RESUMEN
OBJECTIVE: To link changes in the B-cell transcriptome from systemic lupus erythematosus (SLE) patients with those in their macroenvironment, including cellular and fluidic components. METHODS: Analysis was performed on 363 patients and 508 controls, encompassing transcriptomics, metabolomics, and clinical data. B-cell and whole-blood transcriptomes were analysed using DESeq and GSEA. Plasma and urine metabolomics peak changes were quantified and annotated using Ceu Mass Mediator database. Common sources of variation were identified using MOFA integration analysis. RESULTS: Cellular macroenvironment was enriched in cytokines, stress responses, lipidic synthesis/mobility pathways and nucleotide degradation. B cells shared these pathways, except nucleotide degradation diverted to nucleotide salvage pathway, and distinct glycosylation, LPA receptors and Schlafen proteins. CONCLUSIONS: B cells showed metabolic changes shared with their macroenvironment and unique changes directly or indirectly induced by IFN-α signalling. This study underscores the importance of understanding the interplay between B cells and their macroenvironment in SLE pathology.
Asunto(s)
Linfocitos B , Lupus Eritematoso Sistémico , Metabolómica , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/metabolismo , Humanos , Linfocitos B/inmunología , Linfocitos B/metabolismo , Femenino , Adulto , Masculino , Transcriptoma , Persona de Mediana Edad , Perfilación de la Expresión Génica , MultiómicaRESUMEN
Abatacept mimics natural CD152 and competes with CD28 for binding to CD80/CD86 on APC, such as B cells, thereby preventing T cell activation. However, its potential impact on B cells has not been identified. The aim of this study was to assess whether abatacept can potentiate the immunoregulatory properties of B cells in vitro and in patients with rheumatoid arthritis (RA). T and B cells from healthy controls were purified. The suppressor properties of B cells in the presence of abatacept or control IgG1 were evaluated based on the ability of these cells to inhibit the polyclonal expansion (anti-CD3/CD28 stimulation) of T cells or their differentiation into Th1 or Th17 cells. Similar analyses were also performed with cells from RA patients before and 3 mo after abatacept initiation. Abatacept significantly potentiated regulatory B cell regulatory functions by enhancing their ability to produce IL-10 and TGF-ß, resulting in the increased generation of regulatory T cells and limited T cell proliferation and differentiation into Th1 and Th17 cells. Interestingly, B cells isolated from patients that received a 3-mo treatment with abatacept had an increased ability to reduce T cell functions, confirming the above observations. Abatacept binding to CD80/CD86 induces and promotes regulatory B cell functions by enhancing the ability of these cells to produce IL-10 and TGF-ß in vitro and in RA patients.
Asunto(s)
Abatacept/inmunología , Artritis Reumatoide/inmunología , Linfocitos B Reguladores/inmunología , Interleucina-10/inmunología , Células TH1/inmunología , Factor de Crecimiento Transformador beta/inmunología , Antirreumáticos/inmunología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Activación de Linfocitos/inmunologíaRESUMEN
Antibody-secreting cells (ASC) are divided into two principal subsets, including the long-lived plasma cell (PC) subset residing in the bone marrow and the short-lived subset, also called plasmablast (PB). PB are described as a proliferating subset circulating through the blood and ending its differentiation in tissues. Due to their inherent heterogeneity, the molecular signature of PB is not fully established. The purpose of this study was to decipher a specific PB signature in humans and mice through a comprehensive meta-analysis of different data sets exploring the PB differentiation in both species and across different experimental conditions. The present study used recent analyses using whole RNA sequencing in prdm1-GFP transgenic mice to define a reliable and accurate PB signature. Next, we performed similar analysis using current data sets obtained from human PB and PC. The PB-specific signature is composed of 155 and 113 genes in mouse and human being, respectively. Although only nine genes are shared between the human and mice PB signature, the loss of B-cell identity such as the down-regulation of PAX5, MS4A1, (CD20) CD22 and IL-4R is a conserved feature across species and across the different experimental conditions. Additionally, we observed that the IRF8 and IRF4 transcription factors have a specific dynamic range of expression in human PB. We thus demonstrated that IRF4/IRF8 intranuclear staining was useful to define PB in vivo and in vitro and able to discriminate between atypical PB populations and transient states.
