth-factor-beta and Lupus-Erythematosus--Systemic

The helper T cell 9 (Thelper-9, Th9), as a functional subgroup of CD4

Topics: Animals; Cell Differentiation; GATA3 Transcription Factor; Gene Expression Regulation; Humans; Inflammation; Interleukin-4; Interleukin-9; Lupus Erythematosus, Systemic; Mice; Mice, Knockout; Neoplasms; Proto-Oncogene Proteins; Signal Transduction; STAT6 Transcription Factor; T-Lymphocytes, Helper-Inducer; Trans-Activators; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a chronic, systemic, autoimmune inflammatory disease characterized by the production of numerous autoantibodies and cytokines, as well as multiple organ damage. Specific B cell subsets negatively regulate immune responses and have been termed regulatory B cells (Bregs). Bregs are characterized by the production of the immunoregulatory cytokines interleukin (IL)-10, IL-35, and transforming growth factor (TGF)-

Topics: Autoimmune Diseases; B-Lymphocytes, Regulatory; Cytokines; Humans; Interleukin-10; Interleukins; Lupus Erythematosus, Systemic; Phenotype; Polymorphism, Genetic; Receptors, Interleukin-10; Transforming Growth Factor beta

Transforming growth factor β (TGF-β) is a pleiotropic cytokine involved in both suppressive and inflammatory immune responses. After 30 years of intense study, we have only begun to elucidate how TGF-β alters immunity under various conditions. Under steady-state conditions, TGF-β regulates thymic T-cell selection and maintains homeostasis of the naïve T-cell pool. TGF-β inhibits cytotoxic T lymphocyte (CTL), Th1-, and Th2-cell differentiation while promoting peripheral (p)Treg-, Th17-, Th9-, and Tfh-cell generation, and T-cell tissue residence in response to immune challenges. Similarly, TGF-β controls the proliferation, survival, activation, and differentiation of B cells, as well as the development and functions of innate cells, including natural killer (NK) cells, macrophages, dendritic cells, and granulocytes. Collectively, TGF-β plays a pivotal role in maintaining peripheral tolerance against self- and innocuous antigens, such as food, commensal bacteria, and fetal alloantigens, and in controlling immune responses to pathogens.

Topics: Animals; Arthritis, Rheumatoid; Autoimmunity; B-Lymphocytes; Bacterial Infections; Cell Differentiation; Cell Lineage; Cell Proliferation; Cell Survival; Dendritic Cells; Diabetes Mellitus, Type 1; Granulocytes; Homeostasis; Humans; Immune Tolerance; Inflammatory Bowel Diseases; Isoantigens; Killer Cells, Natural; Lupus Erythematosus, Systemic; Lymphocyte Activation; Macrophages; Mast Cells; Mice; Monocytes; Parasitic Diseases; T-Lymphocytes; T-Lymphocytes, Regulatory; Thymus Gland; Transforming Growth Factor beta

SLE is an autoimmune disease characterized by multiple organ damage mediated by autoantibodies and autoreactive T cells. Approaches utilizing genetically engineered mice as well as genome-wide association studies have identified a number of lupus-related genes. Recently, early growth response gene 2 (Egr2) and Egr3 have emerged as regulatory molecules that suppress excessive immune responses. Mice deficient for Egr2 and Egr3 develop a lupus-like disease with dysregulated activation of effector T cells. Furthermore, Egr2 and Egr3 confer suppressive activity to CD4

Topics: Animals; Autoimmune Diseases; Autoimmunity; CD4-Positive T-Lymphocytes; Early Growth Response Protein 2; Early Growth Response Protein 3; Humans; Interleukin-10; Interleukin-17; Lupus Erythematosus, Systemic; Mice; T-Lymphocytes; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease with different variety of clinical manifestations. Natural killer T (NKT) cells are innate lymphocytes that play a regulatory role during broad range of immune responses. A number of studies demonstrated that the quantity and quality of invariant NKT (iNKT) cells showed marked defects in SLE patients in comparison to healthy controls. This finding suggests that iNKT cells may play a regulatory role in the occurrence and development of this disease. In this review, we mainly summarized the most recent findings about the behavior of NKT cells in SLE patients and mouse models, as well as how NKT cells affect the proportion of T helper cells and the production of autoreactive antibodies in the progress of SLE. This will help people better understand the role of NKT cells in the development of SLE and improve the therapy strategy.

Topics: Animals; Autoantibodies; Disease Models, Animal; Female; Gene Expression; Humans; Immunosuppressive Agents; Interleukin-10; Interleukin-17; Lupus Erythematosus, Systemic; Mice; Natural Killer T-Cells; Survival Analysis; Th1 Cells; Th1-Th2 Balance; Th2 Cells; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disorder that disproportionally affects women, especially in their reproductive years. SLE is associated with considerable pregnancy-related morbidity--including fetal loss, preterm birth, fetal growth restriction and pre-eclampsia. CD4+CD25+ regulatory T (T(REG)) cells have a potent immunosuppressive function and contribute to immunological self-tolerance. These cells might be essential for successful placental development by ensuring fetal tolerance. The numbers of T(REG) cells are augmented during normal pregnancy and, conversely, diminished numbers are associated with pregnancy loss and pre-eclampsia. Several studies have shown that patients with SLE have decreased numbers of T(REG) cells that might be functionally defective. This defective T(REG) cell functioning could predispose women with SLE to pregnancy complications. This article provides an overview of current knowledge of the role and function of T(REG) cells in SLE and pregnancy and how these cells might contribute to improving pregnancy-related outcomes in patients with SLE in the future.

Topics: Female; Humans; Lupus Erythematosus, Systemic; Pregnancy; Pregnancy Complications; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production, complement activation, and immune complex deposition, resulting in tissue and organ damage. An understanding of the mechanisms responsible for homeostatic control of inflammation, which involve both innate and adoptive immune responses, will enable the development of novel therapies for SLE. Regulatory T cells (Treg) play critical roles in the induction of peripheral tolerance to self- and foreign antigens. Naturally occurring CD4+CD25+ Treg, which characteristically express the transcription factor forkhead box protein P3 (Foxp3), have been intensively studied because their deficiency abrogates self-tolerance and causes autoimmune disease. Moreover, regulatory cytokines such as interleukin-10 (IL-10) also play a central role in controlling inflammatory processes. This paper focuses on Tregs and Treg-associated cytokines which might regulate the pathogenesis of SLE and, hence, have clinical applications.

Topics: Autoimmune Diseases; Humans; Interleukin-10; Interleukin-17; Interleukins; Lupus Erythematosus, Systemic; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

The emerging role of interleukin (IL)-17 as a hallmark proinflammatory cytokine of the adaptive immune system, produced primarily by a new T helper cell subset termed 'Th17', has received considerable attention. Differentiation of Th17 cells is driven by the simultaneous presence of transforming growth factor-beta and certain inflammatory cytokines (e.g. IL-6, IL-21), and recent studies have shown that inflammation instigated by IL-17-producing cells is central to the development and pathogenesis of several human autoimmune diseases and animal models of autoimmunity. In this review, we focus on the information regarding IL-17 and systemic lupus erythematosus (SLE), a chronic autoimmune disease. The work that has explored the development and behaviour of IL-17-producing cells in SLE is discussed, and different mechanisms by which IL-17 could potentially augment inflammation and autoantibody production in the context of SLE are proposed.

Topics: Animals; Autoantibodies; Humans; Interleukin-17; Lupus Erythematosus, Systemic; Mice; Mice, Mutant Strains; Models, Animal; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Regulatory T cells (Tregs) maintain immunological homeostasis and prevent autoimmunity. The depletion or functional alteration of Tregs may lead to the development of autoimmune diseases. Tregs consist of different subpopulations of cells, of which CD4(+)CD25(+)Foxp3(+) cells are the most well characterized. However, CD8 Tregs also constitute a major cell population that has been shown to play an important role in autoimmune diseases. This review will discuss the role of Tregs in autoimmune diseases in general and specifically in systemic lupus erythematosus (SLE). SLE is a multisystem autoimmune disease characterized by the production of autoantibodies against nuclear components and by the deposition of immune complexes in the kidneys as well as in other organs. Abnormalities in Tregs were reported in SLE patients and in animal models of the disease. Current treatment of SLE is based on immunosuppressive drugs that are nonspecific and may cause adverse effects. Therefore, the development of novel, specific, side effect-free therapeutic means that will induce functional Tregs is a most desirable goal. Our group and others have designed and utilized tolerogenic peptides that ameliorate SLE manifestations in murine models. Here, we demonstrate the role of CD4 and CD8 Tregs, as well as the interaction between the two subsets of cells and the mechanism of action of the tolerogenic peptides. We also discuss their therapeutic potential for the treatment of SLE.

Topics: Animals; Antibodies, Monoclonal; Antigen-Presenting Cells; Autoimmunity; Forkhead Transcription Factors; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Peptide Fragments; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

A central pathologic mechanism in systemic autoimmune diseases with chronic inflammation such as systemic lupus erythematosus (SLE) is the aberrant production of antibodies against self-components produced by abnormal B cells with the help of hyperactive CD4(+)T cells. One goal for better control of the disease is the limitation of the number of abnormal and hyperactive cells, to prevent and/or attenuate the damaging effects of the pathogenic antibodies on target tissues. Recently, a role of regulatory T cells in the suppression of autoimmune reactivity in diseases including SLE has been recognized. CD4(+)CD25(+) regulatory T cells (Tregs) and CD8(+) inhibitory T (Ti) cells have been found numerically decreased and/or functionally impaired in some patients with active systemic lupus erythematosus. Recent experimental work and preclinical studies have also provided proof-of-concept for the possibility of induction of self-tolerance through the modulation of regulatory/suppressor T cells using self antigen-derived peptides that could promote suppression of the production of pathogenic antibodies. This review explores the mechanisms elicited by the administration of self antigen-derived peptides on the induction of suppression of autoimmune responses, and how this information might lead to future development of new strategies for better management of systemic autoimmune conditions.

Topics: Animals; Autoimmune Diseases; CD28 Antigens; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Humans; Immune Tolerance; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Peptides; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

TGF-beta is a cytokine of great importance in many common diseases because it takes part in many physiological processes such as angiogenesis and stimulation of the synthesis and degradation of extracellular matrix proteins. It also regulates the entrance of cells to the apoptotic pathway and can stimulate the division of mesenchymal cells and inhibit hemopoietic, endothelial, and lymphatic cells. There are five genes which encode TGF-beta in vertebrates, of which only three are present in mammals. The best known member of the family of TGF-beta proteins is TGF-beta 1. TGF-beta is synthetized as a precursor protein which, after enzymatic modification, is present as a small or large complex. Three membrane receptors, serine/threonine kinase, are arranged for signal transduction with TGF-beta. Smad proteins are responsible for sending the signal into the cell nucleus; its influence on different transcriptive factors in the cell nucleus promotes the expressions of different genes. Disturbances in TGF-beta expression have been noted in many diseases. Current results clearly indicate an important role of this cytokine in autoimmunological disorders, especially in systemic lupus erythematosus. Studies on an animal model revealed that endogenic TGF-beta can control the progression of systemic lupus erythematosus.

Topics: Humans; Lupus Erythematosus, Systemic; Signal Transduction; Transforming Growth Factor beta

Naturally occurring CD4+CD25+ regulatory T cells mediate immune suppression to limit immunopathogenesis associated with chronic inflammation, persistent infections and autoimmune diseases. Their mode of suppression is contact-dependent, antigen-nonspecific and involves a nonredundant contribution from the cytokine transforming growth factor (TGF)-beta. Not only can TGF-beta mediate cell-cell suppression between the regulatory T cells and CD4+CD25- or CD8+ T cells, but new evidence also reveals its role in the conversion of CD4+CD25- T cells, together with TCR antigen stimulation, into the regulatory phenotype. Elemental to this conversion process is induction of expression of the forkhead transcription factor, Foxp3. This context-dependent coercion of naive CD4+ T cells into a powerful subset of regulatory cells provides a window into potential manipulation of these cells to orchestrate therapeutic intervention in diseases characterized by inadequate suppression, as well as a promising means of controlling pathologic situations in which excessive suppression dominates.

Topics: Animals; Arthritis, Rheumatoid; Asthma; Autoimmune Diseases; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Clonal Anergy; Disease Models, Animal; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Immune Tolerance; Immunotherapy, Adoptive; Inflammation; Lupus Erythematosus, Systemic; Mice; Mice, Knockout; Receptors, Interleukin-2; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Lupus is a chronic autoimmune inflammatory disease with complex clinical manifestations. In humans, lupus, also known as systemic lupus erythematosus (SLE), affects between 40 and 250 individuals, mostly females, in each 100 000 of the population. There are also a number of murine models of lupus widely used in studies of the genetics, immunopathology, and treatment of lupus. Human patients and murine models of lupus manifest a wide range of immunological abnormalities. The most pervasive of these are: (1) the ability to produce pathogenic autoantibodies; (2) lack of T- and B-lymphocyte regulation; and (3) defective clearance of autoantigens and immune complexes. This article briefly reviews immunological abnormalities and disease mechanisms characteristic of lupus autoimmunity and highlight recent studies on the use of gene therapy to target these abnormalities.

Topics: Abatacept; Animals; Antigen-Antibody Complex; Antigens, CD; Antigens, Differentiation; Autoantibodies; B-Lymphocytes; Complement System Proteins; CTLA-4 Antigen; Cytokines; Disease Models, Animal; Gene Transfer Techniques; Genetic Predisposition to Disease; Genetic Therapy; Humans; Immunoconjugates; Immunotherapy; Killer Cells, Natural; Lupus Erythematosus, Systemic; Mice; Phagocytosis; T-Lymphocytes; Transforming Growth Factor beta

Regulatory T cells prevent autoimmunity by suppressing the reactivity of potentially aggressive self-reactive T cells. Contact-dependent CD4+ CD25+ 'professional' suppressor cells and other cytokine-producing CD4+ and CD8+ T-cell subsets mediate this protective function. Evidence will be reviewed that T cells primed with transforming growth factor (TGF)-beta expand rapidly following restimulation. Certain CD4+ T cells become contact-dependent suppressor cells and other CD4+ and CD8+ cells become cytokine-producing regulatory cells. This effect is dependent upon a sufficient amount of IL-2 in the microenvironment to overcome the suppressive effects of TGF-beta. The adoptive transfer of these suppressor cells generated ex vivo can protect mice from developing chronic graft-versus-host disease with a lupus-like syndrome and alter the course of established disease. These data suggest that autologous T cells primed and expanded with TGF-beta have the potential to be used as a therapy for patients with systemic lupus erythematosus and other chronic inflammatory diseases. This novel adoptive immunotherapy also has the potential to prevent the rejection of allogeneic transplants.

Topics: Animals; CD4-Positive T-Lymphocytes; Cell Transplantation; Humans; Immunotherapy; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Receptors, Interleukin-2; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Topics: Animals; Arthritis, Experimental; Autoimmune Diseases; Cytokines; Encephalomyelitis, Autoimmune, Experimental; Immunotherapy; Interferon-gamma; Interleukins; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NOD; Mice, Mutant Strains; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Reports of clinical improvement following mesenchymal stromal cell (MSC) infusions in refractory lupus patients at a single centre in China led us to perform an explorative phase I trial of umbilical cord derived MSCs in patients refractory to 6 months of immunosuppressive therapy.. Of six patients, five (83.3%; 95% CI 35.9% to 99.6%) achieved the clinical endpoint of an SRI of 4. Adverse events were minimal. Mechanistic studies revealed significant reductions in CD27IgD double negative B cells, switched memory B cells and activated naïve B cells, with increased transitional B cells in the five patients who met the endpoint. There was a trend towards decreased autoantibody levels in specific patients. Two patients had increases in their Helios+Treg cells, but no other significant T cell changes were noted. GARP-TGFβ complexes were significantly increased following the MSC infusions. The B cell changes and the GARP-TGFβ increases significantly correlated with changes in SLEDAI scores.. This phase 1 trial suggests that umbilical cord (UC) MSC infusions are very safe and may have efficacy in lupus. The B cell and GARP-TGFβ changes provide novel insight into mechanisms by which MSCs may impact disease.. NCT03171194.

Topics: Female; Humans; Lupus Erythematosus, Systemic; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Transforming Growth Factor beta; Umbilical Cord

Topics: Animals; Brain-Derived Neurotrophic Factor; Calcitriol; Choroid Plexus; Inflammation; Lupus Erythematosus, Systemic; Mice; Mice, Inbred MRL lpr; NF-kappa B; PPAR gamma; Smad Proteins; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Despite advances in general and targeted immunosuppressive therapies, limiting all mainstay treatment options in refractory systemic lupus erythematosus (SLE) cases has necessitated the development of new therapeutic strategies. Mesenchymal stem cells (MSCs) have recently emerged with unique properties, including a solid propensity to reduce inflammation, exert immunomodulatory effects, and repair injured tissues.. An animal model of acquired SLE mice was induced via intraperitoneal immunization with Pristane and affirmed by measuring specific biomarkers. Bone marrow (BM) MSCs were isolated from healthy BALB/c mice and cultured in vitro, then were identified and confirmed by flow cytometry and cytodifferentiation. Systemic MSCs transplantation was performed and then several parameters were analyzed and compared, including specific cytokines (IL-17, IL-4, IFN-ɣ, TGF-β) at the serum level, the percentage of Th cell subsets (Treg/Th17, Th1/Th2) in splenocytes, and also the relief of lupus nephritis, respectively by enzyme-linked immunosorbent assay (ELISA), flow cytometry analysis and by hematoxylin & eosin staining and also immunofluorescence assessment. Experiments were carried out with different initiation treatment time points (early and late stages of disease). Analysis of variance (ANOVA) followed by post hoc Tukey's test was used for multiple comparisons.. The rate of proteinuria, anti-double-stranded deoxyribonucleic acid (anti-dsDNA) antibodies, and serum creatinine levels decreased with BM-MSCs transplantation. These results were associated with attenuated lupus renal pathology in terms of reducing IgG and C3 deposition and lymphocyte infiltration. Our findings suggested that TGF-β (associated with lupus microenvironment) can contribute to MSC-based immunotherapy by modulating the population of TCD4. MSC-based immunotherapy showed a delayed effect on the progression of acquired SLE in a lupus microenvironment-dependent manner. Allogenic MSCs transplantation revealed the ability to re-establish the balance of Th17/Treg, Th1/Th2 and restore the plasma cytokines network in a pattern dependent on disease conditions. The conflicting results of early versus advanced therapy suggest that MSCs may produce different effects depending on when they are administered and their activation status.

Topics: Animals; Cytokines; Lupus Erythematosus, Systemic; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a disorder with a complex immunopathogenesis. It is well known that the disease begins with immunological alterations and autoantibody appearance in the serum years before clinical onset. As SLE has a strong tendency to familial aggregation, first-degree relatives (FDRs) constitute a group at elevated risk. The current understanding is that external risk factors trigger underlying immune dysregulations, leading to overt disease in those with elevated genetic risk.. This cross-sectional study investigates the degree to which clinical features, external risk factors, and immunological profiles differ in SLE FDRs from healthy individuals and SLE patientts.. Three groups were studied: Lupus patient FDRs (n = 56), healthy controls (n = 20), and SLE patients (n = 20). FDRs and healthy participants completed a detailed clinical questionnaire that included questions regarding smoking and estrogen drug history. All participants were tested for the presence of the following antinuclear autoantibodies (ANAs) against: nRNP/Sm, Sm, Ro60, Ro-52, La, Scl-70, PM-Scl, PM- Scl, Jo-1, CENP B, PCNA, dsDNA, nucleosomes, histones, RibP, AMA M2, DFS70, and eight soluble cytokines, including transforming growth factor-β (TGF-β), vitamin D levels, and antibodies against Epstein-Barr virus (EBV).. Compared with the healthy controls, FDRs had higher titers of ANA, more specific staining immunofluorescent patterns, and more autoantibody specificities. Furthermore, FDRs differed significantly in their TGF-β levels from the other two groups. In FDRs, some clinical features (hair loss, skin, and oral ulcer-like lesions) were associated with higher ANA titers and some (oral ulcer-like lesions) with the anti-Ro60-specific antibody. Interestingly, there was an association between ANA titers and levels of antibodies against EBV only in the FDR group.. First-degree relatives display unique clinical and immunological profiles, placing them between healthy individuals and SLE patients, with a balance between compensated immune dysregulation and disease-developing potential. A possible association between ANA titer and the number of clinical complaints is observed, which needs to be confirmed in more extensive studies.

Topics: Antibodies, Antinuclear; Autoantibodies; Cross-Sectional Studies; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; Lupus Erythematosus, Systemic; Oral Ulcer; Transforming Growth Factor beta

Jieduquyuziyin prescription (JP) is a traditional Chinese medicine utilized to treat systemic lupus erythematosus (SLE). Its efficacy has been confirmed through clinical trials and empirical evidence, leading to its authorized use in Chinese hospitals. The development of JP exemplifies the integration of traditional wisdom and scientific approaches, demonstrating the interdisciplinary essence of ethnopharmacology. These results emphasize the potential value of traditional medicine in addressing autoimmune disorders.. This study aims to address the effect of JP in MRL/lpr mice and elucidate the pharmacological mechanism by which JP targets CD11a and CD70 DNA methylation via the miR-29b-sp1/DNMT1 pathway.. Mice treated with JP exhibited a significant decrease in anti-dsDNA, TNF-α, TGF-β, and UP, accompanied by a significant increase in IL-2. HE staining revealed JP effectively mitigated renal inflammatory response, while Masson staining indicated a reduction in collagen fiber content. In addition, JP exhibited a significant impact on the global hypomethylation of SLE, as evidenced by the induction of high methylation levels of CD11a and CD70 promoter regions, mediated through the miR-29b-sp1/DNMT1 pathway.. Our findings demonstrate JP exerts a protective effect against spontaneous SLE development, attenuates renal pathological changes, and functions as a miRNA inhibitor to enhance CD11a and CD70 DNA methylation through the modulation of the miR-29b-sp1/DNMT1 pathway.

Topics: Animals; CD4-Positive T-Lymphocytes; DNA Methylation; Interleukin-2; Lupus Erythematosus, Systemic; Mice; Mice, Inbred C57BL; Mice, Inbred MRL lpr; MicroRNAs; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Our group has previously demonstrated elevated serum-soluble ST2 in patients with active systemic lupus erythematosus, suggesting a role of IL-33 in the underlying pathogenesis. However, inconsistent results have been reported on the effect of exogenous IL-33 on murine lupus activity, which may be mediated by concerted actions of various immune cells in vivo. This study aimed to examine the function of IL-33 on macrophage polarization and regulatory T cells (Treg) and their interactive effects in the lupus setting by in vitro coculture experiments of macrophages and T cells that were performed in the presence or absence of IL-33-containing medium. Compared to IL-4-polarized bone marrow-derived macrophages (BMDM) from MRL/MpJ mice, adding IL-33 enhanced mRNA expression of markers of alternatively activated macrophages, including CD206 and Arg1. IL-33 and IL-4 copolarized BMDM produced higher TGF-β but not IL-6 upon inflammatory challenge. These BMDM induced an increase in the Foxp3+CD25+ Treg population in cocultured allogeneic T cells from MRL/MpJ and predisease MRL/lpr mice. These copolarized BMDM also showed an enhanced suppressive effect on T cell proliferation with reduced IFN-γ and IL-17 release but increased TGF-β production. In the presence of TGF-β and IL-2, IL-33 also directly promoted inducible Treg that expressed a high level of CD25 and more sustained Foxp3. Unpolarized BMDM cocultured with these Treg displayed higher phagocytosis. In conclusion, TGF-β was identified as a key cytokine produced by IL-4 and IL-33 copolarized alternatively activated macrophages and the induced Treg, which may contribute to a positive feedback loop potentiating the immunoregulatory functions of IL-33.

Topics: Animals; Forkhead Transcription Factors; Interleukin-33; Interleukin-4; Lupus Erythematosus, Systemic; Macrophages; Mice; Mice, Inbred MRL lpr; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is propelled by pathogenic autoantibody (AutoAb) and immune pathway dysregulation. Identifying populations at risk of reaching classified SLE is essential to curtail inflammatory damage. Lupus blood relatives (Rel) have an increased risk of developing SLE. We tested factors to identify Rel at risk of developing incomplete lupus (ILE) or classified SLE vs. clinically unaffected Rel and healthy controls (HC), drawing from two unique, well characterized lupus cohorts, the lupus autoimmunity in relatives (LAUREL) follow-up cohort, consisting of Rel meeting <4 ACR criteria at baseline, and the Lupus Family Registry and Repository (LFRR), made up of SLE patients, lupus Rel, and HC. Medical record review determined ACR SLE classification criteria; study participants completed the SLE portion of the connective tissue disease questionnaire (SLE-CSQ), type 2 symptom questions, and provided samples for assessment of serum SLE-associated AutoAb specificities and 52 plasma immune mediators. Elevated SLE-CSQ scores were associated with type 2 symptoms, ACR scores, and serology in both cohorts. Fatigue at BL was associated with transition to classified SLE in the LAUREL cohort (

Topics: Autoantibodies; Autoimmunity; Humans; Inflammation; Interleukin-10; Lupus Erythematosus, Systemic; Self Report; Surveys and Questionnaires; Transforming Growth Factor beta

To investigate the differentiation of regulatory T cells (Tregs) induced by methylprednisolone (MP) pulse therapy in patients with Systemic Lupus Erythematosus (SLE).. We enrolled 30 patients with SLE and analyzed peripheral blood mononuclear cells (PBMCs) before and after MP pulse therapy. Peripheral Tregs, apoptosis of PBMCs subsets, and TGFβ production by monocytes was quantified by flow cytometry. Proliferation and IFN-γ production of CD4. Peripheral Tregs was significantly increased after MP pulse therapy (6.76 ± 1.46% vs. 3.82 ± 1.02%, p < 0.01), with an expansion of Nrp1. MP pulse therapy induces CD4

Topics: Apoptosis; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Methylprednisolone; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

An interplay between immune cells and resident skin and joint stromal cells is implicated in psoriatic arthritis (PsA), yet the mechanisms remain elusive with a paucity of molecular biomarkers for activity and response. Combined transcriptomic and immunophenotypic analysis of whole blood and skin fibroblasts could provide further insights.. Whole blood RNA-seq was performed longitudinally in 30 subjects with PsA at the beginning, one and six months after treatment, with response defined at six months. As control groups, 10 healthy individuals and 10 subjects with rheumatoid arthritis (RA) were recruited combined with public datasets from patients with psoriasis (PsO) and systemic lupus erythematous (SLE). Differential expression analysis and weighted gene co-expression network analysis were performed to identify gene expression signatures, while deconvolution and flow cytometry to characterize the peripheral blood immune cell profile. In a subset of affected and healthy individuals, RNA-seq of skin fibroblasts was performed and subjected to CellChat analysis to identify the blood-skin fibroblast interaction network.. PsA demonstrated a distinct "activity" gene signature in the peripheral blood dominated by TNF- and IFN-driven inflammation, deregulated cholesterol and fatty acid metabolism and expansion of pro-inflammatory non-classical monocytes. Comparison with the blood transcriptome of RA, PsO, and SLE revealed a ". Transcriptome analysis of peripheral blood and skin fibroblasts in PsA reveals a distinct disease activity signature and supports the involvement of skin fibroblasts through their activation and interaction with circulating immune cells. Aberrant TGFβ signaling and persistently increased non-classical monocytes characterize treatment-resistant PsA, with pro-inflammatory pathways related to platelet activation and Hippo signaling predicting early response to treatment.

Topics: Arthritis, Psoriatic; Arthritis, Rheumatoid; Biomarkers; Fatty Acids; Fibroblasts; Gene Expression Profiling; Humans; Lupus Erythematosus, Systemic; Psoriasis; Semaphorins; Transcriptome; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a chronic autoimmune inflammatory disease characterized by the involvement of multiple organs. Lupus nephritis (LN) is a major risk factor for overall morbidity and mortality in SLE patients. Hence, designing effective drugs is pivotal for treating individuals with LN. Fisetin plays a senolytic role by specifically eliminating senescent cells, inhibiting cell proliferation, and exerting anti-inflammatory, anti-oxidant, and anti-tumorigenic effects. However, limited research has been conducted on the utility and therapeutic mechanisms of fisetin in chronic inflammation. Similarly, whether the effects of fisetin depend on cell type remains unclear. In this study, we found that LN-prone MRL/lpr mice demonstrated accumulation of Ki-67-positive myofibroblasts and p15

Topics: Animals; Antioxidants; Epithelial Cells; Fibroblasts; Lupus Erythematosus, Systemic; Lupus Nephritis; Mice; Mice, Inbred MRL lpr; Transforming Growth Factor beta

The Treg/Th17 imbalance is associated with the development of systemic lupus erythematosus (SLE). Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin, is isolated from the traditional Chinese herb Artemisia annua Artemisia annua L. This study aims to evaluate the effects of DHA alone or in combination with prednisone in immunodeficiency of SLE. In vivo, the therapeutical effect of DHA alone or in combination with prednisone was assessed in the pristane-induced SLE mouse model. Then, the level of serum antibodies, creatinine (Cre), blood urea nitrogen (BUN), urine protein, kidney histopathology, interleukin (IL)-17, IL-6, transforming growth factor (TGF)-β, the expression of RORγt and Foxp3, the percentages of Treg and Th17 in peripheral blood and spleen were assayed. In vitro, the mouse spleen lymphocytes were separated and treated with DHA alone or DHA in combination with prednisone. Then the percentages of Treg and Th17, the concentration of IL-17, IL-6, TGF-β, and the expression of RORγt and Foxp3 were assayed. It was shown that DHA alone or in combination with prednisone treatment significantly alleviated the manifestations of pristane-induced SLE mice, suppressed inflammation and restored the Treg/Th17 balance. DHA alone or in combination with prednisone significantly inhibited Th17 cell differentiation while induced Treg cell differentiation in vitro. DHA alone or in combination with prednisone also reduced the transcription of RORγt and increased Foxp3 in lymphocytes, as well as IL-17 and TGF-β levels. Our data indicated that DHA can produce synergistic effect with prednisone to attenuate the symptoms of SLE by restoring Treg/Th17 balance.

Topics: Animals; Artemisinins; Cell Differentiation; Lupus Erythematosus, Systemic; Mice; Prednisone; Th17 Cells; Transforming Growth Factor beta

Defects causing concomitant loss of CD25 expression in regulatory T cells (Tregs) have been identified in systemic lupus erythematosus (SLE). However, the cause of this deficiency is not fully understood. Carcinoembryonic antigen related cell adhesion molecule 1 (CEACAM1), an immune co-receptor, contributes to general T-cell function and activation. Our previous study revealed that CEACAM1 expression was upregulated in peripheral blood mononuclear cells (PBMCs) from patients with SLE. However, its role remains unclear. Herein, we confirmed CEACAM1, especially CEACAM1-S, was upregulated in PBMCs from patients with SLE. CEACAM1-S over-expression inhibits CD4

Topics: Antigens, CD; Cell Adhesion Molecule-1; Cell Adhesion Molecules; Cell Differentiation; Flow Cytometry; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; MicroRNAs; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Immune cells play important roles in systemic lupus erythematosus (SLE). We previously found that myeloid-derived suppressor cell (MDSC)-derived arginase-1 (Arg-1) promoted Th17 cell differentiation in SLE. In the present study, we performed RNA-chip to identify the microRNA regulation network between MDSCs and Th17 cells. miR-542-5p in humans, as the homologous gene of miR-322-5p in mice was significantly up-regulated in the Th17+MDSC group compared with Th17 cells cultured alone and down-regulated in the Th17+MDSC+Arg-1 inhibitor group compared with the Th17+MDSC group. We further evaluated the miR-322-5p and Th17/Treg balance in mice and found that the proportions of both Th17 cells and Tregs were elevated and that miR-322-5p overexpression activated the transforming growth factor-β pathway. Moreover, although miR-322-5p expression was higher in SLE mice, it decreased after treatment with an Arg-1 inhibitor. The proportion of Th17 cells and Th17/Treg ratio correlated with miR-322-5p levels. In conclusion, MDSC-derived Arg-1 and mmu-miR-322-5p not only promote Th17 cell and Treg differentiation, but also shift the Th17/Treg ratio in SLE. The Arg-1/miR-322-5p axis may serve as a novel treatment target for SLE.

Topics: Animals; Arginase; Disease Models, Animal; Disease Progression; Female; Gene Expression; Humans; Lupus Erythematosus, Systemic; Mice, Inbred C57BL; MicroRNAs; Myeloid-Derived Suppressor Cells; Signal Transduction; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

To develop a nanoparticle (NP) platform that can expand both CD4+ and CD8+ Treg cells in vivo for the suppression of autoimmune responses in systemic lupus erythematosus (SLE).. Poly(lactic-co-glycolic acid) (PLGA) NPs encapsulating interleukin-2 (IL-2) and transforming growth factor β (TGFβ) were coated with anti-CD2/CD4 antibodies and administered to mice with lupus-like disease induced by the transfer of DBA/2 T cells into (C57BL/6 × DBA/2)F. Anti-CD2/CD4 antibody-coated, but not noncoated, NPs encapsulating IL-2 and TGFβ induced CD4+ and CD8+ FoxP3+ Treg cells in vitro. The optimal dosing regimen of NPs for expansion of CD4+ and CD8+ Treg cells was determined in in vivo studies in mice without lupus and then tested in BDF1 mice with lupus. The administration of anti-CD2/CD4 antibody-coated NPs encapsulating IL-2 and TGFβ resulted in the expansion of CD4+ and CD8+ Treg cells, a marked suppression of anti-DNA antibody production, and reduced renal disease.. This study shows for the first time that T cell-targeted PLGA NPs encapsulating IL-2 and TGFβ can expand both CD4+ and CD8+ Treg cells in vivo and suppress murine lupus. This approach, which enables the expansion of Treg cells in vivo and inhibits pathogenic immune responses in SLE, could represent a potential new therapeutic modality in autoimmune conditions characterized by impaired Treg cell function associated with IL-2 deficiency.

Topics: Animals; CD8-Positive T-Lymphocytes; Disease Models, Animal; Dose-Response Relationship, Drug; Interleukin-2; Lupus Erythematosus, Systemic; Mice; Mice, Inbred C57BL; Nanoparticles; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

To identify single-cell transcriptional signatures of dendritic cells (DCs) that are associated with autoimmunity, and determine whether those DC signatures are correlated with the clinical heterogeneity of autoimmune disease.. Blood-derived DCs were single-cell sorted from the peripheral blood of patients with rheumatoid arthritis, systemic lupus erythematosus, or type 1 diabetes as well as healthy individuals. DCs were analyzed using single-cell gene expression assays, performed immediately after isolation or after in vitro stimulation of the cells. In addition, protein expression was measured using fluorescence-activated cell sorting.. CD1c+ conventional DCs and plasmacytoid DCs from healthy individuals exhibited diverse transcriptional signatures, while the DC transcriptional signatures in patients with autoimmune disease were altered. In particular, distinct DC clusters, characterized by up-regulation of TAP1, IRF7, and IFNAR1, were abundant in patients with systemic autoimmune disease, whereas DCs from patients with type 1 diabetes had decreased expression of the regulatory genes PTPN6, TGFB, and TYROBP. The frequency of CD1c+ conventional DCs that expressed a systemic autoimmune profile directly correlated with the extent of disease activity in patients with rheumatoid arthritis (Spearman's r = 0.60, P = 0.03).. DC transcriptional signatures are altered in patients with autoimmune disease and are associated with the level of disease activity, suggesting that immune cell transcriptional profiling could improve our ability to detect and understand the heterogeneity of these diseases, and could guide treatment choices in patients with a complex autoimmune disease.

Topics: Adaptor Proteins, Signal Transducing; Arthritis, Rheumatoid; ATP Binding Cassette Transporter, Subfamily B, Member 2; Autoimmune Diseases; Case-Control Studies; Dendritic Cells; Diabetes Mellitus, Type 1; Flow Cytometry; Gene Expression Profiling; Humans; Inflammation; Interferon Regulatory Factor-7; Lupus Erythematosus, Systemic; Membrane Proteins; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Receptor, Interferon alpha-beta; Severity of Illness Index; Single-Cell Analysis; Transforming Growth Factor beta; Up-Regulation

Allergic and autoimmune diseases comprise a group of inflammatory disorders caused by aberrant immune responses in which CD25+ Forkhead box P3-positive (FOXP3+) T regulatory (Treg) cells that normally suppress inflammatory events are often poorly functioning. This has stimulated an intensive investigative effort to find ways of increasing Tregs as a method of therapy for these conditions. One such line of investigation includes the study of how ligation of Toll-like receptors (TLRs) by CpG oligonucleotides (ODN) results in an immunostimulatory cascade that leads to induction of T-helper (Th) type 1 and Treg-type immune responses.. The present study investigated the mechanisms by which calf thymus mammalian double-stranded DNA (CT-DNA) and a synthetic methylated DNA CpG ODN sequence suppress in vitro lymphoproliferative responses to antigens, mitogens, and alloantigens when measured by [3H]-thymidine incorporation and promote FoxP3 expression in human CD4+ T cells in the presence of transforming growth factor (TGF) beta and interleukin-2 (IL-2).. Lymphoproliferative responses of peripheral blood mononuclear cells from four healthy subjects or nine subjects with systemic lupus erythematosus to CT-DNA or phytohemagglutinin (PHA) was measured by tritiated thymidine ([3H]-TdR) incorporation expressed as a stimulation index. Mechanisms of immunosuppressive effects of CT-DNA were evaluated by measurement of the degree of inhibition to lymphoproliferative responses to streptokinase-streptodornase, phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), or alloantigens by a Con A suppressor assay. The effects of CpG methylation on induction of FoxP3 expression in human T cells were measured by comparing inhibitory responses of synthetic methylated and nonmethylated 8-mer CpG ODN sequences by using cell sorting, in vitro stimulation, and suppressor assay.. Here, we showed that CT-DNA and a synthetic methylated DNA 8-mer sequence could suppress antigen-, mitogen-, and alloantigen-induced lymphoproliferation in vitro when measured by [3H]-thymidine. The synthetic methylated DNA CpG ODN but not an unmethylated CpG ODN sequence was shown to promote FoxP3 expression in human CD4+ T cells in the presence of TGF beta and IL-2. The induction of FoxP3+ suppressor cells is dose dependent and offers a potential clinical therapeutic application in allergic and autoimmune and inflammatory diseases.. The use of this methylated CpG ODN offers a broad clinical application as a novel therapeutic method for Treg induction and, because of its low cost and small size, should facilitate delivery via nasal, respiratory, gastrointestinal routes, and/or by injection, routes of administration important for vaccine delivery to target sites responsible for respiratory, gastrointestinal, and systemic forms of allergic and autoimmune disease.

Topics: Animals; Cattle; CD4-Positive T-Lymphocytes; Cell Proliferation; Cells, Cultured; CpG Islands; DNA; DNA Methylation; Forkhead Transcription Factors; Humans; Hypersensitivity; Immunosuppression Therapy; Immunotherapy; Isoantigens; Lupus Erythematosus, Systemic; Lymphocyte Activation; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

The molecular targets of the vast majority of autoantibodies in systemic lupus erythematosus (SLE) are unknown. We set out to identify novel autoantibodies in SLE to improve diagnosis and identify subgroups of SLE individuals.. A baculovirus-insect cell expression system was used to create an advanced protein microarray with 1543 full-length human proteins expressed with a biotin carboxyl carrier protein (BCCP) folding tag, to enrich for correctly folded proteins. Sera from a discovery cohort of UK and US SLE individuals (n = 186) and age/ethnicity matched controls (n = 188) were assayed using the microarray to identify novel autoantibodies. Autoantibodies were validated in a second validation cohort (91 SLE, 92 controls) and a confounding rheumatic disease cohort (n = 92).. We confirmed 68 novel proteins as autoantigens in SLE and 11 previous autoantigens in both cohorts (FDR<0.05). Using hierarchical clustering and principal component analysis, we observed four subgroups of SLE individuals associated with four corresponding clusters of functionally linked autoantigens. Two clusters of novel autoantigens revealed distinctive networks of interacting proteins: SMAD2, SMAD5 and proteins linked to TGF-β signalling; and MyD88 and proteins involved in TLR signalling, apoptosis, NF-κB regulation and lymphocyte development. The autoantibody clusters were associated with different patterns of organ involvement (arthritis, pulmonary, renal and neurological). A panel of 26 autoantibodies, which accounted for four SLE clusters, showed improved diagnostic accuracy compared to conventional antinuclear antibody and anti-dsDNA antibody assays.. These data suggest that the novel SLE autoantibody clusters may be of prognostic utility for predicting organ involvement in SLE patients and for stratifying SLE patients for specific therapies.

Topics: Adult; Animals; Antibodies, Antinuclear; Autoantigens; Baculoviridae; Cohort Studies; Female; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; NF-kappa B; Prognosis; Protein Array Analysis; Protein Interaction Maps; Sf9 Cells; Signal Transduction; Smad2 Protein; Smad5 Protein; Toll-Like Receptors; Transforming Growth Factor beta

We aimed at investigating whether the frequency and function of T helper 17 (Th17) and regulatory T cells (Treg) are affected by a restriction of dietary sodium intake in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). We enrolled RA and SLE patients not receiving drugs known to increase urinary sodium excretion. Patients underwent a dietary regimen starting with a restricted daily sodium intake followed by a normal-sodium daily intake. The timepoints were identified at baseline (T0), after 3 weeks of low-sodium dietary regimen (T3), after 2 weeks of normal-sodium dietary regimen (T5). On these visits, we measured the 24-hour urinary sodium excretion, the frequency and function of Th17 and Treg cells in the peripheral blood, the serum levels of cytokines. Analysis of urinary sodium excretion confirmed adherence to the dietary regimen. In RA patients, a trend toward a reduction in the frequencies of Th17 cells over the low-sodium dietary regimen followed by an increase at T5 was observed, while Treg cells exhibited the opposite trend. SLE patients showed a progressive reduction in the percentage of Th17 cells that reached a significance at T5 compared to T0 (p = 0.01) and an increase in the percentage of Treg cells following the low-sodium dietary regimen at both T1 and T3 compared to T0 (p = 0.04 and p = 0.02, respectively). No significant apoptosis or proliferation modulation was found. In RA patients, we found a reduction at T5 compared to T0 in serum levels of both TGFβ (p = 0.0016) and IL-9 (p = 0.0007); serum IL-9 levels were also reduced in SLE patients at T5 with respect to T0 (p = 0.03). This is the first study investigating the effects of dietary sodium intake on adaptive immunity. Based on the results, we hypothesize that a restricted sodium dietary intake may dampen the inflammatory response in RA and SLE patients.

Topics: Aged; Apoptosis; Arthritis, Rheumatoid; Cell Proliferation; Cytokines; Diet; Enzyme-Linked Immunosorbent Assay; Female; Humans; Inflammation; Interleukin-9; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Male; Middle Aged; Sodium, Dietary; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

The present study aimed to examine the status and clinical significance of cluster of differentiation (CD) 19+CD24highCD38high regulatory B cells (Bregs), serum interleukin (IL)‑10, serum transforming growth factor (TGF)‑β and IL‑10 receptor (IL‑10R) expression in peripheral blood from patients with systemic lupus erythematosus (SLE). A total of 56 patients with SLE and 35 healthy individuals were recruited to the present study. The SLE disease activity index (SLEDAI) was calculated, and other laboratory parameters were measured. Peripheral blood was collected from all participants. The frequency of CD19+CD24highCD38high Bregs and IL‑10R+ expression on circulating lymphocytes was examined by flow cytometry. The serum levels of IL‑10 and TGF‑β were measured using enzyme‑linked immunosorbent assay. The associations between these measurements and SLEDAI or other laboratory parameters were analyzed by correlation analysis. The percentage of CD19+CD24highCD38high Bregs and the serum levels of IL‑10 were significantly increased, whereas the expression of IL‑10R on circulating lymphocytes was markedly reduced in patients with SLE compared with in healthy controls. The serum levels of TGF‑β1 were not markedly different between the groups. In addition, these factors were correlated with other SLE laboratory parameters, and inter‑correlations were presented with different degrees of significance. The percentage of CD19+CD24highCD38high Bregs was positively correlated with the percentage of IL‑10R+ lymphocytes, mean fluorescence intensity (MFI) of IL‑10R+ lymphocytes and serum IL‑10 levels. In addition, the percentage of IL‑10R+ lymphocytes was positively correlated with its expression level (MFI), whereas serum TGF‑β1 levels were negatively correlated with serum IL‑10 levels. The present results indicated that expansion of CD19+CD24highCD38high Bregs, upregulation of IL‑10 and deficient lymphocyte‑associated IL‑10R may serve as novel SLE biomarkers. It may be hypothesized that deficient IL‑10R expression results in compensatory enhanced IL‑10 expression, expansion of Bregs, and/or compromised Breg and IL‑10 functions, thus contributing to SLE development. Therefore, targeting the 'Bregs/IL‑10/IL‑10R' system may provide a novel therapeutic approach for the treatment of SLE.