Asunto(s)
Células Productoras de Anticuerpos/inmunología , Linfocitos B/inmunología , Células Plasmáticas/inmunología , Animales , Antígenos CD20/genética , Diferenciación Celular , Glicoproteínas/genética , Humanos , Ratones , Ratones Transgénicos/genética , Factor de Transcripción PAX5/genética , Factor 1 de Unión al Dominio 1 de Regulación Positiva/genética , Análisis de Secuencia de ARN , Transcriptoma , Secuenciación Completa del GenomaRESUMEN
The control of the activities of regulatory B (Breg) cells in immune disorders is an emerging therapeutic strategy for the recovery of immune homeostasis. Manipulating B cells using numerous drugs in vivo affect their regulatory functions, although a direct link has not yet been demonstrated. Glatiramer acetate (GA) is a synthetic polypeptide that is used in the treatment of inflammatory and autoimmune diseases. We experimented on an in vitro coculture system to determine its direct effects on the Breg cell properties of human B cells. We found that GA improves the B cell-dependent control of T cells' immune responses. When B cells are stimulated by GA, the T cell proliferation and their Th1 IFN-γ production are further inhibited, whereas the B cell production of IL-10 is further enhanced. GA binds preferentially to the memory B cells and the activation of sorted B cell subsets shows that GA-dependent increased Breg cell activities are specifically supported by the B cells' memory compartment. Moreover, we found that the defective regulations that emerge from the B cells of systemic lupus erythematosus patients can be restored by GA stimulation. Overall, these data demonstrate that GA stimulates the Breg functions mainly by shifting the memory B cells known to contribute to the T cell-dependent inflammatory response into Breg cells. Our results also indicate that GA treatment could be a useful therapy for recovering the Breg cells in autoimmune situations in which their activities are defective.
Asunto(s)
Linfocitos B Reguladores/efectos de los fármacos , Linfocitos B Reguladores/inmunología , Acetato de Glatiramer/farmacología , Inmunosupresores/farmacología , Técnicas de Cocultivo , Humanos , Lupus Eritematoso Sistémico/inmunologíaRESUMEN
Autoimmune disease development depends on multiple factors, including genetic and environmental. Abnormalities such as sialylation levels and/or quality have been recently highlighted. The adjunction of sialic acid at the terminal end of glycoproteins and glycolipids is essential for distinguishing between self and non-self-antigens and the control of pro- or anti-inflammatory immune reactions. In autoimmunity, hyposialylation is responsible for chronic inflammation, the anarchic activation of the immune system and organ lesions. A detailed characterization of this mechanism is a key element for improving the understanding of these diseases and the development of innovative therapies. This review focuses on the impact of sialylation in autoimmunity in order to determine future treatments based on the regulation of hyposialylation.
Asunto(s)
Autoanticuerpos/metabolismo , Enfermedades Autoinmunes/inmunología , Procesamiento Proteico-Postraduccional , Ácidos Siálicos/metabolismo , Animales , Autoanticuerpos/inmunología , Enfermedades Autoinmunes/terapia , Humanos , Inmunofenotipificación/métodos , Medicina de Precisión/métodos , Ácidos Siálicos/inmunologíaRESUMEN
BACKGROUND: Follicular helper T (TFH) cells support terminal B-cell differentiation. Human regulatory B (Breg) cells modulate cellular responses, but their control of TFH cell-dependent humoral immune responses is unknown. OBJECTIVE: We sought to assess the role of Breg cells on TFH cell development and function. METHODS: Human T cells were polyclonally stimulated in the presence of IL-12 and IL-21 to generate TFH cells. They were cocultured with B cells to induce their terminal differentiation. Breg cells were included in these cultures, and their effects were evaluated by using flow cytometry and ELISA. RESULTS: B-cell lymphoma 6, IL-21, inducible costimulator, CXCR5, and programmed cell death protein 1 (PD-1) expressions increased on stimulated human T cells, characterizing TFH cell maturation. In cocultures they differentiated B cells into CD138+ plasma and IgD-CD27+ memory cells and triggered immunoglobulin secretions. Breg cells obtained by Toll-like receptor 9 and CD40 activation of B cells prevented TFH cell development. Added to TFH cell and B-cell cocultures, they inhibited B-cell differentiation, impeded immunoglobulin secretions, and expanded Foxp3+CXCR5+PD-1+ follicular regulatory T cells. Breg cells modulated IL-21 receptor expressions on TFH cells and B cells, and their suppressive activities involved CD40, CD80, CD86, and intercellular adhesion molecule interactions and required production of IL-10 and TGF-ß. CONCLUSION: Human Breg cells control TFH cell maturation, expand follicular regulatory T cells, and inhibit the TFH cell-mediated antibody secretion. These novel observations demonstrate a role for the Breg cell in germinal center reactions and suggest that deficient activities might impair the TFH cell-dependent control of humoral immunity and might lead to the development of aberrant autoimmune responses.