Topics: Adult; Antigens, CD; B-Lymphocytes; Biomarkers; Extracellular Matrix Proteins; Female; Humans; Interleukin-10; Lupus Erythematosus, Systemic; Male; Transforming Growth Factor beta; Up-Regulation

Imbalanced cellular immunity is critical to the pathogenesis of systemic lupus erythematosus (SLE). Recently, autophagy has emerged as a key homeostatic mechanism in T lymphocytes. This study was conducted to explore the impact of autophagy on the Th17/ regulatory T (Treg) immune imbalance in SLE.. Peripheral Th17 and Treg cells from newly diagnosed patients with SLE and healthy controls were detected by flow cytometry. Additionally, the effects of chloroquine (CQ) autophagic inhibition on the Th17/Treg immune response were investigated in vitro. In addition, hydroxychloroquine (HCQ) treatment of the Th17/Treg immune response and the disease progression of lupus MRL/lpr mice were studied in vivo.. Compared with healthy controls, both peripheral Th17 and Treg cells of patients with SLE exhibited activated autophagy, resulting in a heightened Th17 proinflammatory response and diminished Treg immunosuppression. Furthermore, in vitro experiments indicated that CQ autophagic inhibition effectively rebalanced the Th17/Treg immune responses in patients with SLE. In vivo studies of MRL/lpr mice similarly confirmed that HCQ treatment decisively inhibited the autophagy of Th17/Treg cellular subsets, restoring the immune balance, lowering the serum levels of inflammatory cytokines and autoantibodies, and improving renal histopathology.. Activated autophagy contributed to the Th17/Treg immune imbalance in SLE, and chloroquine autophagic inhibition rebalanced Th17/ Treg-mediated immunity and ameliorated SLE.

Topics: Adult; Animals; Antimalarials; Autophagy; Cells, Cultured; Chloroquine; Female; Humans; Hydroxychloroquine; Interferon-gamma; Interleukin-17; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Male; Mice; Mice, Inbred MRL lpr; Microtubule-Associated Proteins; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta; Young Adult

Efficient clearance of apoptotic cells (AC) is pivotal in preventing autoimmunity and is a potent immunosuppressive stimulus. However, activation of cells prior to apoptosis abolishes their immunoregulatory properties. Here we show using the antigen-induced model of arthritis that the degree of DNA methylation within AC confers their immunomodulatory plasticity. DNA isolated from resting and activated AC mimicked their respective immune effects. Demethylation of DNA abrogated the protective effect of AC whereas remethylation of AC DNA reversed the effects of activation and restored the ability to inhibit inflammation. Disease suppression or lack thereof was associated with TGFβ and IL-6 production respectively. Apoptotic CD4

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; CD4-Positive T-Lymphocytes; DNA; DNA Methylation; Epigenesis, Genetic; Gene Expression Regulation; Humans; Interleukin-6; Lipopolysaccharides; Lupus Erythematosus, Systemic; Macrophages; Mice; Mice, Inbred C57BL; Phagocytosis; Primary Cell Culture; Serum Albumin, Bovine; Signal Transduction; Transforming Growth Factor beta

Long non-coding RNAs (lncRNAs) are thought to have various functions other than RNA silencing. We tried to evaluate the expression of lncRNAs in patients with systemic sclerosis (SSc) and determined whether lncRNAs controls collagen expression in dermal fibroblasts. lncRNA expression was determined by real-time PCR and in situ hybridization. Protein and mRNA levels of collagen were analysed using immunoblotting and real-time PCR. We found TSIX, one of the lncRNAs, was overexpressed in SSc dermal fibroblasts both in vivo and in vitro, which was inhibited by the transfection of transforming growth factor (TGF)-β1 siRNA. TSIX siRNA reduced the mRNA expression of type I collagen in normal and SSc dermal fibroblasts, but not the levels of major disease-related cytokines. In addition, TSIX siRNA significantly reduced type I collagen mRNA stability, but not protein half-lives. Furthermore, we first investigated serum lncRNA levels in patients with SSc, and serum TSIX levels were significantly increased in SSc patients. TSIX is a new regulator of collagen expression which stabilizes the collagen mRNA. The upregulation of TSIX seen in SSc fibroblasts may result from activated endogenous TGF-β signalling and may play a role in the constitutive upregulation of collagen in these cells. Further studies on the regulatory mechanism of tissue fibrosis by lncRNAs in SSc skin lead to a better understanding of the pathogenesis, new diagnostic methods by their serum levels and new therapeutic approaches using siRNAs.

Topics: Adult; Aged; Aged, 80 and over; Collagen Type I; Dermis; Female; Fibroblasts; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; RNA Interference; RNA Stability; RNA, Long Noncoding; RNA, Messenger; RNA, Small Interfering; Scleroderma, Systemic; Signal Transduction; Transforming Growth Factor beta; Up-Regulation; Young Adult

Risk posed by microbial superantigens in triggering or exacerbating SLE in genetically predisposed individuals, thereby altering the response to its treatment strategies, has not been studied. Using streptococcal pyrogenic exotoxin A and staphylococcal enterotoxin B as prototype superantigens, we have demonstrated that they profoundly affect the magnitude of polyclonal T cell response, particularly CD4(+) T cells and expression of CD45RA and CD45RO, and cytokine secretion in vitro in SLE patient PBMCs. Also, reduced proportions of FoxP3 expressing CD4(+)CD25(+) T cells were detected in SLE as compared to healthy control PBMCs. Furthermore, polymorphism in IL-10 and TGF-β showed significant association with SLE in our study population. These results indicate that accumulation of superantigen-reactive T cells and cytokine polymorphism may cause disease exacerbation, relapse, or therapeutic resistance in SLE patients. Attempts to contain colonizing and/or superantigen-producing microbial agents in SLE patients in addition to careful monitoring of their therapy may be worthwhile in decreasing disease severity or preventing frequent relapses. The study suggests that superantigen interference in conjunction with cytokine polymorphism may play a role in immune dysregulation, thereby contributing to autoimmunity in SLE. Therefore, changes in T cell phenotypes and cytokine secretion might be good indicators of therapeutic efficacy in these patients.

Topics: Adult; Antibodies, Antinuclear; Bacterial Proteins; Bacterial Toxins; CD4-Positive T-Lymphocytes; Cells, Cultured; Disease Progression; Exotoxins; Female; Forkhead Transcription Factors; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-10; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Membrane Proteins; Polymorphism, Genetic; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Young Adult

Foxp3(+) regulatory T cells (Treg) playing a crucial role in the maintenance of immune tolerance and prevention of autoimmune diseases consist of thymus-derived naturally occurring CD4(+)Foxp3(+) Treg cells (nTreg) and those that can be induced ex vivo with TGF-β (iTreg). Although both Treg subsets share similar phenotypes and functional characteristics, they also have potential biologic differences on their biology. The role of iTreg in regulating B cells remains unclear so far. The suppression assays of Treg subsets on activation, proliferation, and Abs production of B cells were measured using a Treg and B cell coculture system in vitro. Transwell and Ab blockade experiments were performed to assess the roles of cell contact and soluble cytokines. Treg were adoptively transferred to lupus mice to assess in vivo effects on B cells. Like nTreg, iTreg subset also directly suppressed activation and proliferation of B cells. nTreg subset suppressed B cell responses through cytotoxic manner related to expression of granzyme A, granzyme B, and perforin, whereas the role of iTreg subset on B cells did not involve in cytotoxic action but depending on TGF-β signaling. Furthermore, iTreg subset can significantly suppress Ab produced by lupus B cells in vitro. Comparison experiments using autoantibodies microarrays demonstrated that adoptive transfer of iTreg had a superior effect than nTreg subset on suppressing lupus B cell responses in vivo. Our data implicate a role and advantage of iTreg subset in treating B cell-mediated autoimmune diseases, boosting the translational potential of these findings.

Topics: Adoptive Transfer; Animals; Autoantibodies; B-Lymphocytes; Cell Differentiation; Cell Proliferation; Coculture Techniques; Granzymes; Immune Tolerance; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Oligonucleotide Array Sequence Analysis; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease which is characterized by the presence of auto-reactive T cell and anti-ds DNA antibodies. Treg cells are crucial for maintaining immunologic self-tolerance and are shown to be reduced in SLE patients. 1,25-Dihydroxyvitamin D3 has immunomedulatory effects on the immune system and has recently received substantial attention.. In this study we evaluated the effects of 1,25-dihydroxyvitamin D3 on Treg cells and related cytokines in lupus-like induced mice model.. Female Balb/c mice were divided into four groups: Group one: injected with PBS and Freund's adjuvant; Group two: injected with non-activated chromatin; Group three: Lupus-like disease was induced with activated chromatin; Group four: Mice were initially treated for two weeks with 1,25-dihydroxyvitamin D3 and then lupus-like disease was induced. Group five: Four mice from group one were treated with 1,25-dihydroxyvitamin D3 for two weeks after disease establishment. Ten weeks after the last injection the mice were killed and spleens were studied for Treg percentages and expression of cytokine genes.. We found that treatment with 1,25-dihydroxyvitamin D3 reduces IL-6 and IL-10 mRNA expression and increases TGF-β and Foxp3 mRNA expression levels, and also enhances spleen Treg percentage.. The remarkable reduction of IL-6 and IL-10 gene expressions, significant enhancement of TGF-β and Foxp3 gene expressions, along with an increase in Treg cell population after oral 1,25-dihydroxyvitamin D3 administration suggest a possible role for this vitamin as a prophylactic supplement in SLE.

Topics: Animals; Antibodies, Antinuclear; Calcitriol; Disease Models, Animal; DNA; Female; Forkhead Transcription Factors; Gene Expression; Immunologic Factors; Interleukin-10; Interleukin-6; Lupus Erythematosus, Systemic; Lymphocyte Count; Mice, Inbred BALB C; Spleen; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

We aimed to elucidate the roles of six pro-angiogenic factors, namely, basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), placental growth factor (PlGF), hepatocyte growth factor (HGF), transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha), in the pathogenesis of systemic lupus erythematosus (SLE).. A group of 75 patients with SLE and a control group of 40 healthy volunteers were recruited. Angiogenic factors were tested through enzyme-linked immunosorbent assay (ELISA) measurement. The angiogenic activities of the patients with SLE and the healthy controls were assessed and a correlation analysis of these angiogenic factors was conducted.. A much higher level of angiogenic activity was shown in the serum of patients with SLE than that of the healthy controls, yet no statistically significant difference was found in the angiogenic activities of active SLE and inactive SLE. Serum VEGF level in the active SLE group was significantly higher than that in the control group and the inactive SLE group, and serum HGF level was strongly positively correlated with VEGF in all SLE groups; meanwhile, there was also a statistically significant positive correlation between TNF-alpha and VEGF in all SLE groups. There was a statistically significant positive correlation between serum VEGF level and bFGF level in the active SLE group. There was a slightly negative correlation between serum HGF level and TGF-beta level in the SLE group, but this negativity did not reach the significance level. Likewise, positive correlation was also shown in the serum VEGF level and PlGF level, yet not in bFGF with PlGF.. Circulating serum angiogenic cytokines may be disease markers of SLE activity. Anti-angiogenic agents such as thalidomide and endogenous angiogenesis inhibitors such as endostatin are potentially effective and promising therapies in the treatment of SLE.

Topics: Adult; Angiogenesis Inhibitors; Autoantibodies; Biomarkers; Endostatins; Female; Fibroblast Growth Factor 2; Hepatocyte Growth Factor; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Neovascularization, Pathologic; Placenta Growth Factor; Pregnancy Proteins; Thalidomide; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

As abundant types of genetic predisposition and environmental factors seem to be associated with the development of juvenile-onset systemic lupus erythematosus (JSLE), we investigated the gene polymorphisms of two anti-inflammatory cytokines, including interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β), which were previously found to be associated with SLE in adults, in a group of patients with JSLE. We studied a group of 59 Iranian patients with JSLE in comparison with 140 healthy controls and assessed the frequency of alleles, genotypes, and haplotypes of IL-10 and TGF-β single-nucleotide polymorphisms (SNPs) using polymerase chain reaction with sequence-specific primers method. The CA genotype was significantly more frequent at position -592 in IL-10 in patients with juvenile-onset systemic lupus erythematosus than in the controls (P = 0.01). Genotype CC was detected at the same position in 32.7 % of the patients; this frequency was significantly lower than the frequency of 50.7 % recorded in the healthy controls (P = 0.03). The TC haplotype of TGF-β (codon 10, codon 25) was significantly more frequent in the patients with juvenile-onset systemic lupus erythematosus than in the healthy controls (P = 0.02). Nevertheless, these significant associations disappear after Bonferroni correction. Our findings suggest that IL-10 (-1082, -819, -592) and TGF-β (codon 10, codon 25) gene variants may not be associated with the development of JSLE in Iranian population.

Topics: Adolescent; Age of Onset; Case-Control Studies; Child; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Humans; Interleukin-10; Iran; Lupus Erythematosus, Systemic; Male; Polymorphism, Single Nucleotide; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease. Emerging data suggests that T helper 17 (Th17) cells play a pathogenic role in SLE and the increased number of these cells correlates with disease activity. In recent years, 1α, 25-dihydroxyvitamin D3 (1,25VitD3) has been considered as an immunomodulatory factor.. To investigate the effect of 1,25VitD3 on Th17 cells and on the expression of related cytokines in SLE patients.. Thirty SLE patients (newly diagnosed or in remission) were sampled for 10 ml whole blood to isolate peripheral blood mononuclear cells (PBMCs) using Ficoll-Hypaque density gradient centrifugation. Isolated cells were cultured in the presence and absence of 50 nM 1,25VitD3. After incubation, cells were harvested and stimulated for 4-5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. IL-17 secreting cells were analyzed by flowcytometry. RNA was extracted from cultured cells, cDNA was synthesized, and the expression levels of IL-6, IL-17, IL-23 and TGF-β genes were assessed by real-time PCR.. The percentage of Th17 cells (CD3+CD8- IL-17+ T cells) decreased significantly in 1,25VitD3-treated cells (3.67 ± 2.43%) compared to untreated cells (4.65 ± 2.75%)( p=0.003). The expression of TGF-β up regulated (1.38-fold) and the expression of IL-6 (50%), IL-17 (27%) and IL-23 (64%) down regulated after 1,25VitD3 treatment.. This study showed that 1,25VitD3 modulates Th17 related pathways in SLE patients and revealed the immunomodulatory effect of 1,25VitD3 on the Th17 mediated autoimmunity.

Topics: Adult; Brefeldin A; Calcitriol; Cells, Cultured; Female; Flow Cytometry; Humans; Interleukin-17; Interleukin-23 Subunit p19; Interleukin-6; Ionomycin; Lupus Erythematosus, Systemic; Lymphocyte Count; Male; Tetradecanoylphorbol Acetate; Th17 Cells; Transforming Growth Factor beta

Programmed death-1 (PD-1) usually acts as a negative signal for T cell activation, and its expression on CD8(+)Foxp3(+) T cells is required for their suppressive capacity. In this study, we show that PD-1 signaling is required for the maintenance of functional regulatory CD4(+)CD25(+)Foxp3(+) regulatory T cells (CD4(+) T(reg)) that can control autoimmunity in (New Zealand Black × New Zealand White)F1 lupus mice. PD-1 signaling induced resistance to apoptosis and prolonged the survival of CD4(+) T(reg). In vivo, the blockade of PD-1 with a neutralizing Ab reduced PD-1 expression on CD4(+) T(reg) (PD1(lo)CD4(+) T(reg)). PD1(lo)CD4(+) T(reg) had an increased ability to promote B cell apoptosis and to suppress CD4(+) Th as compared with CD4(+) T(reg) with elevated PD-1 expression (PD1(hi)CD4(+) T(reg)). When PD-1 expression on CD4(+) T(reg) was blocked in vitro, PD1(lo)CD4(+) T(reg) suppressed B cell production of IgG and anti-dsDNA Ab. Finally, in vitro studies showed that the suppressive capacity of CD4(+) T(reg) depended on PD-1 expression and that a fine-tuning of the expression of this molecule directly affected cell survival and immune suppression. These results indicate that PD-1 expression has multiple effects on different immune cells that directly contribute to a modulation of autoimmune responses.

Topics: Adaptive Immunity; Animals; Antibodies, Monoclonal; Antibodies, Neutralizing; Autoantibodies; CD4 Antigens; Disease Progression; Female; Gene Expression Regulation; Immune Tolerance; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Programmed Cell Death 1 Receptor; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1-/Th2-type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2-type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL-4 (Th2), IFNγ (Th1), IL-10 and TGFβ by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30-expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFβ is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30-expressing T cells and IL-4, IFNγ, and immunosuppressive cytokines (IL-10 and TGFβ) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2-type response.

Topics: Adult; Biomarkers; CD3 Complex; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-8; Ki-1 Antigen; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; T-Lymphocyte Subsets; Transforming Growth Factor beta

Low-dose tolerance therapy with nucleosomal histone peptide epitopes blocks lupus disease in mouse models, but effect in humans is unknown. Herein, we found that CD4(+)CD25(high)FoxP3(+) or CD4(+)CD45RA(+)FoxP3(low) T-cells, and CD8(+)CD25(+)FoxP3(+) T-cells were all induced durably in PBMCs from inactive lupus patients and healthy subjects by the histone peptide/s themselves, but in active lupus, dexamethasone or hydroxychloroquine unmasked Treg-induction by the peptides. The peptide-induced Treg depended on TGFβ/ALK-5/pSmad 2/3 signaling, and they expressed TGF-β precursor LAP. Lupus patients' sera did not inhibit Treg induction. The peptide epitope-induced T cells markedly suppressed type I IFN related gene expression in lupus PBMC. Finally, the peptide epitopes suppressed pathogenic autoantibody production by PBMC from active lupus patients to baseline levels by additional mechanisms besides Treg induction, and as potently as anti-IL6 antibody. Thus, low-dose histone peptide epitopes block pathogenic autoimmune response in human lupus by multiple mechanisms to restore a stable immunoregulatory state.

Topics: Adult; Anti-Inflammatory Agents; Antigens, CD; Autoantibodies; Autoimmunity; CD8-Positive T-Lymphocytes; Epitopes; Female; Forkhead Transcription Factors; Gene Expression Regulation; Histones; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; Nucleosomes; Peptides; Primary Cell Culture; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad2 Protein; Smad3 Protein; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

To evaluate the specificity of expression patterns of cell-free circulating microRNAs (miRNAs) in systemic lupus erythematosus (SLE).. Total RNA was purified from plasma, and 45 different specific, mature miRNAs were determined using quantitative reverse transcription-polymerase chain reaction assays. A total of 409 plasma samples were obtained from 364 different patients with SLE, healthy control subjects, and control subjects with other autoimmune diseases. The results in the primary cohort of 62 patients with SLE and 29 healthy control subjects were validated in 2 independent cohorts: a validation cohort comprising 68 patients with SLE and 68 healthy control subjects, and a disease control cohort comprising 20 patients with SLE (19 of whom were from the other validation cohort), 46 healthy control subjects, 38 patients with vasculitis, 18 patients with rheumatoid arthritis, and 20 immunosuppressed patients.. Seven miRNAs were statistically significantly differentially expressed in plasma from patients with SLE. The expression of miRNA-142-3p (miR-142-3p) and miR-181a was increased, and the expression of miR-106a, miR-17, miR-20a, miR-203, and miR-92a was decreased. In addition, the expression of miR-342-3p, miR-223, and miR-20a was significantly decreased in SLE patients with active nephritis. A predictive model for SLE based on 2 or 4 miRNAs differentiated patients with SLE from control subjects (76% accuracy) when validated independently (P < 2 × 10(-9) ). Use of the 4-miRNA model provided highly significant differentiation between the SLE group and disease controls, except for those with vasculitis.. Circulating miRNAs are systematically altered in SLE. A 4-miRNA signature was diagnostic of SLE, and a specific subset of miRNA profiles was associated with nephritis. All of the signature miRNAs target genes in the transforming growth factor β signaling pathways. Other targets include regulation of apoptosis, cytokine-cytokine receptors, T cell development, and cytoskeletal organization. These findings highlight possible dysregulated pathways in SLE and suggest that circulating miRNA patterns distinguish SLE from other immunoinflammatory phenotypes.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Cohort Studies; Female; Gene Expression Profiling; Humans; Immunocompromised Host; Lupus Erythematosus, Systemic; Male; MicroRNAs; Middle Aged; Signal Transduction; Transforming Growth Factor beta; Vasculitis; Young Adult

Regulatory T (Treg) cells play an important role in the maintenance of immune tolerance to self and in the pathogenesis of autoimmune disease. Transforming growth factor-beta 1(TGF-β1) is a regulatory cytokine with pleiotropic properties in immune responses. This study was to investigate the role of Treg cells and TGF-β1 in the pathogenesis of patients with lupus nephritis (LN). A total of 42 new-onset systemic lupus erythematosus patients and 22 healthy controls were enrolled. The proportion of Treg cells in peripheral blood mononuclear cells (PBMCs) was evaluated by flow cytometric analysis. The serum and urinary TGF-β1 levels were measured by enzyme-linked immunosorbent assay (ELISA). The results demonstrated a significant decrease in the frequency of CD4(+)CD25(high) and CD4(+)CD25(+)FoxP3(+) T cells in LN patients. The concentration of serum TGF-β1 was found decreased in SLE patients, while urinary TGF-β1 levels were significantly higher in LN patients. Based on our results, decreased Treg cells were accompanied with lower serum TGF-β1 levels and higher urinary TGF-β1 levels in LN patients. TGF-ß1 levels in serum may play a key role in the pathogenesis of renal impairment while the significantly increased urinary TGF-β1 levels may be used as a biological marker in prediction of lupus nephritis.

Topics: Adult; Biomarkers; CD4 Antigens; Cell Count; Cell Separation; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Interleukin-2 Receptor alpha Subunit; Kidney; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; Middle Aged; Prognosis; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Monocytes in patients with systemic lupus erythematosus (SLE) are hyperstimulatory for T lymphocytes. We previously found that the normal program for expression of a negative costimulatory molecule programmed death ligand-1 (PD-L1) is defective in SLE patients with active disease. Here, we investigated the mechanism for PD-L1 dysregulation on lupus monocytes. We found that PD-L1 expression on cultured SLE monocytes correlated with TNF-α expression. Exogenous TNF-α restored PD-L1 expression on lupus monocytes. Conversely, TGF-β inversely correlated with PD-L1 in SLE and suppressed expression of PD-L1 on healthy monocytes. Therefore, PD-L1 expression in monocytes is regulated by opposing actions of TNF-α and TGF-β. As PD-L1 functions to fine tune lymphocyte activation, dysregulation of cytokines resulting in reduced expression could lead to loss of peripheral T cell tolerance.

Topics: B7-H1 Antigen; Humans; Lupus Erythematosus, Systemic; Monocytes; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

microRNAs (miRNAs) play a part in various cellular activities. However, the role of miRNA in SSc is not fully understood. This study investigated the expression and role of miR-92a in SSc patients and evaluated the possibility that miR-92a is involved in the pathogenesis of this disease.. Serum samples were obtained from 61 SSc patients. mRNAs were purified from serum and levels of miR-92a and miR-135 were measured with quantitative real-time PCR. miR-92a expression in dermal fibroblasts was also determined by quantitative real-time PCR. Immunoblotting was performed to detect MMP-1 protein.. The median serum levels of miR-92a, not miR-135, were significantly higher in SSc patients than normal subjects. The constitutive up-regulated miR-92a expression was also found in cultured dermal fibroblasts from SSc skin, which was decreased by the transfection with siRNA of TGF-β. Furthermore, the forced overexpression of miR-92a in normal dermal fibroblasts using miR-92a mimic resulted in the down-regulation of MMP-1 expression.. The increase of miR-92a in SSc may be due to the stimulation of intrinsic TGF-β activation seen in this disease. There is also a possibility that MMP-1 is the target of miR-92a and that increased miR-92a expression therefore plays a role in excessive collagen accumulation in SSc via the down-regulation of MMP-1. Clarifying the role of miRNAs in SSc may result in a better understanding of this disease and the development of new therapeutic approaches.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Cells, Cultured; Dermatomyositis; Dermis; Female; Fibroblasts; Gene Expression Regulation; Gene Silencing; Humans; Integrin alphaVbeta3; Lupus Erythematosus, Systemic; Male; Matrix Metalloproteinase 1; MicroRNAs; Middle Aged; RNA, Small Interfering; Scleroderma, Diffuse; Scleroderma, Localized; Transforming Growth Factor beta

Previous studies have indicated that bone marrow-derived mesenchymal stem cells (MSCs) from patients with systemic lupus erythematosus (SLE) exhibited impaired proliferation, differentiation, and immune modulation capacities. Thus, MSCs may be associated with the pathogenesis of SLE. The aim of this study was to determine whether MSCs from SLE patients were senescent and to determine the mechanism underlying this phenomenon. MSCs from both untreated and treated SLE patients showed characteristics of senescence. The expression of p16(INK4A) was significantly increased, whereas levels of CDK4, CDK6 and p-Rb expression were decreased in the MSCs from both untreated and treated SLE patients. Knockdown of p16(INK4A) expression reversed the senescent features of MSCs and upregulated TGF-β expression. In vitro, when purified CD4+ T cells were incubated with p16(INK4A)-silenced SLE MSCs, the percentage of regulatory T cells was significantly increased. Further, we have found that p16(INK4A) promotes MSC senescence via the suppression of the extracellular signal regulated kinase (ERK) pathway. p16(INK4A) knockdown up-regulated ERK1/2 activation. Our results demonstrated that MSCs from SLE patients were senescent and that p16 (INK4A) plays an essential role in the process by inhibiting ERK1/2 activation.

Topics: Adolescent; Adult; Bone Marrow Cells; Cells, Cultured; Cellular Senescence; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; Cyclin-Dependent Kinase Inhibitor p16; Female; Humans; Lupus Erythematosus, Systemic; Mesenchymal Stem Cells; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; RNA Interference; RNA, Small Interfering; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Up-Regulation; Young Adult

We aimed to compare bone-marrow-derived mesenchymal stem cells (BMMSCs) between systemic lupus erythematosus (SLE) and normal controls by means of cDNA microarray, immunohistochemistry, immunofluorescence, and immunoblotting. Our results showed there were a total of 1, 905 genes which were differentially expressed by BMMSCs derived from SLE patients, of which, 652 genes were upregulated and 1, 253 were downregulated. Gene ontology (GO) analysis showed that the majority of these genes were related to cell cycle and protein binding. Pathway analysis exhibited that differentially regulated signal pathways involved actin cytoskeleton, focal adhesion, tight junction, and TGF-β pathway. The high protein level of BMP-5 and low expression of Id-1 indicated that there might be dysregulation in BMP/TGF-β signaling pathway. The expression of Id-1 in SLE BMMSCs was reversely correlated with serum TNF-α levels. The protein level of cyclin E decreased in the cell cycling regulation pathway. Moreover, the MAPK signaling pathway was activated in BMMSCs from SLE patients via phosphorylation of ERK1/2 and SAPK/JNK. The actin distribution pattern of BMMSCs from SLE patients was also found disordered. Our results suggested that there were distinguished differences of BMMSCs between SLE patients and normal controls.

Topics: Actin Cytoskeleton; Adult; Aged; Bone Marrow Cells; Bone Morphogenetic Protein 5; Cells, Cultured; Cyclin E; Female; Focal Adhesions; Gene Expression Profiling; Humans; Inhibitor of Differentiation Protein 1; Lupus Erythematosus, Systemic; Male; MAP Kinase Signaling System; Mesenchymal Stem Cells; Middle Aged; Mitogen-Activated Protein Kinases; Phosphorylation; Tight Junctions; Transforming Growth Factor beta; Young Adult

T-lymphocytes are believed to play an important role in the pathogenesis of discoid lupus erythematosus (DLE). However, the reasons that lead to loss of tolerance and to development of autoimmunity in DLE remain unclear. In the present paper, we investigated serum levels of the regulatory cytokines transforming growth factor (TGF)-β and interleukin (IL)-10 in 25 newly diagnosed patients with DLE, 15 with systemic lupus erythematosus (SLE), 10 with psoriasis, 10 with atopic dermatitis (AD) and 20 healthy controls (HC). TGF-β serum levels were significantly lower in patients with DLE compared with patients with psoriasis and HC, while no differences were found between DLE, SLE and AD (medians: DLE: 28.49 ng/ml; psoriasis: 42.77 ng/ml; HC: 43.71 ng/ml; DLE vs. psoriasis: p < 0.05; DLE vs. HC: p < 0.05). IL-10 concentrations were reduced in DLE serum samples with respect to SLE, psoriasis, AD and HC (medians: DLE: 46.42 pg/ml; SLE: 127.64 pg/ml; psoriasis: 109.3 pg/ml; AD: 76.3 pg/ml; HC: 114.71 pg/ml; DLE vs. SLE: p < 0.05; DLE vs. psoriasis: p < 0.05; DLE vs. AD: p < 0.05; DLE vs. HC: p < 0.05). The downregulation of TGF-β and IL-10 in DLE may lead to defective immune suppression and thus to the generation of the tissue injury that is found in lupus patients.

Topics: Adult; Case-Control Studies; Dermatitis, Atopic; Down-Regulation; Female; Humans; Interleukin-10; Lupus Erythematosus, Discoid; Lupus Erythematosus, Systemic; Male; Middle Aged; Psoriasis; Transforming Growth Factor beta

Monocytes are a key component of the innate immune system involved in the regulation of the adaptive immune response. Previous studies have focused on apoptotic cell clearance abnormalities in systemic lupus erythematosus (SLE) monocytes. However, whether SLE monocytes might express unique patterns of cytokine secretion in response to apoptotic cells is still unknown. Here, we used monocytes from healthy controls and SLE patients to evaluate the production of TNF-α and TGF-β in response to apoptotic cells. Upon recognition of apoptotic material, monocytes from healthy controls showed prominent TGF-β secretion (mean ± SD: 824.6±144.3 pg/ml) and minimal TNF-α production (mean ± SD: 32.6±2.1 pg/ml). In contrast, monocytes from SLE patients had prominent TNF-α production (mean ± SD: 302.2±337.5 pg/ml) and diminished TGF-β secretion (mean ± SD: 685.9±615.9 pg/ml), a difference that was statistically significant compared to normal monocytes (p≤10(-6) for TNF-α secretion, and p = 0.0031 for TGF-β, respectively). Interestingly, the unique cytokine response by SLE monocytes was independent of their phagocytic clearance efficiency, opsonizing autoantibodies and disease activity. We further showed that nucleic acids from apoptotic cells play important role in the induction of TNF-α by lupus monocytes. Together, these observations suggest that, in addition to potential clearance defects, monocytes from SLE patients have an abnormal balance in the secretion of anti- and pro-inflammatory cytokines in response to apoptotic cells. Since the abnormal cytokine response to apoptotic material in SLE is not related to disease activity and opsonizing autoantibodies, it is possible that this response might be an intrinsic property of lupus monocytes. The studies focus attention on toll-like receptors (TLRs) and their downstream pathways as mediators of this response.

Topics: Adult; Apoptosis; Cell Survival; Deoxyribonucleases; Female; Humans; Inflammation Mediators; Jurkat Cells; Lupus Erythematosus, Systemic; Male; Middle Aged; Monocytes; Nucleic Acids; Phagocytosis; Ribonucleases; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The mechanisms underlying the loss of self-tolerance in systemic lupus erythematosus (SLE) are incompletely deciphered. TGF-β plays a key role in self-tolerance demonstrated by the onset of a fatal autoimmune syndrome associated with lupus autoantibodies in mice lacking a functional TGF-β receptor. The present work aims to define whether resistance to TGF-β might contribute to the pathogenesis of SLE.. Twenty-two patients with active SLE, 16 with other connective tissue diseases, and 10 healthy controls were prospectively included in this study. The effects of exogenous TGF-β1 on IL-2-dependent T-cell proliferation, IFN-γ secretion, and target gene transcription were analyzed on peripheral blood mononuclear cells.. Our results showed that 75% of patients with SLE or other connective tissue diseases were totally or partially resistant to the effects of TGF-β1. The responses to the anti-proliferative and transcriptional effects of TGF-β were, however, discordant in a high proportion of our patients. Hence, we distinguish three distinct profiles of resistance to TGF-β1 and suggest that patients may exhibit different defects affecting distinct points of TGF-β1 signaling pathways.. Our data demonstrate the presence of an impaired response of peripheral cells to TGF-β1 in patients with active SLE that may participate to the pathogenesis of the disease. Further studies will be necessary to delineate the mechanisms underlying the lymphocyte resistance to TGF-β1 in SLE.

Topics: Adolescent; Adult; Cell Proliferation; Female; Humans; Immune Tolerance; Interferon-gamma; Interleukin-2; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; Receptors, Transforming Growth Factor beta; Smad7 Protein; T-Lymphocytes; Transcription, Genetic; Transforming Growth Factor beta

In this work, we aimed to investigate the frequency, possible categories and clinical significance of circulating CD4+ ICOS+ FoxP3+ T cells in patients with systemic lupus erythematosus (SLE). The frequency of circulating CD4+ ICOS+ FoxP3+ T cells was analysed by flow-cytometric analysis in 32 SLE patients, 10 rheumatoid arthritis patients and 32 healthy controls. Production of IL-10 and mTGF-β by different CD4+ T-cell populations was determined by intracellular cytokine staining. Plasma levels of IL-10 and TGF-β were determined by enzyme-linked immunosorbent assay (ELISA). The frequency of circulating CD4+ ICOS+ FoxP3+ T cells was significantly increased in SLE patients as compared with control groups. The elevated frequency of CD4+ ICOS+ FoxP3+ T cells had a positive correlation with SLE Disease Activity Index (SLEDAI) scores and serum anti-dsDNA but a negative correlation with serum C3. Additionally, the CD4+ ICOS+ Foxp3+ T cells contained significantly higher percentages of IL-10-producing cells than CD4+ ICOS- Foxp3+ T cells. A significant positive correlation was also observed between the frequency of CD4+ ICOS+ Foxp3+ T cells and the plasma level of IL-10 in SLE patients. In conclusion, an increased frequency of circulating CD4+ ICOS+ Foxp3+ T cells was observed in patients with SLE, suggesting its potential importance in the immunopathogenesis of SLE. Analysis of the CD4+ ICOS+ FoxP3+ T-cell population may be useful for the evaluation of lupus disease severity.

Topics: Adult; Antigens, Differentiation, T-Lymphocyte; Arthritis, Rheumatoid; Case-Control Studies; CD4-Positive T-Lymphocytes; Cohort Studies; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Inducible T-Cell Co-Stimulator Protein; Interleukin-10; Lupus Erythematosus, Systemic; Male; Middle Aged; Severity of Illness Index; Transforming Growth Factor beta; Young Adult

Therapeutic agents currently in use to treat systemic lupus erythematosus (SLE) are predominantly immunosuppressive agents with limited specificities. Multiple groups, including ours, have illustrated that inducing tolerance in SLE animal models ameliorates disease symptoms and increases survival. We examined if oral administration of a tolerogenic peptide could affect SLE disease progression. The pConsensus (pCons) peptide, based on protein sequences of anti-double stranded (anti-ds)DNA antibodies, induces tolerance through upregulation of regulatory T cells when administered intravenously. Six different forms of pCons, including multiple antigenic peptides (MAP) and cyclic peptides made up of L- and D-amino acids, at three different concentrations, were fed to BWF1 SLE-susceptible mice for 30 weeks. Mice fed 100 µg of L-MAP or D-MAP had less cumulative proteinuria and serum anti-dsDNA antibody levels than controls. In addition, animals in these groups also survived significantly longer than controls with a corresponding increase in serum transforming growth factor beta (TGFβ, implying a protective role for pCons-induced regulatory T cells. Oral administration of a tolerogenic peptide is a safe, effective method for ameliorating SLE disease manifestations and prolonging survival in SLE-prone mice. Induction of oral tolerance using modified pCons peptides could lead to a novel targeted therapy for human SLE.

Topics: Administration, Oral; Animals; Antibodies, Antinuclear; Humans; Immune Tolerance; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Mice; Nephritis; Organic Chemicals; Peptides; Survival Rate; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

As a component of the innate immune cell population, γδ T cells are involved in tumor immunosurveillance and host defense against viral invasion. In this study, we demonstrated a novel function of human γδ T cells as regulatory cells by detecting their suppressive effect on the proliferation of autologous naive CD4(+) T cells. These regulatory γδ T cells (γδ Tregs) could be generated in vitro by stimulating with anti-TCRγδ in the presence of TGF-β and IL-2. Similar to CD4(+)Foxp3(+) Tregs, γδ Tregs also expressed Foxp3. Additionally, they primarily belonged to the Vδ1 subset with a CD27(+)CD25(high) phenotype. Furthermore, these γδ Tregs showed an immunoregulatory activity mainly through cell-to-cell contact. Importantly, this γδ regulatory population decreased in the peripheral blood of systemic lupus erythematosus patients, suggesting a potential mechanism in understanding the pathogenesis of systemic lupus erythematosus.

Topics: Adult; Aged; Blood Cells; Case-Control Studies; CD4-Positive T-Lymphocytes; Cell Proliferation; Cells, Cultured; Female; Humans; Immunity, Innate; Immunophenotyping; Lupus Erythematosus, Systemic; Lymphocyte Count; Male; Middle Aged; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Transforming growth factor β (TGFβ) plays a central role in maintaining immune homeostasis by regulating the initiation and termination of immune responses and thus preventing the development of autoimmune diseases. In this study, we describe an essential mechanism by which the actin regulatory protein Coronin 1A (Coro1A) ensures the proper response of Th17 CD4(+) T cells to TGFβ. Coro1A has been established as a key player in T cell survival, migration, activation, and Ca(2+) regulation in naive T cells. We show that mice lacking Coro1a developed less severe experimental autoimmune encephalomyelitis (EAE). Unexpectedly, upon the re-induction of EAE, Coro1a(-/-) mice exhibited enhanced EAE signs that correlated with increased numbers of IL-17 producing CD4(+) cells in the central nervous system (CNS) compared to wild-type mice. In vitro differentiated Coro1a(-/-) Th17 CD4(+) T cells consistently produced more IL-17 than wild-type cells and displayed a Th17/Th1-like phenotype in regard to the expression of the Th1 markers T-bet and IFNγ. Mechanistically, the Coro1a(-/-) Th17 cell phenotype correlated with a severe defect in TGFβR-mediated SMAD3 activation. Taken together, these data provide experimental evidence of a non-redundant role of Coro1A in the regulation of Th17 CD4(+) cell effector functions and, subsequently, in the development of autoimmunity.

Topics: Animals; Cell Differentiation; Cell Movement; Disease Models, Animal; Female; Flow Cytometry; Gene Expression; Interferon-gamma; Interleukin-17; Lupus Erythematosus, Systemic; Mice; Mice, Knockout; Microfilament Proteins; Real-Time Polymerase Chain Reaction; Receptors, Transforming Growth Factor beta; Signal Transduction; Smad3 Protein; Th1 Cells; Th17 Cells; Transforming Growth Factor beta

CD25(+) FOXP3(+) CD4(+) regulatory T cells (Tregs) are induced by transforming growth factor β (TGFβ) and further expanded by retinoic acid (RA). We have previously shown that this process was defective in T cells from lupus-prone mice expressing the novel isoform of the Pbx1 gene, Pbx1-d. This study tested the hypothesis that CD4(+) T cells from systemic lupus erythematosus (SLE) patients exhibited similar defects in Treg induction in response to TGFβ and RA, and that PBX1-d expression is associated with this defect.. Peripheral blood mononuclear cells (PBMCs) were collected from 142 SLE patients and 83 healthy controls (HCs). The frequency of total, memory and naïve CD4(+) T cells was measured by flow cytometry on fresh cells. PBX1 isoform expression in purified CD4(+) T cells was determined by reverse transcription polymerase chain reaction (RT-PCR). PBMCs were stimulated for three days with anti-CD3 and anti-CD28 in the presence or absence of TGFβ and RA. The expression of CD25 and FOXP3 on CD4(+) T cells was then determined by flow cytometry. In vitro suppression assays were performed with sorted CD25(+) and CD25(-) FOXP3(+) T cells. CD4(+) T cell subsets or their expansion were compared between patients and HCs with two-tailed Mann-Whitney tests and correlations between the frequencies of two subsets were tested with Spearman tests.. The percentage of CD25(-) FOXP3(+) CD4(+) (CD25(-) Tregs) T cells was greater in SLE patients than in HCs, but these cells, contrary to their matched CD25(+) counterparts, did not show a suppressive activity. RA-expansion of TGFβ-induced CD25(+) Tregs was significantly lower in SLE patients than in HCs, although SLE Tregs expanded significantly more than HCs in response to either RA or TGFβ alone. Defective responses were also observed for the SLE CD25(-) Tregs and CD25(+) FOXP3(-) activated CD4(+) T cells as compared to controls. PBX1-d expression did not affect Treg induction, but it significantly reduced the expansion of CD25- Tregs and prevented the reduction of the activated CD25(+) FOXP3(-) CD4(+) T cell subset by the combination of TGFβ and RA.. We demonstrated that the induction of Tregs by TGFβ and RA was defective in SLE patients and that PBX1-d expression in CD4(+) T cells is associated with an impaired regulation of FOXP3 and CD25 by TGFβ and RA on these cells. These results suggest an impaired integration of the TGFβ and RA signals in SLE T cells and implicate the PBX1 gene in this process.

Topics: Adult; Aged; Antineoplastic Agents; CD4-Positive T-Lymphocytes; Cell Division; Cells, Cultured; Female; Flow Cytometry; Forkhead Transcription Factors; Humans; Immunologic Memory; Interleukin-2 Receptor alpha Subunit; Interleukin-7 Receptor alpha Subunit; Leukocyte Common Antigens; Lupus Erythematosus, Systemic; Male; Middle Aged; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Tretinoin; Young Adult

To investigate the association between deficiencies of early components in the classical complement pathway and the development of SLE.. Forty inbred C57BL/6J mice and 40 knockout C4 complement gene (C4KO) mice, which included 10 mice in each age group (2, 4, 6, and 8 months) were used. The enumeration of CD4+CD25+ Tregs frequencies in bone marrow, spleen and peripheral blood from both normal and C4KO groups were performed by flow cytometry. The expression levels of Foxp3 and TGF-beta in the same tested tissues were measured using real time PCR. The antinuclear antibodies (ANA) were semi-quantitatively measured using ELISA.. We report decreased frequencies of CD4+CD25+ Tregs and reduced expression levels of Foxp3 and TGF-beta, which efficiently program the development and function of Tregs, in lymphoid tissues and peripheral blood of C4KO mice. In this study, C4KO mice have higher titers of ANA than those of normal mice. Higher frequencies of mice positive for ANA are also found in older mice.. The deficiency of the C4 gene induces the decreased numbers of Tregs that further increase the production of ANA resulting in the development of an autoimmune disorder. The outcomes of our study help us to understand the association between the deficiency of C4 in the classical complement pathway and development of autoimmune disorder via the role of Tregs.

Topics: Animals; Antibodies, Antinuclear; Bone Marrow; Complement C4; Forkhead Transcription Factors; Interleukin-2 Receptor alpha Subunit; Lupus Erythematosus, Systemic; Mice; Mice, Inbred C57BL; Mice, Knockout; Spleen; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Dysregulated expression of Bcl-xL and Bcl-2 may initiate the development of autoimmune diseases including systemic lupus erythematosus (SLE). A tolerogenic peptide designated hCDR1 was shown to ameliorate manifestations of spontaneous and induced murine SLE. Recently, we demonstrated that Bcl-xL plays a critical role in the modulating effects of hCDR1, as manifested by reducing the state of activation of lymphocytes and by down-regulating the secretion of the pathogenic cytokines, IFN-gamma and IL-10. Here we studied the role of Bcl-xL in the development and function of CD4 regulatory T-cells (Treg) from hCDR1-treated, SLE-afflicted (New-Zealand-Black x New-Zealand-White) F1 mice. We report that Bcl-xL was up-regulated in CD4 Treg of tolerized mice, where it played a role in inducing the regulatory/inhibitory molecules Foxp3, CTLA-4, and TGF-beta and in repressing PD-1. Further, Bcl-xL mediated the induction of CTLA-4 and TGF-beta in effector T cells (Teff) by CD4 Treg of the tolerized mice. The induction of Bcl-xL in Teff by Treg was TGF-beta dependent and CTLA-4 independent, leading to inhibition of proliferation and to a decrease in activated Teff. We conclude that Bcl-xL is required for the development and function of CD4 Treg, which ameliorate lupus following treatment with a tolerogenic peptide.