Asunto(s)
Linfocitos B Reguladores/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos B Reguladores/fisiología , Diferenciación Celular , Células Cultivadas , Técnicas de Cocultivo , Humanos , Interleucina-12/inmunología , Interleucinas/inmunología , Linfocitos T Colaboradores-Inductores/fisiologíaRESUMEN
By their suppressive functions, regulatory B (Breg) cells are considered as key elements in the control and development of various disease states. Many signals can induce Bregs in vivo and in vitro and often from heterogeneous populations. Several specific signals delivered in a timely immunological context contribute to the establishment of Bregs. These are endogenous and physiological signals or stimuli, widely discussed in the literature participating in the establishment of an effective immune response. However, exogenous signals, much less clearly identified can also be considered as Bregs inducers. These extrinsic signals are capable of directly or indirectly influencing the suppressive capacity of Bregs, but also their expansion and functional restoration in its absence. Faced with the excitement generated by the development of processes favoring the expansion of Bregs in mice for therapeutic purposes, the challenge today is to extrapolate such approaches in humans. This perspective may already be in effect.
Asunto(s)
Antineoplásicos/farmacología , Linfocitos B Reguladores/efectos de los fármacos , Inmunosupresores/farmacología , Vitaminas/farmacología , Corticoesteroides/farmacología , Animales , Anticuerpos Monoclonales Humanizados/farmacología , Factor Activador de Células B/inmunología , Linfocitos B Reguladores/inmunología , Antígenos CD40/inmunología , Citocinas/inmunología , Humanos , Metotrexato/farmacología , Ácido Micofenólico/farmacología , Pirroles/farmacología , Quinazolinas/farmacología , Receptores de Antígenos de Linfocitos B/inmunología , Semaforinas/farmacología , Sirolimus/farmacología , Receptores Toll-Like/inmunología , Tretinoina/farmacología , Vitamina D/farmacologíaRESUMEN
In kidney transplantation, the composition of the B-cell compartment is increasingly identified as an important determinant for graft outcome. Whereas naive and transitional B cells have been associated with long-term allograft survival and operational tolerance, memory B cells have been linked to graft rejection and graft loss. Chronic antibody-mediated rejection now represents a major complication in transplantation and is a challenge in current therapeutics. Here, we show that patients with chronic antibody-mediated rejection display a unique B-cell phenotype with a reduced ratio of activated to memory B cells associated with an impaired immunosuppressive activity. The regulatory functions of the B cells depended on their maturation status. Thus, phenotypic and functional analyses of the B-cell compartment may be indicated for appropriate follow-up after transplantation and drive therapy in the establishment of transplant tolerance processes.
Asunto(s)
Linfocitos B/inmunología , Rechazo de Injerto/inmunología , Tolerancia Inmunológica , Trasplante de Riñón/efectos adversos , Adulto , Anciano , Células Cultivadas , Enfermedad Crónica , Femenino , Humanos , Memoria Inmunológica , Masculino , Persona de Mediana Edad , Linfocitos T/fisiologíaRESUMEN
Primary Sjögren's syndrome (pSS) is one of the most frequent autoimmune systemic diseases, mainly characterized by ocular and oral dryness due to the progressive destruction of lachrymal and salivary glands by an inflammatory process. A noteworthy proportion of patients also features extraglandular manifestations, sometimes severe and life-threatening. Until now, its management relies mostly on symptomatic interventions, long-term monitoring, and, in patients with severe systemic complications, immunosuppressive drugs can be provided. However, recent years have seen great progresses in the understanding of the pathological processes of the disease. The central role of regulatory lymphocytes, the implication of the type 1 interferon pathway in some patients or the importance of epigenetics have been highlighted. New classification criteria have been recently published and have shed in light an international attempt for a better recognition of the patients, probably thanks to the development of new diagnostic procedures such as salivary gland ultrasonography. To facilitate the detection of treatment efficacy in clinical trials and to help in determining which subgroups of patients would have benefits from intensive therapies, a better definition of activity scores and the availability of new prognostic markers are urgent. Thereby, the development of future therapies should be based on specific molecular signatures that will enable a personalized management of each patient. This review focuses on the most striking advances in the fields of pathophysiology, diagnosis and treatment of pSS, which generate a great hope for pSS patients.