Topics: Animals; Antibodies, Monoclonal; Antigens, CD; bcl-X Protein; Cell Proliferation; Cells, Cultured; Female; Forkhead Transcription Factors; Humans; Immune Tolerance; Interferon-gamma; Interleukin-10; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Peptide Fragments; Proto-Oncogene Proteins c-bcl-2; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Up-Regulation

To compare frequency and functional activity of peripheral blood (PB) Th(c)17, Th(c)1 and Treg cells and the amount of type 2 cytokines mRNA we recruited SLE patients in active (n=15) and inactive disease (n=19) and healthy age- and gender-matched controls (n=15). The study of Th(c)17, Th(c)1 and Treg cells was done by flow cytometry and cytokine mRNA by real-time PCR. Compared to NC, SLE patients present an increased proportion of Th(c)17 cells, but with lower amounts of IL-17 per cell and also a decreased frequency of Treg, but with increased production of TGF-beta and FoxP3 mRNA. Iotan active compared to inactive SLE, there is a marked decreased in frequency of Th(c)1 cells, an increased production of type 2 cytokines mRNA and a distinct functional profile of Th(c)17 cells. Our findings suggest a functional disequilibrium of T-cell subsets in SLE which may contribute to the inflammatory process and disease pathogenesis.

Topics: Adult; Antigens, CD; Cell Count; Cell Separation; Female; Flow Cytometry; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Interleukin-17; Lupus Erythematosus, Systemic; Male; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocyte Subsets; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Regulatory; Th2 Cells; Transforming Growth Factor beta

Cytokines are central regulators of the immune response but the workings of this complex network in systemic lupus erythematosus (SLE) are not fully understood. We investigated a range of inflammatory and immune-modulating cytokines to determine their value as biomarkers for disease subsets in SLE.. This was a cross-sectional study in 102 patients with SLE (87% women, disease duration 10.6 yrs). Circulating concentrations of interleukin 1β (IL-1β), IL-4, IL-6, IL-10, IL-12, IL-17, monocyte chemotactic protein 1 (MCP-1), macrophage inflammatory protein 1 (MIP-1α), MIP-1β, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and total transforming growth factor-β1 (TGF-β1) were related to disease activity (SLE Disease Activity Index; SLEDAI), lymphocyte subsets, autoantibody levels, accrued damage (Systemic Lupus International Collaborating Clinics/ACR Damage Index; SDI), and concomitant treatment.. Patients with SLE had lower levels of TGF-β1 (p = 0.01) and IL-1β (p = 0.0004) compared to controls. TGF-β1 levels were lower in patients with SLEDAI scores 1-10 and SDI > 3; and were correlated with CD4+, CD8+, and natural killer cell counts; and were independent of steroid or cytotoxic drug use. Treatment with cardiovascular drugs was associated with lower IL-12 levels. No consistent disease associations existed for the other cytokines investigated.. Lower TGF-β1 was the most consistent cytokine abnormality in patients with SLE. The associations with disease activity, lymphocyte subsets, and damage suggest that TGF-β1 may be a therapeutic target of interest in SLE.

Topics: Adult; Aged; Biomarkers; Chemokine CCL2; Cross-Sectional Studies; Female; Humans; Interferon-gamma; Interleukins; Lupus Erythematosus, Systemic; Macrophage Inflammatory Proteins; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Animals; Autoimmunity; Humans; Interleukins; Lupus Erythematosus, Systemic; T-Lymphocytes; Transforming Growth Factor beta

The programmed death-1 (PD-1)/programmed death-1 ligand 1 (PD-L1) pathway regulates both stimulatory and inhibitory signals. In some conditions, PD-1/PD-L1 inhibits T and B cell activation, induces anergy, and reduces cytotoxicity in CD8(+) T cells. In other conditions, PD-l/PD-L1 has costimulatory effects on T cells. We recently showed that induction of suppressive CD8(+)Foxp3(+) T cells by immune tolerance of lupus-prone (New Zealand black × New Zealand white)F(1) (BWF(1)) mice with the anti-DNA Ig-based peptide pConsensus (pCons) is associated with significantly reduced PD-1 expression on those cells. In this study, we tested directly the role of PD-1 by administering in vivo neutralizing Ab to PD-1 to premorbid BWF(1) and healthy control mice. Anti-PD-1-treated mice were protected from the onset of lupus nephritis for 10 wk, with significantly improved survival. Although the numbers of T cells declined in aging control mice, they were maintained in anti-PD-1-treated mice, including CD8(+)Foxp3(+) T cells that suppressed syngeneic CD4(+)CD25(-) T cell proliferation and IFN-γ production, reduced production of IgG and anti-dsDNA IgG, induced apoptosis in syngeneic B cells, and increased IL-2 and TGF-β production. The administration of anti-PD-1 Ab to BWF(1) mice after induction of tolerance with pCons abrogated tolerance; mice developed autoantibodies and nephritis at the same time as control mice, being unable to induce CD8(+)Foxp3(+) T suppressor cells. These data suggest that tightly regulated PD-1 expression is essential for the maintenance of immune tolerance mediated by those CD8(+)Foxp3(+) T cells that suppress both T(h) cells and pathogenic B cells. PD-1 regulation could represent a target to preserve tolerance and prevent autoimmunity.

Topics: Aging; Amino Acid Sequence; Animals; Antibodies, Blocking; Antigens, Surface; Apoptosis; Apoptosis Regulatory Proteins; Autoantibodies; CD8-Positive T-Lymphocytes; Crosses, Genetic; Down-Regulation; Female; Immune Tolerance; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Inbred NZB; Programmed Cell Death 1 Receptor; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is characterized in its early stages by the expansion of autoreactive T cells that trigger B-cell activation with subsequent multi-organ injury. Dendritic cells (DCs) in lupus were found to display an aberrant phenotype with higher expression of the maturation markers major histocompatibility complex (MHC) class II, CD80 and CD86, as well as higher production of proinflammatory cytokines including interleukin-12 (IL-12), resulting in an increased ability to activate T cells. A peptide (hCDR1) based on the complementarity determining region-1 of an anti-DNA antibody ameliorated SLE in both induced and spontaneous lupus models by downregulating T-cell functions. Our objectives were to determine whether DCs play a role in promoting the beneficial effects of hCDR1. We showed here that treatment with hCDR1 lowered the expression levels of MHC class II, CD80 and CD86 on DCs. The latter effect was associated with downregulation of messenger RNA expression and secretion of IL-12, a cytokine that upregulated T-cell proliferation and interferon-gamma (IFN-gamma) secretion. Moreover, DCs derived from hCDR1-treated mice downregulated proliferation and IFN-gamma secretion by T cells from untreated mice. Upregulation of transforming growth factor-beta (TGF-beta) secretion by T cells, following treatment with hCDR1, resulted in downregulation of IFN-gamma production and contributed to the phenotypic changes and magnitude of IL-12 secretion by DCs. The ameliorating effects of hCDR1 are therefore mediated at least partially by the upregulated secretion of TGF-beta by T cells that contribute to the induction of DCs with immature phenotype and suppressed functions. The resulting DCs further downregulate autoreactive T-cell functions.

Topics: Animals; Autoantigens; B7-1 Antigen; B7-2 Antigen; Dendritic Cells; Down-Regulation; Female; Histocompatibility Antigens Class II; Humans; Interferon-gamma; Interleukin-12; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Nerve Tissue Proteins; Peptides; T-Lymphocytes; Transforming Growth Factor beta; Up-Regulation

CD4+CD25+ regulatory T cells play an essential role in maintaining immune homeostasis and preventing autoimmunity. Therefore, defects in Treg development, maintenance or function have been associated with several human autoimmune diseases including Systemic Lupus Erythematosus (SLE), a systemic autoimmune disease characterized by loss of tolerance to nuclear components and significantly more frequent in females.. To investigate the involvement of Treg in SLE pathogenesis, we determined the frequency of CD4+CD25+CD45RO+ T cells, which encompass the majority of Treg activity, in the PBMC of 148 SLE patients (76 patients were part of 54 families), 166 relatives and 117 controls. SLE patients and their relatives were recruited in several Portuguese hospitals and through the Portuguese Lupus Association. Control individuals were blood donors recruited from several regional blood donor centers. Treg frequency was significantly lower in SLE patients than healthy controls (z = -6.161, P < 0.00001) and intermediate in the relatives' group. Remarkably, this T cell subset was also lower in females, most strikingly in the control population (z = 4.121, P < 0.001). We further ascertained that the decreased frequency of Treg in SLE patients resulted from the specific reduction of bona fide FOXP3+CD4+CD25+ Treg. Treg frequency was negatively correlated with SLE activity index (SLEDAI) and titers of serum anti-dsDNA antibodies. Both Treg frequency and disease activity were modulated by IVIg treatment in a documented SLE case. The segregation of Treg frequency within the SLE families was indicative of a genetic trait. Candidate gene analysis revealed that specific variants of CTLA4 and TGFbeta were associated with the decreased frequency of Treg in PBMC, while FOXP3 gene variants were associated with affection status, but not with Treg frequency.. SLE patients have impaired Treg production or maintenance, a trait strongly associated with SLE disease activity and autoantibody titers, and possibly resulting from the inability to convert FOXP3+CD25- into FOXP3+CD25+ T cells. Treg frequency is highly heritable within SLE families, with specific variants of the CTLA4 and TGFbeta genes contributing to this trait, while FOXP3 contributes to SLE through mechanisms not involving a modulation of Treg frequency. These findings establish that the genetic components in SLE pathogenesis include genes related to Treg generation or maintenance.

Topics: Age Factors; Antibodies, Antinuclear; Antigens, CD; CD4 Antigens; CTLA-4 Antigen; Female; Forkhead Transcription Factors; Genotype; Humans; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Leukocyte Common Antigens; Lupus Erythematosus, Systemic; Male; Polymorphism, Genetic; Quantitative Trait, Heritable; Severity of Illness Index; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

The NKG2D receptor stimulates natural killer cell and T cell responses upon engagement of ligands associated with malignancies and certain autoimmune diseases. However, conditions of persistent NKG2D ligand expression can lead to immunosuppression. In cancer patients, tumor expression and shedding of the MHC class I-related chain A (MICA) ligand of NKG2D drives proliferative expansions of NKG2D(+)CD4(+) T cells that produce interleukin-10 (IL-10) and transforming growth factor-beta, as well as Fas ligand, which inhibits bystander T cell proliferation in vitro. Here, we show that increased frequencies of functionally equivalent NKG2D(+)CD4(+) T cells are inversely correlated with disease activity in juvenile-onset systemic lupus erythematosus (SLE), suggesting that these T cells may have regulatory effects. The NKG2D(+)CD4(+) T cells correspond to a normally occurring small CD4 T cell subset that is autoreactive, primed to produce IL-10, and clearly distinct from proinflammatory and cytolytic CD4 T cells with cytokine-induced NKG2D expression that occur in rheumatoid arthritis and Crohn's disease. As classical regulatory T cell functions are typically impaired in SLE, it may be clinically significant that the immunosuppressive NKG2D(+)CD4(+) T cells appear functionally uncompromised in this disease.

Topics: Age of Onset; Antigens, CD; CD4 Antigens; CD4-Positive T-Lymphocytes; Cytokines; Fetal Blood; Humans; Immunologic Memory; Infant, Newborn; Interleukin-10; Killer Cells, Natural; Lupus Erythematosus, Systemic; NK Cell Lectin-Like Receptor Subfamily K; Reference Values; T-Lymphocyte Subsets; T-Lymphocytes; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is an autoimmune disorder characterized by dysregulation of cytokines, apoptosis, and B- and T-cell functions. The tolerogenic peptide, hCDR1 (Edratide), ameliorated the clinical manifestations of murine lupus via down-regulation of pro-inflammatory cytokines and apoptosis, up-regulation of the immunosuppressive cytokine TGF-beta, and the induction of regulatory T-cells. In the present study, gene expression was determined in peripheral blood mononuclear cells of 9 lupus patients that were treated for 26 weeks with either hCDR1 (five patients), or placebo (four patients). Disease activity was assessed by SLEDAI-2K and the BILAG scores. Treatment with hCDR1 significantly down-regulated the mRNA expression of the pathogenic cytokines IL-1beta, TNF-alpha, IFN-gamma, and IL-10, of BLyS (B-lymphocyte stimulator) and of the pro-apoptotic molecules caspase-3 and caspase-8. In contrast, the treatment up-regulated in vivo gene expression of both TGF-beta and FoxP3. Furthermore, hCDR1 treatment resulted in a significant decrease in SLEDAI-2K (from 8.0+/-2.45 to 4.4+/-1.67; P=0.02) and BILAG (from 8.2+/-2.7 to 3.6+/-2.9; P=0.03) scores. Thus, the tolerogenic peptide hCDR1, immunomodulates, in vivo, the expression of genes that play a role in SLE, consequently restoring the global immune dysregulation of lupus patients. Hence, hCDR1 has a potential role as a novel disease-specific treatment for lupus patients.

Topics: Adult; Antibodies, Monoclonal; Apoptosis; Caspases; Complementarity Determining Regions; Cytokines; Disease Progression; Female; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Immunotherapy; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Male; Middle Aged; Peptide Fragments; Self Tolerance; Severity of Illness Index; Transforming Growth Factor beta; Treatment Outcome

Systemic lupus erythematosus (SLE) is an autoimmune disease mediated by T and B cells. It is characterized by a variety of autoantibodies and systemic clinical manifestations. A tolerogenic peptide, designated hCDR1, ameliorated the serological and clinical manifestations of SLE in both spontaneous and induced models of lupus. In the present study, we evaluated the status of mature B cells in the bone marrow (BM) of SLE-afflicted mice, and determined the effect of treatment with the tolerogenic peptide hCDR1 on these cells. We demonstrate herein that mature B cells of the BM of SLE-afflicted (New Zealand Black x New Zealand White)F(1) mice were largely expanded, and that treatment with hCDR1 down-regulated this population. Moreover, treatment with hCDR1 inhibited the expression of the pathogenic cytokines [interferon-gamma and interleukin (IL)-10], whereas it up-regulated the expression of transforming growth factor-beta in the BM. Treatment with hCDR1 up-regulated the rates of apoptosis of mature B cells. The latter was associated with inhibited expression of the survival Bcl-xL gene and of IL-7 by BM cells. Furthermore, the addition of recombinant IL-7 abrogated the suppressive effects of hCDR1 on Bcl-xL in the BM cells and resulted in elevated levels of apoptosis. Hence, the down-regulated production of IL-7 contributes to the hCDR1-mediated apoptosis of mature B cells in the BM of SLE-afflicted mice.

Topics: Animals; Antibodies, Monoclonal; Apoptosis; B-Lymphocyte Subsets; Bone Marrow Cells; Disease Models, Animal; Down-Regulation; Female; Immune Tolerance; Interferon-gamma; Interleukin-10; Interleukin-7; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; Peptide Fragments; Reverse Transcriptase Polymerase Chain Reaction; Spleen; Transforming Growth Factor beta; Up-Regulation

Compared with conventional drug therapy, autologous hemopoietic stem cell transplantation (HSCT) can induce very-long-term remission in refractory lupus patients. Herein, we show that in posttransplant patients, both CD4(+)CD25(high)FoxP3(+) and an unusual CD8(+)FoxP3(+) Treg subset return to levels seen in normal subjects; accompanied by almost complete inhibition of pathogenic T cell response to critical peptide autoepitopes from histones in nucleosomes, the major lupus autoantigen from apoptotic cells. In addition to a stably sustained elevation of FoxP3, posttransplant CD8 T cells also maintained markedly higher expression levels of latency-associated peptide (LAP), CD103, PD-1, PD-L1, and CTLA-4, as compared with pretransplant CD8 T cells that were identically treated by a one-time activation and rest in short-term culture. The posttransplant CD8 regulatory T cells (Treg) have autoantigen-specific and nonspecific suppressive activity, which is contact independent and predominantly TGF-beta dependent. By contrast, the pretransplant CD8 T cells have helper activity, which is cell contact dependent. Although CD4(+)CD25(high) Treg cells return during clinical remission of conventional drug-treated lupus, the posttransplant patient's CD8 Treg cells are considerably more potent, and they are absent in drug-treated patients in whom CD4 T cell autoreactivity to nucleosomal epitopes persists even during clinical remission. Therefore, unlike conventional drug therapy, hemopoietic stem cell transplantation generates a newly differentiated population of LAP(high)CD103(high) CD8(TGF-beta) Treg cells, which repairs the Treg deficiency in human lupus to maintain patients in true immunological remission.

Topics: Adolescent; Adult; Antigens, CD; Apoptosis Regulatory Proteins; B7-H1 Antigen; CD8-Positive T-Lymphocytes; CTLA-4 Antigen; Female; Hematopoietic Stem Cell Transplantation; Humans; Integrin alpha Chains; Interferon-gamma; Interleukin-13; Lupus Erythematosus, Systemic; Male; Middle Aged; Nucleosomes; Programmed Cell Death 1 Receptor; Remission, Spontaneous; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Young Adult

Most researchers believe that the peroxisome proliferative activated receptor gamma (PPARgamma-2) and bone morphogenetic protein receptor type II (BMPR2) play important roles in steroid-induced osteonecrosis (ON). However, the molecular mechanism of this process is still unclear. Recent studies indicate that steroid treatments cause adipocyte formation due to differentiation of mesenchymal stem cells, which then prevents osteoblast formation. This study examined PPARgamma-2, bone morphogenetic protein 2 (BMP2), and BMPR2 in patients with systemic lupus erythromatosus (SLE) who eventually developed ON after prolonged steroid treatment. The subjects of this experiment included 220 SLE patients who had undergone steroid treatment for at least 2 years. Fifty-five of the 220 patients were ON patients, and 165 were non-ON patients. Real-time PCR was performed to analyze the expression of the PPARgamma-2, BMP2, and BMPR2 mRNA in the peripheral blood of these patients. The results indicated that the expression of PPARgamma-2 mRNA increased 37% in the ON patients' peripheral blood, but the expression of BMPR2 mRNA decreased 57%. The average expression of the PPARgamma-2 mRNA in the ON patients was significantly higher than that in the non-ON patients (p = 0.044). Conversely, the expression of BMPR2 mRNA was significantly lower than that in non-ON patients (p = 0.036), but the expression of BMP2 mRNA did not significantly differ. This study demonstrated that the PPARgamma-2 and BMPR2 have important roles in the ON process after prolonged steroid administration in SLE patients; however, the detailed molecular mechanisms of this process require further study.

Topics: Adrenal Cortex Hormones; Adult; Bone Morphogenetic Protein 2; Bone Morphogenetic Protein Receptors, Type II; Bone Morphogenetic Proteins; Female; Gene Expression; Humans; Immunosuppressive Agents; Lupus Erythematosus, Systemic; Male; Middle Aged; Osteonecrosis; PPAR gamma; Transforming Growth Factor beta

Ample evidence suggests a role of TGF-beta in preventing autoimmunity. Multiorgan inflammatory disease, spontaneous activation of self-reactive T cells, and autoantibody production are hallmarks of autoimmune diseases, such as lupus. These features are reminiscent of the immunopathology manifest in TGF-beta1-deficient mice. In this study, we show that lupus-prone (New Zealand Black and White)F(1) mice have reduced expression of TGF-beta1 in lymphoid tissues, and TGF-beta1 or TGF-beta1-producing T cells suppress autoantibody production. In contrast, the expression of TGF-beta1 protein and mRNA and TGF-beta signaling proteins (TGF-beta receptor type II and phosphorylated SMAD3) increases in the target organs, i.e., kidneys, of these mice as they age and develop progressive organ damage. In fact, the levels of TGF-beta1 in kidney tissue and urine correlate with the extent of chronic lesions that represent local tissue fibrosis. In vivo TGF-beta blockade by treatment of these mice with an anti-TGF-beta Ab selectively inhibits chronic fibrotic lesions without affecting autoantibody production and the inflammatory component of tissue injury. Thus, TGF-beta plays a dual, seemingly paradoxical, role in the development of organ damage in multiorgan autoimmune diseases. According to our working model, reduced TGF-beta in immune cells predisposes to immune dysregulation and autoantibody production, which causes tissue inflammation that triggers the production of anti-inflammatory cytokines such as TGF-beta in target organs to counter inflammation. Enhanced TGF-beta in target organs, in turn, can lead to dysregulated tissue repair, progressive fibrogenesis, and eventual end-organ damage.

Topics: Animals; Autoantibodies; Autoimmune Diseases; Fibrosis; Kidney; Lupus Erythematosus, Systemic; Lymphoid Tissue; Mice; Mice, Mutant Strains; Signal Transduction; Transforming Growth Factor beta

Systemic lupus erythematosus is an autoimmune disease caused by autoantibodies, including IgG anti-DNA. New Zealand Black/New Zealand White F(1) female mice, a model of spontaneous polygenic systemic lupus erythematosus, tolerized with an artificial peptide (pConsensus) based on anti-DNA IgG sequences containing MHC class I and class II T cell determinants, develop regulatory CD4+CD25+ T cells and CD8+ inhibitory T cells (CD8+ Ti), both of which suppress autoantibody production. CD8+ Ti inhibit primarily via secretion of TGF-beta. In the present study, we show that the inhibitory function of CD8+ T cells from tolerized mice is sustained for up to 8 wk and at all times depends on expression of Foxp3. Both CD28-positive and CD28-negative CD8+ T cells contain inhibitory cells, but the expression of mRNA for Foxp3 and for TGF-beta is higher and lasts longer in the CD28- subset. In vitro addition of TGF-beta (in the presence of IL-2) induces Foxp3 expression in a dose-response manner. Gene inhibition or blockade with small interfering RNA of Foxp3 abrogates the ability of the CD8+ Ti to inhibit anti-DNA production and the proliferation of CD4+ Th cells. Moreover, a significant correlation between expression of Foxp3 and ability of CD8+ Ti to secrete TGF-beta is observed. Therefore, CD8+ Ti in this system of tolerance are similar to CD4+CD25+ regulatory T cells in their dependence on expression of Foxp3, and there may be a bidirectional Foxp3/TGF-beta autocrine loop that determines the ability of the CD8+ T cells to control autoimmunity.

Topics: Animals; Antibodies, Antinuclear; Autoimmunity; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Disease Models, Animal; Female; Forkhead Transcription Factors; Immune Tolerance; Immunoglobulin G; Lupus Erythematosus, Systemic; Mice; Mice, Inbred Strains; Peptides; RNA, Messenger; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

Subnanomolar doses of an unaltered, naturally occurring nucleosomal histone peptide epitope, H4(71-94), when injected s.c. into lupus-prone mice, markedly prolong lifespan by generating CD4+25+ and CD8+ regulatory T cells (Treg) producing TGF-beta. The induced Treg cells suppress nuclear autoantigen-specific Th and B cells and block renal inflammation. Splenic dendritic cells (DC) captured the s.c.-injected H4(71-94) peptide rapidly and expressed a tolerogenic phenotype. The DC of the tolerized animal, especially plasmacytoid DC, produced increased amounts of TGF-beta, but diminished IL-6 on stimulation via the TLR-9 pathway by nucleosome autoantigen and other ligands; and those plasmacytoid DC blocked lupus autoimmune disease by simultaneously inducing autoantigen-specific Treg and suppressing inflammatory Th17 cells that infiltrated the kidneys of untreated lupus mice. Low-dose tolerance with H4(71-94) was effective even though the lupus immune system is spontaneously preprimed to react to the autoepitope. Thus, H4(71-94) peptide tolerance therapy that preferentially targets pathogenic autoimmune cells could spare lupus patients from chronically receiving toxic agents or global immunosuppressants and maintain remission by restoring autoantigen-specific Treg cells.

Topics: Adoptive Transfer; Animals; Autoantibodies; Autoantigens; Autoimmune Diseases; B-Lymphocytes; CD4 Antigens; CD8 Antigens; Dendritic Cells; Forkhead Transcription Factors; Histones; Immune Tolerance; Immunoglobulin G; Immunosuppression Therapy; Interleukin-17; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Lupus Erythematosus, Systemic; Mice; Nucleosomes; Peptide Fragments; Peptides; T-Lymphocytes, Regulatory; Toll-Like Receptor 9; Transforming Growth Factor beta

Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine that mainly acts as an inhibitor of immune functions. A lack of functional TGF-beta leads to autoimmune disease in animal models and dysregulated TGF-beta signaling is implicated in human autoimmune diseases. To define target genes that play a part in the inhibitory role of TGF-beta in the immune system, we have identified genes stimulated by TGF-beta in macrophages by gene-chip analysis. One of the TGF-beta regulated genes is carboxypeptidase D (CpD), a 180-kDa type I membrane protein. We have demonstrated that CpD is regulated by TGF-beta in various cell types of both, murine and human origin and, interestingly, is significantly downregulated in CD14 positive cells isolated from patients with lupus erythematosus (LE). Moreover, we show that downregulation of CpD leads to downmodulation of TGF-beta itself, suggesting a role for CpD in a positive feedback loop, providing further evidence for a role of this enzyme in LE. To our knowledge, this is the first report that demonstrates carboxypeptidase D as a TGF-beta target gene that is implicated in the pathogenesis of LE.

Topics: Adult; Animals; Carboxypeptidases; Cell Line; Cell Line, Tumor; Down-Regulation; Enzyme Induction; Feedback, Physiological; Female; Gene Expression Regulation, Enzymologic; Humans; Lipopolysaccharide Receptors; Lupus Erythematosus, Systemic; Macrophages; Male; Mice; Mice, Inbred C57BL; Middle Aged; Signal Transduction; Transforming Growth Factor beta; Up-Regulation

To study the in vitro modulation of autogeneic and allogenic regulatory T lymphocytes proliferation by bone marrow mesenchymal stem cells (MSCs) in patients with systemic lupus erythematosis (SLE).. Human MSCs were separated with Percoll (1.073 g/ml) from bone marrow of patients with SLE or healthy subjects. The purity of MSCs was identified with the phenotypes by fluorescence active cell sorter (FACS). The subset regulatory T cells in the peripheral blood of patients with SLE were isolated with magnetic activated cell sorting (MACS) CD4 and CD25 microbeads. Lymphocytes or CD4+ CD25+ T cells isolated from the peripheral blood of SLE patients were cocultured either with autologous MSCs or MSCs from healty donors. The proliferation activities of lymphocytes and CD4+ CD25+ T cells were investigated with methyl thiazolyl tetrazolium( MTr) test. The level of IL-10 and transforming growth factor beta (TGFbeta) was determined with enzyme-linked immunosorbent assay (ELISA).. The lymphocyte activity in SLE was suppressed by autogeneic and allogeneic MSCs and the inhibition rate was 56.32% and 65.46%, respectively. A stronger immunosuppressive effect of allogeneic MSCs was detected. MSCs were capable of increasing the proportion of allogeneic and autogeneic regulatory T cells in a dose dependent fashion. MSCs stimulated CD4+ CD25+ T cells to produce IL-10 and TGFbeta.. MSCs can suppress lymphocyte proliferation and increase CD4+ CD25+ T cells. MSCs might play important roles in immunosuppressant lymphocyte proliferation and be important to cooperate with autogeneic hematopoietic stem cells in transplantation.

Topics: Adolescent; Adult; Cell Proliferation; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-10; Lewis X Antigen; Lupus Erythematosus, Systemic; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Middle Aged; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Apoptotic cells are considered an important auto-antigenic source in diseases such as systemic lupus erythematosus (SLE). A human monoclonal antibody demonstrating exquisite specificity towards late-stage apoptotic cells was generated from an SLE patient. Polyreactive recognition of ribonucleoproteins Ro52 and Ro60 was observed. The antibody significantly diminished the phagocytosis of apoptotic cells and a concomitant decrease in transforming growth factor-beta (TGF-beta) secretion was observed. Light and heavy chain sequencing revealed the antibody to be in essentially germline configuration. Elicited anti-idiotypic antibodies bound distinct self-antigens and showed augmented reactivity towards apoptotic cells as well. Thus, near-germline encoded antibodies recognizing antigens externalized during the process of apoptosis can mediate a variety of potentially pathogenic effects; decreases in the phagocytic uptake of dying cells would constitute a disease-perpetuating event and stimulation of the idiotypic network could lead to intermolecular epitope spreading, increasing the range of molecular targets..

Topics: Antibodies, Monoclonal; Antibody Specificity; Apoptosis; Autoantibodies; Autoantigens; Base Sequence; Cell Line; DNA; Epitopes; Humans; Immunoglobulin Idiotypes; Jurkat Cells; Lupus Erythematosus, Systemic; Phagocytosis; Transforming Growth Factor beta

The efficiency of activating latent transforming growth factor (TGF)-beta1 in systemic lupus erythematosus (SLE) may control the balance between inflammation and fibrosis, modulating the disease phenotype. To test this hypothesis we studied the ability to activate TGF-beta1 in SLE patients and control individuals within the context of inflammatory disease activity, cumulative organ damage and early atherosclerosis. An Activation Index (AI) for TGF-beta1 was determined for 32 patients with SLE and 33 age-matched and sex-matched control individuals by quantifying the increase in active TGF-beta1 under controlled standard conditions. Apoptosis in peripheral blood mononuclear cells was determined by fluorescence-activated cell sorting. Carotid artery intima-media thickness was measured using standard Doppler ultrasound. These measures were compared between patients and control individuals. In an analysis conducted in patients, we assessed the associations of these measures with SLE phenotype, including early atherosclerosis. Both intima-media thickness and TGF-beta1 AI for SLE patients were within the normal range. There was a significant inverse association between TGF-beta1 AI and levels of apoptosis in peripheral blood mononuclear cells after 24 hours in culture for both SLE patients and control individuals. Only in SLE patients was there a significant negative correlation between TGF-beta1 AI and low-density lipoprotein cholesterol (r = -0.404; P = 0.022) and between TGF-beta1 AI and carotid artery intima-media thickness (r = -0.587; P = 0.0004). A low AI was associated with irreversible damage (SLICC [Systemic Lupus International Collaborating Clinics] Damage Index > or = 1) and was inversely correlated with disease duration. Intima-media thickness was significantly linked to total cholesterol (r = 0.371; P = 0.037). To conclude, in SLE low normal TGF-beta1 activation was linked with increased lymphocyte apoptosis, irreversible organ damage, disease duration, calculated low-density lipoprotein levels and increased carotid IMT, and may contribute to the development of early atherosclerosis.

Topics: Apoptosis; Atherosclerosis; Cholesterol, LDL; England; Female; Humans; Lupus Erythematosus, Systemic; Lymphocytes; Male; Reference Values; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tunica Media; White People

Regulatory T cells have an important role in the control of self-reactivity, and in the pathogenesis of autoimmune inflammatory conditions. The aim of this work was to perform a quantitative and functional analysis of regulatory T cells in patients with systemic lupus erythematosus (SLE). We studied twenty-three patients with SLE (19 active, 4 inactive), and twenty-seven healthy subjects as well as fifteen patients with rheumatoid arthritis (RA). The following cell subsets were analyzed in peripheral blood mononuclear cells by flow cytometry: CD4+CD25+, CD4+CD25(bright), CD4+Foxp3+ (Treg cells), CD8+CD28- (Ts cells), CD4+IL-10+ (Tr1 cells), and CD4+TGF-beta+ (Th3 cells). In addition, the in vitro suppressive activity of CD4+CD25+ lymphocytes was tested. We found no significant differences in the levels of all regulatory cell subsets studied in SLE patients compared to controls and RA patients. However, a defective regulatory function of CD4+CD25+T cells was observed in a significant fraction (31%) of patients with SLE. Our data indicate that although approximately one third of patients with SLE show an abnormal immunosuppressive function of Treg lymphocytes, their levels of the different regulatory T cell subsets in peripheral blood are not significantly different from those found in controls.

Topics: Adolescent; Adult; Arthritis, Rheumatoid; Female; Flow Cytometry; Humans; Interleukin-10; Lupus Erythematosus, Systemic; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is a chronic, systemic autoimmune disease characterized by the loss of tolerance to self-antigen. Because it is currently not known if regulatory T (T(reg)) cells are involved in the pathogenesis, we determined the frequency of CD4(+)CD25(+) T cells and assayed the related gene expression levels in CD4(+)CD25(+) T cells isolated from both lupus mice (NZB/NZW F(1)) and normal control mice (DBA2/NZW F(1)). The results showed that the frequency of CD4(+)CD25(+) T cells in lupus mice was lower than that of normal mice. Except for the high expression level of interleukin (IL)-10 mRNA, CD4(+)CD25(+) T cells from lupus mice expressed normal forkhead box P3 (Foxp3) and transforming growth factor (TGF)-beta mRNA, and exerted suppressive functions. Furthermore, we depleted CD25(+) T(reg) cells of non-autoimmune mice with anti-CD25 antibody and broke their tolerance with apoptotic cell-pulsed dendritic cells for the follow-up of autoantibody levels. The mice in the CD25(+) cell-depleted group had higher titres of anti-double-strand/single-strand DNA antibodies than those of the isotype control antibody-treated group. These findings indicated that CD4(+)CD25(+) T cells might be involved in the regulatory mechanism of autoantibody production.

Topics: Animals; Apoptosis; Autoantibodies; CD4-Positive T-Lymphocytes; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Forkhead Transcription Factors; Gene Expression; Immunosuppression Therapy; Interleukin-10; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Receptors, Interleukin-2; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transforming Growth Factor beta

A peptide (hCDR1) based on the complementarity determining region-1 of an anti-DNA antibody ameliorates systemic lupus erythematosus (SLE) in induced and spontaneous lupus models. Our objectives were to determine the effects of hCDR1 on TCR signaling and on its negative regulators, Foxj1 and Foxo3a. BALB/c mice were immunized with the SLE-inducing anti-DNA antibody, designated 16/6Id, and treated with hCDR1. hCDR1 treatment specifically inhibited IFN-gamma secretion by T cells in association with down-regulated T-bet expression and NF-kappaB activation; however, GATA-3 expression was not affected. Furthermore, TCR signaling (ZAP-70 phosphorylation) was inhibited, and the mRNA expression of the two modulators of Th1 activation, Foxj1 and Foxo3a, was significantly up-regulated. The latter were also elevated in SLE-afflicted (NZBxNZW)F1 mice that were treated with hCDR1. Addition of TGF-beta, which was elevated following treatment with hCDR1, to T cells from 16/6Id immunized mice, up-regulated Foxj1 and Foxo3a mRNA expression, similarly to hCDR1. In contrast, anti-TGF-beta antibodies added to hCDR1-treated T cells abrogated its effect. Thus, hCDR1 elevates TGF-beta, which contributes to the up-regulation of T cell Foxj1 and Foxo3a expression, leading to inhibition of NF-kappaB activation and IFN-gamma secretion, which is required for the maintenance of SLE.

Topics: Animals; Antibodies, Monoclonal; Female; Forkhead Box Protein O3; Forkhead Transcription Factors; Interferon-gamma; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; NF-kappa B; Peptide Fragments; Peptides; Phosphorylation; Receptors, Antigen, T-Cell; RNA, Messenger; T-Box Domain Proteins; T-Lymphocytes; Transforming Growth Factor beta; Up-Regulation; ZAP-70 Protein-Tyrosine Kinase

Systemic lupus erythematosus (SLE) can be induced in mice by immunizing them with a monoclonal human anti-DNA Ab that expresses a major Id, designated 16/6Id. In addition, a peptide based on the sequence of the CDR 1 (hCDR1) of the 16/6Id ameliorated the clinical manifestations of SLE in experimental models. In this study we examined the effects of treating mice with human complementary-determining region 1 (hCDR1) on the subsequent chemotaxis of T cells derived from 16/6Id-primed mice. First we demonstrated elevated levels of stromal cell-derived factor-1alpha (SDF-1alpha) in the sera of SLE-afflicted mice and in the sera and lymphoid tissues of 16/6Id-immunized BALB/c mice shortly after the immunization. We then found that administration of hCDR1 to 16/6Id-immunized mice specifically down-regulated SDF1alpha-induced T cell chemotaxis through fibronectin and collagen type I. This was accompanied by diminished SDF1-alpha-induced T cell adhesion and ERK phosphorylation. Treatment with hCDR1 up-regulated TGF-beta secretion, which, in turn, inhibited the murine T cell adhesion to and chemotaxis through fibronectin as well as their ERK phosphorylation. Thus, the secretion of TGF-beta after treatment of 16/6Id-immunized mice with hCDR1 plays an important role in the down-regulation of SDF-1alpha-mediated T cell activation and the interactions with extracellular matrix moieties observed in the present study.

Topics: Animals; Antibodies, Antinuclear; Blotting, Western; Cell Adhesion; Chemokine CXCL12; Chemokines, CXC; Chemotaxis, Leukocyte; Complementarity Determining Regions; Disease Models, Animal; Down-Regulation; Fibronectins; Humans; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Peptides; T-Lymphocytes; Transforming Growth Factor beta; Up-Regulation

We induced very low-dose tolerance by injecting lupus prone (SWR x NZB)F1 (SNF1) mice with 1 mug nucleosomal histone peptide autoepitopes s.c. every 2 wk. The subnanomolar peptide therapy diminished autoantibody levels and prolonged life span by delaying nephritis, especially by reducing inflammatory cell reaction and infiltration in kidneys. H4(71-94) was the most effective autoepitope. Low-dose tolerance therapy induced CD8+, as well as CD4+ CD25+ regulatory T (Treg) cell subsets containing autoantigen-specific cells. These adaptive Treg cells suppressed IFN-gamma responses of pathogenic lupus T cells to nucleosomal epitopes at up to a 1:100 ratio and reduced autoantibody production up to 90-100% by inhibiting nucleosome-stimulated T cell help to nuclear autoantigen-specific B cells. Both CD4+ CD25+ and CD8+ Treg cells produced and required TGF-beta1 for immunosuppression, and were effective in suppressing lupus autoimmunity upon adoptive transfer in vivo. The CD4+ CD25+ T cells were partially cell contact dependent, but CD8+ T cells were contact independent. Thus, low-dose tolerance with highly conserved histone autoepitopes repairs a regulatory defect in systemic lupus erythematosus by generating long-lasting, TGF-beta-producing Treg cells, without causing allergic/anaphylactic reactions or generalized immunosuppression.

Topics: Adoptive Transfer; Amino Acid Sequence; Animals; Antibodies, Antinuclear; Autoantigens; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Epitopes; Female; Immune Tolerance; Immunosuppression Therapy; Lupus Erythematosus, Systemic; Lupus Nephritis; Mice; Mice, Inbred NZB; Molecular Sequence Data; Nuclear Proteins; Nucleosomes; Peptides; Receptors, Interleukin-2; T-Lymphocyte Subsets; Transforming Growth Factor beta

We investigated the expression profile of inflammatory cytokines in the lung of lupus-prone MRL/lpr mice and evaluated the therapeutic potential of mycophenolate mofetil (MMF) in reducing pulmonary cytokines in active lupus. Eight-week old female MRL/lpr mice (n = 20) were treated with MMF in vehicle by oral gavage. Disease control MRL/lpr mice (n = 30) or normal control MRL mice (n = 20) received vehicle alone. The mice were sacrificed after eight or 12 weeks of treatment. Gene expression and protein synthesis of IL-1beta, MCP-1 and TGF-beta1 in lung tissues were determined. We found an increase in the gene expression of IL-1beta, MCP-1 and TGF-beta1 in lung tissues of untreated MRL/lpr mice compared with MRL mice at either 16 weeks or 20 weeks of age. MMF treatment significantly prolonged the survival of MRL/lpr mice, down-regulated the gene expression of IL-1beta, MCP-1 and TGF-beta1 in lung tissues at the end of eight or 12 weeks of treatment. Protein synthesis of TGF-beta1 was decreased following eight weeks of MMF treatment. We conclude that MMF treatment can reduce the TGF-beta1 gene expression and protein synthesis in lung tissues of lupus-prone mice. Our findings provide experimental data suggesting a beneficial potential of MMF therapy in pulmonary involvement of lupus.

Topics: Animals; Chemokine CCL2; Female; Immunosuppressive Agents; Interleukin-1; Lung; Lupus Erythematosus, Systemic; Mice; Mice, Inbred MRL lpr; Mycophenolic Acid; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1

The role of CD4(+)CD25+ T cell in glomerulonephritis (GN) development during the preactive phase was investigated in autoimmune-prone female NZB x NZW F1 (B/WF1) mice. The administration of anti-mouse CD25+ T-cell monoclonal antibody (PC61.5) 3 days after birth induced the development of GN with an increase in IgG2a antinuclear antibody, productions of IL-6 and IFN-gamma, whereas TGF-beta1 production decreased, compared to untreated control mice. The present study results suggest that CD4(+)CD25+ T cells may, at least in part, downregulate the development of GN during the preactive phase in B/WF1 mice.

Topics: Acute Disease; Animals; Antibodies, Monoclonal; Autoimmune Diseases; Biomarkers; CD4 Antigens; Complement C3a; Female; Glomerulonephritis; Immunoglobulin A; Immunohistochemistry; Interferon-gamma; Interleukin-4; Interleukin-6; Kidney; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Models, Animal; Receptors, Interleukin-2; T-Lymphocytes; Transforming Growth Factor beta

Cytokine network alterations contribute strongly to the initiation and perpetuation of systemic lupus erythematosus (SLE). This study investigated the cytokines production and polymorphism association of cytokines with SLE in Taiwan. The cytokines investigated included tumor necrosis factor-alpha (TNF-alpha), TNF-beta (TNF-beta), interleukin (IL)-4, IL-10 and transforming growth factor-beta1 (TGF-beta1).. Genotyping of different cytokine genes was performed using polymerase chain reaction and restriction fragment length polymorphism methods in 136 patients. The cytokine levels in the supernatants of cultures of peripheral blood mononuclear cells (PBMCs) were determined by enzyme immunoassay.. The rates of genotype polymorphism of TNF-beta +252, IL-4 -590 and IL-10 -819 were significantly correlated with SLE. However, only the genotype frequency of TNF-beta +252A was in accordance with Hardy-Weinberg equilibrium and significantly greater than in the normal group. The stimulated PBMC culture supernatants from TNF-beta +252A carriers produced lower levels of TNF-beta compared to TNF-beta +252G/G homozygotes. Moreover, TNF-beta levels in stimulated PBMC culture supernatants were negatively correlated with those of IL-4, IL-10 and TGF-beta1. It is possible that TNF-beta plays a modulatory role in the expression of these 3 cytokines.. TNF-beta +252A polymorphism, other than TNF-alpha, IL-4, IL-10, and TGF-beta1, is significantly associated with SLE in Taiwan.

Topics: Adult; Female; Genotype; Humans; Interleukin-10; Interleukin-4; Linkage Disequilibrium; Lupus Erythematosus, Systemic; Lymphotoxin-alpha; Male; Polymorphism, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Platelets are potential contributors to glomerular injury via the release of chemotactic and/or mitogenic mediators upon activation or through direct CD154/CD40-dependent interaction with cell components of the glomerulus. We examined whether platelets could activate mesangial cells and the potential role of the platelet-associated CD154.. Thrombin-activated platelets from systemic lupus erythematosus (SLE) patients or from disease or healthy controls were grown with human mesangial cells in the presence or not of a neutralizing anti-CD154 antibody either in contact or in a noncontact setting, the platelets and mesangial cells being separated by a pore size semipermeable membrane. The induction of mesangial cell surface antigens was assayed by flow cytometry. The quantification of mesangial cell proliferation was performed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and the production of transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor (PDGF) and soluble CD40 were measured by enzyme-linked immunosorbent assay (ELISA).. Activated platelets from patients with SLE could induce an up-regulation of the expression of CD40 on mesangial cells with a concomitant release of soluble CD40. This induction required a direct contact between platelets and mesangial cells and was dependent upon the platelet-associated CD154. Pathologic consequences of the up-regulation of CD40 were a CD40-dependent stimulation of the proliferation of mesangial cells and a CD40-dependent increased production of TGF-beta1 by these cells.. Platelets from patients with SLE can activate mesangial cells through CD40/CD154 interactions, leading to an induction of proliferation of the mesangial cells and an enhanced production of TGF-beta1, a profibrotic cytokine.