Asunto(s)
Síndrome de Sjögren/diagnóstico , Síndrome de Sjögren/terapia , Humanos , Síndrome de Sjögren/etiologíaRESUMEN
Toll-like receptors (TLRs) are positioned at the interface between innate and adaptive immunity. Unlike others, those such as TLR9, that recognize nucleic acids, are confined to the endosomal compartment and are scarce on the cell surface. Here, we present evidence for TLR9 expression on the plasma membrane of B cells. In contrast to endosomal TLR9, cell surface TLR9 does not bind CpG-B oligodeoxynucleotides. After B cell-receptor (BCR) stimulation, TLR9 was translocated into lipid rafts with the BCR, suggesting that it could serve as a co-receptor for BCR. Nevertheless, stimulation of B cells with anti-TLR9 antibodies did not modify the BCR-induced responses despite up-regulation of tyrosine phosphorylation of proteins. However, CpG-B activation of B cells, acting synergistically with BCR signals, was inhibited by anti-TLR9 stimulation. Induction of CD25 expression and proliferation of B cells were thus down-regulated by the engagement of cell surface TLR9. Overall, our results indicate that TLR9 expressed on the plasma membrane of B cells might be a negative regulator of endosomal TLR9, and could provide a novel control by which activation of autoreactive B cells is restrained.
Asunto(s)
Linfocitos B/inmunología , Linfocitos B/metabolismo , Membrana Celular/metabolismo , Inmunomodulación , Receptor Toll-Like 9/metabolismo , Células Cultivadas , Humanos , Inmunofenotipificación , Ligandos , Activación de Linfocitos/inmunología , Oligodesoxirribonucleótidos/metabolismo , Unión ProteicaRESUMEN
Mature dendritic cells (DCs) are stimulators of T-cell immune response, whereas immature DCs support T-cell tolerance. Murine B cells can inhibit the production of IL-12 by DCs and thereby hinder the inflammatory response. Notwithstanding the importance of this modulation, only a few studies are available in humans. Here, we have developed an in vitro model of cocultures to assess its significance. We establish that human activated B cells restrained the development of monocytes into immature DCs and their differentiation into mature DCs. In addition, they decreased the density of HLA-DR from mature DCs, the expression of CD80 and CD86 coactivation molecules, the production of IL-12p70 required for antigen presentation and Th1 differentiation, and inhibited the DC-induced T-cell proliferation. These modulations were mediated by CD19(+)IgD(low)CD38(+)CD24(low)CD27(-) B cells and needed direct cell-to-cell contacts that involved CD62L for the control of CD80 and CD86 expression and a soluble factor for the control of IL-12 production. Moreover, mature DCs from patients with systemic lupus erythematosus displayed insensitivity to the regulation of IL-12. Overall, it appears that human B cells can regulate DC maturation and function and that inefficient B-cell regulation may influence an improper balance between an effector inflammatory response and tolerance induction.