Topics: Adult; Aged; Blood Platelets; CD40 Antigens; CD40 Ligand; Cell Communication; Cells, Cultured; Coculture Techniques; Female; Humans; Lupus Erythematosus, Systemic; Male; Mesangial Cells; Middle Aged; Platelet Activation; Transforming Growth Factor beta; Transforming Growth Factor beta1; Up-Regulation

Systemic lupus erythematosus (SLE), which is characterized by the increased production of autoantibodies and defective T cell responses, can be induced in mice by immunization with a human anti-DNA mAb that expresses a major Id, designated 16/6Id. A peptide based on the sequence of the CDR1 of the 16/6Id (human CDR1 (hCDR1)) ameliorated the clinical manifestations of SLE and down-regulated, ex vivo, the 16/6Id-induced T cell proliferation. In this study, we examined the mechanism responsible for the hCDR1-induced modulation of T cell functions related to the pathogenesis of SLE. We found that injection of hCDR1 into BALB/c mice concomitant with their immunization with 16/6Id resulted in a marked elevation of TGF-beta secretion 10 days later. Addition of TGF-beta suppressed the 16/6Id-stimulated T cell proliferation similarly to hCDR1. In addition, we provide evidence that one possible mechanism underlying the hCDR1- and TGFbeta-induced inhibition of T cell proliferation is by down-regulating the expression, and therefore the functions, of a pair of key cell adhesion receptors, LFA-1 (alphaLbeta2) and CD44, which operate as accessory molecules in mediating APC-T cell interactions. Indeed, T cells of mice treated with hCDR1 showed a TGF-beta-induced suppression of adhesion to the LFA-1 and CD44 ligands, hyaluronic acid and ICAM-1, respectively, induced by stromal cell-derived factor-1alpha and PMA. The latter suppression is through the inhibition of ERK phosphorylation. Thus, the down-regulation of SLE-associated responses by hCDR1 treatment may be due to the effect of the up-regulated TGF-beta on the expression and function of T cell adhesion receptors and, consequently, on T cell stimulation, adhesion, and proliferation.

Topics: Animals; Antibodies, Antinuclear; Antibodies, Monoclonal; Autoimmunity; Cell Adhesion; Cell Communication; Cell Proliferation; Complementarity Determining Regions; Disease Models, Animal; Flow Cytometry; Humans; Hyaluronan Receptors; Hyaluronic Acid; Immunoglobulin Idiotypes; Intercellular Adhesion Molecule-1; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Function-Associated Antigen-1; Mice; Mice, Inbred BALB C; Peptides; T-Lymphocytes; Transforming Growth Factor beta

Lupus-prone (NZB x NZW)F1 mice spontaneously develop elevated titers of anti-DNA Abs that contain T cell determinants in their V(H) regions. We have previously shown that tolerization with an artificial peptide based on these T cell determinants (pConsensus (pCons)) can block production of anti-DNA Abs and prolong survival of the mice. In this study, we show that this protection depends in part on the generation of peripheral TGFbeta- and Foxp3-expressing inhibitory CD8+ (Ti) cells. These CD8+ Ti cells suppress anti-DNA IgG production both in vitro and in vivo and require up-regulated expression of both Foxp3 and TGFbeta to exert their suppressive function, as indicated by microarray analyses, small interfering RNA inhibition studies, and blocking experiments. Additionally, CD8+ Ti cells from pCons-tolerized mice were longer-lived suppressors that up-regulated expression of Bcl-2 and were more resistant to apoptosis than similar cells from naive mice. These data indicate that clinical suppression of autoimmunity after administration of pCons depends in part on the generation of CD8+ Ti cells that suppress secretion of anti-DNA Ig using mechanisms that include Foxp3, TGFbeta, and resistance to apoptosis.

Topics: Adoptive Transfer; Animals; Antibodies, Antinuclear; Apoptosis; CD8-Positive T-Lymphocytes; Female; Flow Cytometry; Forkhead Transcription Factors; Immune Tolerance; Immunoglobulin Variable Region; Lupus Erythematosus, Systemic; Mice; Mice, Inbred NZB; Microarray Analysis; Oligopeptides; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta

Regulatory T cells generated ex vivo from conventional mouse T cells have been used to prevent and alter the course of a stimulatory graft-vs-host disease with a lupus-like syndrome. DBA/2 mouse T cells induce this syndrome when injected into (DBA/2 x C57BL/6) F(1) mice. Stimulating DBA/2 T cells with irradiated C57BL/6 in the presence of IL-2 and TGF-beta induced both CD4(+) and CD8(+) cells to develop potent suppressive activity and enhanced their survival. The IL-2 and TGF-beta-treated T cells lost their ability to induce graft-vs-host disease and, instead, prevented other parental T cells from inducing lymphoid hyperplasia, B cell activation, and an immune complex glomerulonephritis. Moreover, a single transfer of TGF-beta-conditioned T cells to animals that had already developed anti-dsDNA Abs decreased the titer, suppressed proteinuria, and doubled survival. This study raises the possibility that autologous regulatory T cells generated ex vivo have the potential to be used as an adoptive immunotherapy to induce allograft tolerance and to control autoimmunity.

Topics: Adoptive Transfer; Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Differentiation; Cell Survival; Cells, Cultured; Female; Graft vs Host Disease; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred DBA; Survival Analysis; Syndrome; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Transforming growth factor-beta1 (TGF-beta1) is involved in the generation of CD8+ T suppressor cells, natural killer (NK) cells and regulatory T (Th3) cells for down-regulatory effects on antibody production. We studied TGF-beta1 activity in patients with systemic lupus erythematosus (SLE) to try to clarify whether the dysregulation by TGF-beta1 is genetically determined. Sera from 55 patients with clinically inactive SLE, who were taking minimal steroids and/or hydroxychloroquine, and 40 healthy controls, along with supernatants from concanavalin A-stimulated peripheral blood mononuclear cell (PBMC) cultures from 18 patients with SLE and 10 controls were subjected to TGF-beta1 enzyme-linked immunosorbent assay. A total of 138 patients with SLE and 182 controls were genotyped for 5 single-nucleotide polymorphisms (SNPs) of TGF-beta1: -988C/A, -800G/A, -509C/T, Leu10/Pro10 and Arg25/Pro25. Patients with SLE had lower serum levels of TGF-beta1 compared with controls (p=0.052). The unstimulated and stimulated TGF-beta1 production of PBMCs in patients with SLE was higher than in controls, although these differences did not reach significance (p=0.073 and 0.074, respectively). None of the TGF-beta1 SNPs was strongly associated with SLE in Taiwanese patients or had any prognostic significance in lupus nephritis. Hence these polymorphisms do not represent a genetic predisposition to SLE. The intrinsic capability of immunoregulation for spontaneous B cell hyperactivity through PBMC TGF-beta1 production was presumed to be intact in clinically stable SLE in Taiwanese. Whether the lower serum TGF-beta1 level that causes the defective immune regulation in SLE is primarily under genetic control or secondary to the influence of ongoing cellular interactions in the cytokine context needs to be elucidated.

Topics: Adult; Case-Control Studies; Female; Genetic Predisposition to Disease; Genotype; Humans; Lupus Erythematosus, Systemic; Lupus Nephritis; Male; Polymorphism, Single Nucleotide; Taiwan; Transforming Growth Factor beta; Transforming Growth Factor beta1

Immunization with portions of a murine antibody to DNA induced Ig peptide-reactive peripheral CD8+ inhibitory T (Ti) cells in non-autoimmune (BALB/c x NZW) F1 (CWF1) mice. Those Ti suppressed in vitro production of IgG anti-DNA by lymphocytes from MHC-matched, lupus-prone (NZB x NZW) F1 (BWF1) mice, primarily via secretion of transforming growth factor-beta (TGF-beta). However, splenic CD8+ cells from immunized BWF1 mice failed to suppress anti-DNA. Therefore, BWF1 mice were studied for defects in peripheral CD8+ T cells. The potential to suppress autoimmunity mediated by activated CD4+ helper T and B cells in BWF1 mice was assessed. As BWF1 mice aged, peripheral CD8+ T cells expanded little; fewer than 10% displayed surface markers of activation and memory. In contrast, quantities of splenic CD4+ T and B cells increased; high proportions displayed activation/memory markers. In old compared to young BWF1 mice, splenic cell secretion of two cytokines required for generation of CD8+ T effectors, IL-2 and TGF-beta, was decreased. Immunizing BWF1 mice activated peptide-reactive CD8+ T cells, but their number was decreased compared to young BWF1 or old normal mice. While peptide-reactive splenic CD8+ T cells from immunized BWF1 mice did not survive in short-term cultures, similar CD8+ T cell lines from immunized CWF1 mice expanded and on transfer into BWF1 mice delayed autoimmunity and prolonged survival. Therefore, CD8+ T cells in old BWF1 mice are impaired in expansion, acquisition of memory, secretion of cytokine, and suppression of autoimmunity. Understanding these defects might identify targets for therapy in systemic lupus erythematosus.

Topics: Animals; Apoptosis; Autoantibodies; Autoimmunity; CD8-Positive T-Lymphocytes; Disease Models, Animal; Female; Immunization; Immunologic Memory; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Inbred NZB; Transforming Growth Factor beta; Transforming Growth Factor beta1

Two peptides, based on the sequences of the complementarity-determining regions (CDR) 1 and 3 of a pathogenic murine monoclonal anti-DNA autoatibody that bears the 16/6 idiotype (Id), were shown to either prevent or treat an already established systemic lupus erythematosus (SLE) in two murine models of lupus. Two additional peptides based on the human monoclonal anti-DNA, 16/6 Id were synthesized. This study was undertaken in order to investigate the ability of the CDR-based peptides to immunomodulate SLE-associated responses of peripheral blood lymphocytes (PBL) of SLE patients. PBL of 24 of the 62 SLE patients tested proliferated in vitro following stimulation with the human 16/6 Id. Peptides based on the CDRs of both the human and murine anti-DNA autoantibodies inhibited efficiently and specifically the 16/6 Id-induced proliferation and IL-2 production. The latter inhibitions correlated with an up-regulated production (by 2.5-3.5-fold) of the immunosuppressive cytokine, TGF-beta. Overall, the results of our study demonstrate that the CDR-based peptides are capable of down-regulating in vitro autoreactive T cell responses of PBL of SLE patients. Thus, these peptides are potential candidates for a novel specific treatment of SLE patients.

Topics: Adolescent; Adult; Amino Acid Sequence; Animals; Antibodies, Antinuclear; Cell Division; Cells, Cultured; Child; Complementarity Determining Regions; Female; Humans; Immune Tolerance; Interleukin-2; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Mice; Middle Aged; Molecular Sequence Data; Peptide Fragments; Transforming Growth Factor beta; Up-Regulation

Lymphocyte production of transforming growth factor (TGF)-beta1 is decreased in systemic lupus erythematosus (SLE). The lack of this immunoregulatory cytokine may contribute to the characteristic T cell disregulation and aberrant B cell stimulation in SLE patients. The less common C allele of the TGFB1 polymorphism (G915C) is associated with a lower TGF-beta1 production capacity. We performed a population-based case-control study to analyse the impact of this polymorphism on disease susceptibility, on clinical SLE manifestations and autoantibody production. A total of 203 German Caucasian SLE patients (fulfilling the 1982 ACR disease duration 11.5 +/- 7.0 years) and 158 ethnically, age- and sex-matched healthy controls were genotyped with a mutagenically separated polymerase chain reaction. There were no significant differences in the genotype distribution and allele frequencies between patients (915 C = 0.08) and healthy controls (915 C = 0.10). Comparing subgroups of patients, we found no association of major disease manifestations or specific autoantibodies with TGFB1 genotypes or alleles. The TGFB1 polymorphism (G915C) neither significantly contributes to the disease susceptibility, nor predisposes to clinical and immunological manifestations typical of SLE. Further studies are needed to corroborate the pathogenic role of TGF-beta1 in SLE patients and to identify the precise genetic elements controlling its production.

Topics: Adult; Case-Control Studies; Female; Genetic Predisposition to Disease; Humans; Lupus Erythematosus, Systemic; Lymphocytes; Male; Polymorphism, Genetic; Transforming Growth Factor beta; Transforming Growth Factor beta1

In contrast to necrotic cells, the clearance of apoptotic ones usually is an anti-inflammatory process which elicits only a marginal immune response. During apoptosis phosphatidylserine (PS) is exposed on the outer leaflet of the cytoplasmic membrane and serves as target for the PS receptor of phagocytes. The latter is responsible for anti-inflammatory signalling and the induction of TGFbeta. We were interested whether the immunogenicity of apoptotic cells can be increased by masking PS. We observed that treatment of xenogeneic apoptotic cells with annexin V (AxV) significantly increased the humoral immune response against surface epitopes of these cells. Furthermore, AxV-coated irradiated tumour cells were able to elicit a long lasting tumour specific cytotoxic T lymphocyte response. AxV efficiently blocked the uptake of irradiated cells by macrophages but not by dendritic cells. Furthermore, AxV skewed the phagocytosis of irradiated cells towards inflammation. Investigation of patients with autoimmune diseases further supported the role of anionic surface phospholipids for anti-inflammatory clearance of apoptotic cells. Impaired clearance and opsonisation with anti-phospholipid-antibodies are discussed to be responsible for the development of systemic lupus erythematosus and anti-phospholipid-syndrome, respectively. Presentation of cryptic epitopes from late apoptotic cells in a proinflammatory context may challenge T cell tolerance. In addition, accumulation of uncleared apoptotic debris in the germinal centres of lymph nodes may result in the survival of autoreactive B cells.

Topics: Animals; Anions; Annexin A5; Anti-Inflammatory Agents; Antiphospholipid Syndrome; Apoptosis; Cell Line; Cells, Cultured; Chickens; Cytoplasm; Dendritic Cells; Female; Flow Cytometry; Humans; Immunohistochemistry; Immunosuppressive Agents; In Situ Nick-End Labeling; Inflammation; Lupus Erythematosus, Systemic; Lymph Nodes; Macrophages; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence; Phagocytes; Phagocytosis; Phosphatidylserines; Phospholipids; Protein Binding; Signal Transduction; T-Lymphocytes; Transforming Growth Factor beta; Tumor Cells, Cultured

We studied a well-selected population of patients with active rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) without immunosuppressive therapy. Control and patient peripheral blood mononuclear cells (PBMC) were incubated with IL-1beta, IL-10, TGF-beta or LPS for 20 h and the in vitro basal and stimulated secretions of IL-6, TNF-alpha, IL-1beta and IL-1ra were measured by ELISA. We found that in the SLE patients the basal secretion of IL-6 was significantly lower and that of IL-1ra significantly higher than in control subjects, while in the RA group the basal IL-1ra secretion was higher than in healthy subjects. SLE and RA PBMC responded to LPS and IL-1beta reaching higher cytokine secretion values than controls. The in vitro response of SLE and RA PBMC to TGFbeta was normal, while that to IL-10 was defective: IL-10 was able to stimulate the production of IL-6 and IL-1ra in PBMC from normal subjects, but it was unable to enhance IL-6 secretion in RA cells and it was also completely ineffective in inducing IL-1ra secretion in both SLE and RA PBMC. Our work add new data useful for the evaluation of IL-10 and IL-1ra as therapeutic agents in rheumatic diseases.

Topics: Adolescent; Adult; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Case-Control Studies; Cytokines; Female; Humans; In Vitro Techniques; Inflammation Mediators; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-10; Interleukin-6; Leukocytes, Mononuclear; Lipopolysaccharides; Lupus Erythematosus, Systemic; Middle Aged; Recombinant Proteins; Sialoglycoproteins; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To determine if fatigue in patients with systemic lupus erythematosus (SLE) is associated with levels of serum cytokines, antiphospholipid antibodies (aPL), or other disease features.. In a cross sectional study 57 Caucasian patients with SLE were subjected to clinical neurological examination and cerebral magnetic resonance imaging (MRI). Fatigue was evaluated by Fatigue Severity Scale (FSS) and disease activity by SLE Disease Activity Index (SLEDAI). Serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 2 (IL-2), IL-6, IL-10, transforming growth factor-beta (TGF-beta), interferon-alpha (IFN-alpha), anticardiolipin antibody (aCL) IgG and IgM, as well as anti-beta2-glycoprotein I antibody (anti-beta2-GPI) IgG and IgM were analyzed by ELISA.. Four of 5 patients with SLE had fatigue (FSS score > or = 3). There were no associations between fatigue and any sociodemographic variables, medication for SLE, disease activity, cerebral infarcts, serum cytokines, aCL or beta2-GPI antibodies, or any routine hematological, biochemical, or immunological tests.. Fatigue is a common phenomenon in patients with SLE. There is no association to disease activity or other markers of disease or inflammation. Fatigue is a complex phenomenon, and cytokine involvement in brain tissue not reflected by cytokine serum concentrations in this study cannot be excluded. Alternatively, psychosocial factors may well be the dominant predictor of fatigue in patients with SLE.

Topics: Adult; Aged; Antibodies, Antiphospholipid; Cross-Sectional Studies; Cytokines; Fatigue; Female; Humans; Interferon-alpha; Interleukin-10; Interleukin-2; Interleukin-6; Lupus Erythematosus, Systemic; Male; Middle Aged; Severity of Illness Index; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To determine whether transforming growth factor-beta2 (TGF-beta2) gene polymorphism is associated with systemic lupus erythematosus (SLE) susceptibility. TGF-beta is a multifunctional family of cytokines important in tissue repair, inflammation and immunoregulation. SLE is thought to be a T cell dependent autoimmune disorder with T cell dysfunction. Due to its known suppressive effects on interleukin 2 dependent T cell growth, TGF-beta2 is considered to be a candidate SLE susceptibility gene. Furthermore, SLE has been linked with a region to which the TGF-beta2 gene has been mapped.. Association studies were performed in 3 case-control populations, from Spain. Turkey, and UK, using a TGF-beta2 5'-untranslated region (5'-UTR) 4 base pair (bp) insertion polymorphism. Genotyping was performed using fluorescent labeled polymerase chain reaction product sizing. Results. No significant differences were detected in TGF-beta2 5'-UTR polymorphism allele frequencies between SLE patients and matched controls in the 3 populations studied.. The 4 bp insertion polymorphism within the TGF-beta2 gene does not appear to be associated with SLE. However, this does not rule out the possible involvement of TGF-beta2 in the disease pathogenesis.

Topics: Alleles; Base Sequence; Case-Control Studies; Chi-Square Distribution; Cohort Studies; Female; Gene Frequency; Genetic Predisposition to Disease; Humans; Lupus Erythematosus, Systemic; Male; Molecular Sequence Data; Polymerase Chain Reaction; Polymorphism, Genetic; Probability; Reference Values; Sensitivity and Specificity; Spain; Transforming Growth Factor beta; Transforming Growth Factor beta2; Turkey; United Kingdom

Many individuals develop a single or a few brief episodes of autoimmunity from which they recover. Mechanisms that quell pathologic autoimmunity following such a breakdown of self-tolerance are not clearly understood. In this study, we show that in nonautoimmune mice, dsDNA-specific autoreactive B cells exist but remain inactive. This state of inactivation in dsDNA-specific B cells could be disrupted by autoreactive Th cells; in this case T cells that react with peptides from the V(H) region of anti-DNA Abs (hereafter called anti-V(H) T cells). Immunization with anti-DNA mAb, its gamma-chain or peptides derived from its V(H) region induced anti-V(H) Th cells, IgG anti-dsDNA Ab, and proteinuria. The breakdown of B cell tolerance in nonautoimmune mice, however, was short-lived: anti-DNA Ab and nephritis subsided despite subsequent immunizations. The recovery from autoimmunity temporally correlated with the appearance of T cells that inhibited anti-DNA Ab production. Such inhibitory T cells secreted TGFbeta; the inhibition of anti-DNA Ab production by these cells was partly abolished by anti-TGFbeta Ab. Even without immunization, nonautoimmune mice possess T cells that can inhibit autoantibody production. Thus, inhibitory T cells in nonautoimmune mice may normally inhibit T-dependent activation of autoreactive B cells and/or reverse such activation following stimulation by Th cells. The induction of such inhibitory T cells may play a role in protecting nonautoimmune mice from developing chronic autoimmunity.

Topics: Amino Acid Sequence; Animals; Antibodies, Antinuclear; Antigen-Antibody Reactions; Autoantibodies; Autoantigens; Autoimmune Diseases; B-Lymphocyte Subsets; Cell Line; Clonal Anergy; Crosses, Genetic; DNA; Female; Histocompatibility Antigens Class II; Hybridomas; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred NZB; Molecular Sequence Data; Proteinuria; Self Tolerance; Species Specificity; Spleen; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Cutaneous lupus erythematosus (LE) often presents clinically as chronic cutaneous lesions, healing with scar formation, and acute cutaneous lesions that are seen in systemic and subacute LE and heal without scarring. UV-light plays a role in the pathogenesis of the skin lesions, but the pathomechanism is still unclear. The aim of this study was to compare fibroblast proliferation and response to UV-light by cultured fibroblasts from scarring and non-scarring LE lesions. Fibroblasts were cultured from skin lesions from 5 patients with classic discoid LE, 5 patients with subacute cutaneous LE and healthy, age-matched donors. Proliferation rate was assessed by cell counts at days 3, 6 and 9. The fibroblast cultures were irradiated with UVA and the supernatants were analysed for IL-6, TGF-beta, IL-4, soluble ICAM-1 and soluble VCAM-1. Fibroblast cultures from scarring lesions showed significantly lower cell-counts at days 3 (P = 0.01) and 9 (P = 0.009), than cultures from nonscarring lesions or controls. There were no significant differences in levels of IL-6 or TGF-beta in supernatants of irradiated fibroblasts from patients compared to controls and IL-4 and the soluble forms of ICAM-1 and VCAM-1 were below detection level. The response to UV-irradiation was similar to that of normal cells in the parameters studied. In summary, cultured fibroblasts from scarring LE lesions displayed significantly decreased proliferation rates compared to non-scarring LE lesions and controls. This may be secondary to inflammatory factors, or due to a functional defect in LE fibroblasts.

Topics: Cell Division; Cells, Cultured; Fibroblasts; Humans; Intercellular Adhesion Molecule-1; Interleukin-4; Interleukin-6; Lupus Erythematosus, Systemic; Transforming Growth Factor beta; Ultraviolet Rays; Vascular Cell Adhesion Molecule-1

We have investigated the effects of interleukin-2 (IL-2) and transforming growth factor-beta (TGF-beta) gene therapy on the progress of autoimmune disease in MRL-lpr/lpr mice, a murine model of systemic lupus erythematosus (SLE). These mice have uncontrolled proliferation of T cells, an impaired response to T cell mitogen and produce autoantibodies against nuclear antigens, including DNA. Immune complexes formed by these autoantibodies are believed to cause glomerulonephritis and vasculitis in lupus mice and human SLE. Since there is an imbalance of cytokine production in both SLE patients and lupus mice, we examined the effects of cytokine gene therapy on the progression of autoimmune disease in MRL-lpr/lpr mice. The mice were treated orally with a non-pathogenic strain of Salmonella typhimurium bearing the aroA-aroD- mutations and carrying the murine genes encoding IL-2 and TGF-beta. The bacteria synthesise and slowly release the cytokines in vivo. Our results show that, contrary to expectation, TGF-beta gene therapy produced no improvement in pathology and generally had opposite effects to those of IL-2. IL-2 gene therapy restored the defective T cell proliferative response to mitogen and suppressed the autoantibody response, glomerulonephritis and growth of lymphoid tumours.

Topics: Animals; Antibodies, Antinuclear; Female; Genetic Therapy; Interleukin-2; Kidney; Lupus Erythematosus, Systemic; Lymphocyte Activation; Mice; Mice, Inbred MRL lpr; Salmonella typhimurium; T-Lymphocytes; Transforming Growth Factor beta

To define the phenotype of cells in the perivascular and vascular infiltrates of Palmerston North (PN) mice and the cytokines that those cells produce.. Immunohistologic analysis, flow cytometric analysis, and reverse transcriptase-polymerase chain reaction (RT-PCR) studies were performed on tissues and cells from female PN mice and age-matched and sex-matched DBA/2 mice.. With aging, PN mice developed a female-predominant, lupus-like disease, with a severe systemic mononuclear cell perivasculitis and vasculitis. The perivasculitis involved arteries and veins in kidney, liver, brain, and lung; the vasculitis predominantly involved veins and venules. The perivascular and vascular infiltrates in female PN mice were composed mainly of an unusual cell type that expressed phenotypic markers characteristic of both T cells (Thy1+, CD3+, CD4+, T cell receptor + [TCR+]) and B cells (B220+). In addition, the infiltrates contained a smaller number of conventional CD4+,B220- T cells and macrophages. Very few CD8+ T cells or surface Ig+ B cells were seen. Unlike the Thy1+,B220+ T cells present in MRL/lpr mice, most of which were CD4-,CD8- and TCRalpha/beta+, the majority of the Thy1+,B220+ T cells in the perivascular/vascular infiltrates of PN mice were CD4+ and expressed either TCRalpha/beta or TCRgamma/delta. By immunohistologic staining, the cells in the perivascular and vascular infiltrates in the kidneys of older PN mice were shown to produce interleukin-4 (IL-4), IL-6, and IL-10, but not IL-2, interferon-gamma, transforming growth factor beta, tumor necrosis factor alpha, or IL-1beta. By RT-PCR, the kidneys of older PN mice were found to express high levels of IL-4, IL-6, and IL-10 messenger RNA.. The vascular and perivascular infiltrates in PN mice are composed predominantly of an unusual subpopulation of T cells that are Thy1+,B220+,CD4+,CD8-, express either TCRalpha/beta or TCRgamma/delta, and produce mainly type 2 cytokines. The exact role of these cells in the immunopathogenesis of lupus-like disease in PN mice remains to be elucidated.

Topics: Animals; B-Lymphocytes; Disease Models, Animal; Female; Gene Expression; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-2; Interleukin-4; Interleukin-6; Lupus Erythematosus, Systemic; Mice; Mice, Inbred DBA; Mice, Inbred MRL lpr; Mice, Inbred NZB; Mice, Inbred Strains; Phenotype; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; T-Lymphocytes; Transforming Growth Factor beta; Vasculitis

Topics: Humans; Lupus Erythematosus, Systemic; Transforming Growth Factor beta

Transforming growth factor beta (TGF-beta) comprises of a family of proteins with pleiotropic signaling properties and potent immunoregulatory effects. In SLE we recently reported that lymphocyte production of the total and active forms of TGF-beta1 was decreased. Here we asked whether these defects correlate with disease activity and/or severity. TGF-beta1 production by blood lymphocytes from 17 prospectively studied SLE patients was compared with 10 rheumatoid arthritis (RA) patients and 23 matched healthy controls. The RA levels of active TGF-beta1 were lower than controls, but were not deceased to the extent found in SLE. Levels of constitutive and anti-CD2 stimulated active TGF-beta1 detected in picomolar amounts were markedly reduced in six untreated patients hospitalized with recent onset, very active and severe SLE and similarly reduced in 11 patients with treated, less active disease. By contrast, decreased production of total TGF-beta1 inversely correlated with disease activity. These studies suggest, therefore, that although impaired lymphocyte secretion of the latent precursor of TGF-beta1 may result as a consequence of disease activity, a decreased capacity to convert the precursor molecule to its active form may pre-date disease onset. Insufficient exposure of T cells to picomolar concentrations of TGF-beta1 at the time they are activated can result in impaired down-regulation of Ig synthesis. Therefore, decreased lymphocyte production of active TGF-beta1 in SLE could be an immunologic defect which contributes to the B cell hyperactivity characteristic of this disease.

Topics: Adult; Aged; Arthritis, Rheumatoid; CD2 Antigens; Cells, Cultured; Female; Humans; Lupus Erythematosus, Systemic; Lymphocytes; Male; Middle Aged; Severity of Illness Index; Transforming Growth Factor beta

We have recently reported that transforming growth factor-beta (TGF-beta) co-stimulates interleukin-2 (IL-2) activated CD8+ T cells to down-regulate antibody production. In SLE, lymphocyte production of both IL-2 and TGF-beta is decreased. Here we report that a brief treatment of PBMC from SLE patients with IL-2 and TGF-beta can result in marked inhibition of spontaneous polyclonal IgG and autoantibody production. Peripheral blood mononuclear cells (PBMC) from 12 patients with active SLE were exposed to IL-2 with or without TGF-beta for three days, washed and cultured for seven more days. The mean decrease in IgG secretion was 79%. The strongest inhibitory effect was observed in cases with the most marked B cell hyperactivity. Spontaneous production of anti-nucleoprotein (NP) antibodies was observed in four cases and cytokine treatment of PBMC decreased autoantibody production by 50-96%. IL-2 inhibited Ig production by either TGF-beta-dependent or independent mechanisms in individual patients. In a study of anti-CD2 stimulated IgG production in a patient with active SLE, we documented that IL-2 and TGF-beta reversed the enhancing effects of CD8+ T cells on IgG production and induced suppressive activity instead. These results raise the possibility that cytokine-mediated down-regulation of B cell hyperactivity in SLE may have therapeutic potential.

Topics: Adolescent; Adult; Antibodies, Monoclonal; B-Lymphocytes; CD2 Antigens; CD8-Positive T-Lymphocytes; Cells, Cultured; Female; Humans; Immunoglobulin G; Interleukin-2; Killer Cells, Natural; Lupus Erythematosus, Systemic; Middle Aged; Transforming Growth Factor beta

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by various autoantibodies that recognize autoantigens displayed on the surface of cells undergoing apoptosis. The genetic contribution to SLE susceptibility has been widely recognized. We previously reported evidence for linkage to SLE of the human chromosome 1q41-q42 region and have now narrowed it from 15 to 5 cM in an extended sample using multipoint linkage analysis. Candidate genes within this region include (a) PARP, poly(ADP-ribose) polymerase, encoding a zinc-finger DNA-binding protein that is involved in DNA repair and apoptosis; (b) TGFB2, encoding a transforming growth factor that regulates cellular interactions and responses; and (c) HLX1, encoding a homeobox protein that may regulate T-cell development. Using a multiallelic, transmission-disequilibrium test (TDT), we found overall skewing of transmission of PARP alleles to affected offspring in 124 families (P = 0.00008), preferential transmission of a PARP allele to affected offspring (P = 0.0003), and lack of transmission to unaffected offspring (P = 0.004). Similar TDT analyses of TGFB2 and HLX1 polymorphisms yielded no evidence for association with SLE. These results suggest that PARP may be (or is close to) the susceptibility gene within the chromosome 1q41-q42 region linked to SLE.

Topics: Alleles; Chromosome Mapping; Genetic Linkage; Humans; Linkage Disequilibrium; Lupus Erythematosus, Systemic; Poly(ADP-ribose) Polymerases; Transforming Growth Factor beta

TGF-beta has marked inhibitory effects on the immune system but also serves as a costimulatory factor in the development of T cells with down-regulatory activities. This cytokine is secreted as a latent complex and converted extracellularly to its active form. We have recently learned that anti-CD2 is a potent inducer of lymphocyte-derived TGF-beta and that NK cells are the predominant source. The objective of this study was to compare levels of constitutive, anti-CD2-induced and cytokine-regulated TGF-beta produced by blood lymphocytes from patients with systemic lupus erythematosus (SLE) in comparison with healthy controls. Using a highly sensitive and specific bioassay to assess TGF-beta, we report that unstimulated PBL from SLE patients, especially the NK cell subset, produced decreased levels of active TGF-beta. In response to anti-CD2, concentrations of active and total TGF-beta were also decreased in SLE. After learning that IL-2 and TNF-alpha enhance lymphocyte production of active TGF-beta, we found that the addition of these cytokines was unable to increase active TGF-beta to normal concentrations. Although we observed that IL-10 inhibited the production of active TGF-beta, antagonism of this cytokine was unable to completely correct the defect. In two SLE patients with B cell hyperactivity, spontaneous IgG production was almost abolished by the combination of TGF-beta and IL-2. Therefore, decreased production of each of these cytokines in SLE could be important in the perpetuation of B cell hyperactivity.

Topics: Adult; Aged; B-Lymphocytes; Cells, Cultured; Down-Regulation; Female; Humans; Immunoglobulin G; Interleukin-10; Interleukin-2; Killer Cells, Natural; Lupus Erythematosus, Systemic; Lymphocytes; Male; Middle Aged; Transforming Growth Factor beta

The level of bioactive transforming growth factor-beta (TGF-beta) was measured in serum from patients with chronic fatigue syndrome (CFS), healthy control subjects, and patients with major depression, systemic lupus erythematosis (SLE), and multiple sclerosis (MS) of both the relapsing/remitting (R/R) and the chronic progressive (CP) types. Patients with CFS had significantly higher levels of bioactive TGF-beta levels compared to the healthy control major depression, SLE, R/R MS, and CP MS groups (P < 0.01). Additionally, no significant differences were found between the healthy control subjects and any of the disease comparison groups. The current finding that TGF-beta is significantly elevated among patients with CFS supports the findings of two previous studies examining smaller numbers of CFS patients. In conclusion, TGF-beta levels were significantly higher in CFS patients compared to patients with various diseases known to be associated with immunologic abnormalities and/or pathologic fatigue. These findings raise interesting questions about the possible role of TGF-beta in the pathogenesis of CFS.

Topics: Depression; Factor Analysis, Statistical; Fatigue Syndrome, Chronic; Humans; Lupus Erythematosus, Systemic; Multiple Sclerosis; Transforming Growth Factor beta

Topics: Administration, Oral; Animals; Autoimmunity; Genetic Therapy; Genetic Vectors; Interleukin-2; Lupus Erythematosus, Systemic; Mice; Mice, Inbred MRL lpr; Salmonella typhimurium; T-Lymphocytes; Transfection; Transforming Growth Factor beta

Our study aims to determine whether anti-dsDNA exerts any effect on the gene expression of IL-8 or TGF-beta in cultured HUVEC. Both cytokines have angiogenic effect on endothelial cells. IgG was purified from 19 patients with SLE and from 19 healthy controls. Anti-dsDNA-depleted polyclonal IgG was also prepared from serum IgG of lupus patients by affinity chromatography with DNA cellulose column. Compared with either control IgG or anti-dsDNA-dep-IgG, HUVEC incubated with anti-dsDNA-containing-IgG expressed higher levels of IL-8 mRNA (p = 0.0001) and TGF-beta 1 mRNA (p = 0.0014). We demonstrated a significant increase in the percentage of cells with fragmented DNA in HUVEC incubated with anti-dsDNA-containing-IgG compared with those incubated with anti-dsDNA-dep-IgG, supporting the notion that anti-dsDNA may exert a direct apoptotic effect on cultured endothelial cells. Our study provides in vitro evidence that anti-dsDNA could play an important pathogenetic role in inducing inflammatory injury of vascular endothelium in SLE.

Topics: Adult; Antibodies, Antinuclear; Apoptosis; Cells, Cultured; DNA; DNA Fragmentation; Endothelium, Vascular; Female; Flow Cytometry; Gene Expression Regulation; Humans; Immunoglobulin G; Interleukin-8; Lupus Erythematosus, Systemic; Male; Nitric Oxide Synthase; RNA, Messenger; Transforming Growth Factor beta; Umbilical Veins

Infection remains a leading cause of morbidity and mortality in patients with SLE. To investigate this, previously we assessed the host defense status of autoimmune MRL/lpr mice and found that elaboration of active TGFbeta suppressed neutrophil function and decreased survival in response to Staphylococcus aureus infection. The purpose of the present work was to elucidate the molecular form and the cellular source of the active TGFbeta involved. Here, we report for the first time that TGFbeta1 is found in the active form inside B cells and plasma cells and that it circulates in the plasma complexed with IgG in two murine models of systemic autoimmunity and in some patients with SLE. IgG-bound active TGFbeta1 is many times more potent than uncomplexed active TGFbeta1 for suppression of neutrophil function in vitro and host defense against S. aureus infection in vivo. These data indicate that TGFbeta1 is in the active form inside B cells and plasma cells, that the formation of a complex of IgG and active TGFbeta1 is greatly accelerated in autoimmunity, and that this complex is extremely potent for suppression of PMN function and host defense against bacterial infection.

Topics: Animals; Autoimmunity; B-Lymphocytes; Humans; Immunoglobulin G; Lupus Erythematosus, Systemic; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Neutrophils; Phagocytosis; Plasma Cells; Spleen; Staphylococcal Infections; Staphylococcus aureus; Transforming Growth Factor beta

Cytokines are important in developmental and effector pathways of lymphocyte function. Our objective was to elucidate the profile of cytokines produced by circulating mononuclear cells from patients with systemic lupus erythematosus as estimated from studies of cytokine-gene activation. cDNA prepared by reverse transcription of lymphocyte mRNA was amplified using the polymerase chain reaction and normalized on the basis of beta-actin gene expression. Of 10 cytokines investigated in 16 individuals, differences between SLE and controls were found in only three. IL-2 transcripts were detected in four of six cases of subjects hospitalized for active SLE, but in only one of seven healthy controls, and none of three cases with pulmonary tuberculosis. By contrast, IL-4 transcripts were decreased compared with healthy controls and patients with tuberculosis. Also, TGF beta transcripts appeared to be decreased in SLE. All individuals studied regularly demonstrated high levels of transcripts for IL-1 beta, IL-6 and TNF alpha and transcripts for IFN gamma, TNF beta, IL-5 and IL-10 were variably expressed. In a second group of six SLE patients with less active disease, there was also a decrease in IL-4 expression compared with six healthy controls. Moreover, assays performed on sera from patients with active SLE revealed that IL-4 levels were not increased. Although in mice this cytokine has a well documented role in supporting antibody production, this study provides no evidence that IL-4 is involved in the B cell hyperactivity characteristic of human SLE.

Topics: Cytokines; Densitometry; Female; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-2; Interleukin-4; Interleukin-5; Interleukin-6; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Lymphotoxin-alpha; Male; Polymerase Chain Reaction; RNA, Messenger; T-Lymphocyte Subsets; Transcription, Genetic; Transcriptional Activation; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The urinary transforming growth factor beta (TGF-beta) excretion was measured in 33 patients including 10 with systemic lupus erythematosus (SLE), 8 with focal glomerular sclerosis (FGS), 9 with IgA nephropathy (IgAN), and 6 with membranous nephropathy (MN), and in 7 healthy subjects by enzyme-linked immunosorbent assay using a monoclonal antibody specific for TGF-beta 1 + 2 + 3. A significantly increased urinary TGF-beta excretion was observed in FGS patients (555.5 +/- 458.4 ng/mg Cr) as compared with normal controls (46.9 +/- 43.9 ng/mg Cr) (p < 0.05) and a relative increase in SLE patients (96.4 +/- 58.2 ng/mg Cr) and a decrease in MN patients (24.8 +/- 13.3 ng/mg Cr). In contrast, there was no difference in TGF-beta excretion between IgAN patients (54.1 +/- 37.4 ng/mg Cr) and normal controls. A correlation between the amount of proteinuria and TGF-beta was not found. As has been previously demonstrated in experimental studies, TGF-beta may play a similar role in human glomerular diseases. The results obtained in this study raised the possibility that extracellular matrix might be produced by glomerular cells in vivo under the control of TGF-beta and that TGF-beta might act as a stimulator for the development of glomerulosclerosis.

Topics: Adolescent; Adult; Aged; Enzyme-Linked Immunosorbent Assay; Female; Glomerulonephritis, IGA; Glomerulonephritis, Membranous; Glomerulosclerosis, Focal Segmental; Humans; Lupus Erythematosus, Systemic; Male; Middle Aged; Proteinuria; Reference Standards; Transforming Growth Factor beta

This study was designed to investigate the mechanisms by which marine lipids rich in long chain omega-3 fatty acids inhibit autoimmune disease and prolong the survival rate in female (NZB/NZW) F1 (B/W) mice, an animal model for human SLE. Nutritionally adequate semipurified diets containing at 10% either corn oil (CO) or fish oil (FO) were fed from 1 mo of age and were monitored for proteinuria and survival. Proteinuria was detected earlier and became progressively severe in CO-fed mice. The average life span was significantly shortened by the CO diet (266.7 days +/- 12.5), whereas FO extended the survival significantly (402.1 days +/- 26.1; p < 0.001). A cross-sectional study at 6.5 mo of age revealed an increased proliferative response to T cell mitogens including bacterial superantigens and decreased serum anti-dsDNA Ab titers in the FO group compared with the CO group. Furthermore, splenocytes from the FO group when stimulated with Con A had higher IL-2 and lower IL-4 production similar to that of young (3.5 mo) mice. Flow cytometric analyses of splenocytes revealed lower Ig+, higher lymphocyte endothelial cell adhesion molecule-1, and lower Pgp-1+ cells within CD4+ and CD8+ subsets in FO-fed mice. Also, elevated IL-2 and IL-4 and significantly higher TGF-beta 1 and lower c-myc and c-ras mRNA expression and higher TGF-beta 1 and significantly lower c-Myc and c-Ha-Ras proteins were detected in spleens of FO-fed mice. Fatty acid analysis revealed significantly higher linoleic (18:2 omega-6) and arachidonic (20:4 omega-6) acid levels in splenocytes of the CO-fed group and higher eicosapentaenoic (20:5 omega-3) and docosahexanoic (22:6 omega-3) acid levels in the FO-fed group, indicating that changes in membrane fatty acid composition may contribute to the altered immune function and gene expression during the development of murine SLE.

Topics: Animals; Antibodies, Antinuclear; Autoimmune Diseases; Body Weight; Disease Models, Animal; Fatty Acids, Omega-3; Female; Gene Expression; In Vitro Techniques; Interleukin-2; Interleukin-4; Lupus Erythematosus, Systemic; Lymphocyte Activation; Lymphocyte Subsets; Mice; Mice, Inbred NZB; Oncogenes; Proteinuria; RNA, Messenger; Spleen; Transforming Growth Factor beta

The spontaneous increase in the transcription of the transforming growth factor beta (TGF beta) gene in bronchoalveolar mononuclear cells of patients with autoimmune diseases affecting the lung has been shown by northern blot assay and a nuclear run on transcription assay. Transcription of the TGF beta gene in bronchoalveolar mononuclear cells of patients with autoimmune diseases affecting the lung was increased 10 times compared with normal healthy subjects or patients with bronchial asthma as controls. This observation, confirmed by protein data, suggests that TGF beta, a potent mitogen for fibroblasts, may be produced in bronchoalveolar mononuclear cells during an active immune response in such patients and may be involved in autoimmune related changes of the pathophysiology of cytokine networks when the lung is affected, such as in lung fibrosis.

Topics: Blotting, Northern; Bronchoalveolar Lavage Fluid; Female; Fibrosis; Genetic Techniques; Humans; Lung; Lung Diseases; Lupus Erythematosus, Systemic; Male; Monocytes; RNA, Messenger; Scleroderma, Systemic; Transcription, Genetic; Transforming Growth Factor beta

The spontaneous elevation of the transcription of the transforming growth factor-beta (TGF-beta) gene in broncho-alveolar mononuclear cells (BMC) of individuals with autoimmune diseases with lung involvement, by nuclear run-on transcription assay, is shown in this study. In quantification analysis of TGF-beta gene transcription, we found more than 10 times the enhanced transcription of the TGF-beta gene in BMC of individuals with autoimmune diseases with lung involvement, in comparison to normal healthy subjects or patients with bronchial asthma used as controls. Our observation suggests that TGF-beta, a potent mitogen for fibroblasts, may be produced in BMC during an active immune response in individuals with systemic autoimmune diseases with lung involvement, and may be involved in autoimmune-related pathophysiological changes of cytokine networks in lung involvement such as lung fibrosis.

Topics: Autoimmune Diseases; Bronchoalveolar Lavage Fluid; Female; Humans; Leukocytes, Mononuclear; Lupus Erythematosus, Systemic; Male; Pulmonary Fibrosis; Scleroderma, Systemic; Transcription, Genetic; Transforming Growth Factor beta

B cells from systemic lupus erythematosus (SLE) patients have been shown to be hyperactive as measured by proliferation and immunoglobulin production. We find that B cells from 6 of 13 SLE patients, in the absence of prior activation, respond two to three times better to recombinant 12-kDa B cell growth factor (BCGF) than do normal or rheumatoid arthritis B cells (p less than 0.005). B cells from normally responsive SLE patients require an anti-mu antibody activation step to generate similar proliferative signal in response to r12-kDa-BCGF. There are no clinical or serological parameters that distinguish these hyperresponsive SLE patients from the normally responsive SLE patients. The combination of r12-kDa-BCGF and interleukin 4 (IL4) gives an enhanced response with both normal and SLE B cells. Transforming growth factor type beta (TGF-beta) suppresses the response to r12-kDa-BCGF in a dose-dependent fashion using B cells from both healthy donors and SLE patients. We conclude that peripheral blood B cells are in an activated state (as detected by response to 12-kDa-BCGF) in approximately 50% of SLE patients. These B cells respond normally to regulation by IL4 and TGF-beta. A therapeutic approach aimed at reducing the B cell hyperactivity in SLE would involve suppressing the effects of 12-kDa-BCGF and IL4 while at the same time enhancing the effects of TGF-beta.

Topics: Adolescent; Adult; Aged; B-Lymphocytes; Cells, Cultured; Female; Humans; Interleukin-2; Interleukin-4; Lupus Erythematosus, Systemic; Lymphocyte Activation; Male; Middle Aged; Recombinant Proteins; Transforming Growth Factor beta