Asunto(s)
Presentación de Antígeno/inmunología , Artritis Reumatoide/inmunología , Linfocitos B/inmunología , Células Dendríticas/inmunología , Lupus Eritematoso Sistémico/inmunología , Monocitos/inmunología , Linfocitos T/inmunología , Animales , Artritis Reumatoide/metabolismo , Linfocitos B/metabolismo , Estudios de Casos y Controles , Diferenciación Celular , Células Cultivadas , Técnicas de Cocultivo , Células Dendríticas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Antígenos HLA-DR/inmunología , Humanos , Tolerancia Inmunológica , Interleucina-12/metabolismo , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Lupus Eritematoso Sistémico/metabolismo , Activación de Linfocitos , Ratones , Tonsila Palatina , Linfocitos T Reguladores/inmunologíaRESUMEN
The recent emergence of imaging mass cytometry technology has led to the generation of an increasing amount of high-dimensional data and, with it, the need for suitable performant bioinformatics tools dedicated to specific multiparametric studies. The first and most important step in treating the acquired images is the ability to perform highly efficient cell segmentation for subsequent analyses. In this context, we developed YOUPI (Your Powerful and Intelligent tool) software. It combines advanced segmentation techniques based on deep learning algorithms with a friendly graphical user interface for non-bioinformatics users. In this article, we present the segmentation algorithm developed for YOUPI. We have set a benchmark with mathematics-based segmentation approaches to estimate its robustness in segmenting different tissue biopsies.
Asunto(s)
Algoritmos , Programas Informáticos , Citometría de ImagenRESUMEN
Maturation of B cells depends on environmental stimuli. Peripheral immature B cells develop into follicular pathway when antigenic stimulation is combined with T cell signals. Here, we wished to identify stimuli contributing to the development into marginal zone B cells known to be involved in autoimmune response. We found that TLR9 stimulation of transitional B cells induces proliferation and specific maturation into CD24(-) CD38(+) CD21(high) CD23(low) IgM(high) IgD(low) and Notch2(high) B cells characteristics of marginal zone B cells. Terminal differentiation into antibody-secreting cell associated with isotype switch commitment is also triggered which leads to a striking production of autoantibodies. Interestingly, mature B cells do not differentiate into marginal zone pathway following TLR9 stimulation, nor do transitional B cells under antigenic and T cell combined signals. These results suggest that transitional B cells are specifically sensitive to TLR9 stimulation to produce autoreactive marginal zone B cells.
Asunto(s)
Células Productoras de Anticuerpos/inmunología , Autoanticuerpos/biosíntesis , Autoinmunidad , Células Precursoras de Linfocitos B/inmunología , Receptor Toll-Like 9/metabolismo , Adyuvantes Inmunológicos/farmacología , Células Productoras de Anticuerpos/citología , Células Productoras de Anticuerpos/efectos de los fármacos , Antígenos CD/inmunología , Autoanticuerpos/inmunología , Diferenciación Celular , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Femenino , Sangre Fetal/citología , Sangre Fetal/efectos de los fármacos , Sangre Fetal/inmunología , Citometría de Flujo , Humanos , Activación de Linfocitos/efectos de los fármacos , Oligodesoxirribonucleótidos/farmacología , Tonsila Palatina/citología , Tonsila Palatina/efectos de los fármacos , Tonsila Palatina/inmunología , Células Precursoras de Linfocitos B/citología , Células Precursoras de Linfocitos B/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Transducción de Señal/inmunología , Linfocitos T/citología , Linfocitos T/inmunología , Receptor Toll-Like 9/inmunologíaRESUMEN
Be they follicular cells within the germinal centers (GCs) or marginal zone (MZ), all naïve mature B lymphocytes need tonic signaling to stay alive. We reasoned that the same holds true for those B lymphocytes that proliferate in the salivary glands (SGs) of patients with primary Sjögren's syndrome. Based on B cell infiltration, 11 SGs and three tonsil samples were selected for further examination. Tissue sections were stained using CD20 combined with CD10, CD21, CD27, CD38 or IgD. They were also laser-microdissected for quantitative RT-PCR of transcription factors, GC-specific activation-induced cytidine deaminase (AID) and TLR9. Some B cell aggregates proved to be real GCs according to their membrane markers, whereas others were clusters of transitional type II B cells. These contained mRNAs for Notch-2 and Blimp-1, but not for Pax-5, Bcl-6 and AID. Unanticipated was the finding of mRNAs for TLR9 in these clusters of MZ B-cells, but not in the real GCs. Not only do TLR9 deliver sufficiency of tonic signaling to keep B cells alive, but they also confer autoreactive B cells with an MZ-like phenotype. Thus, TLRs might be targets for forthcoming biotherapies.
Asunto(s)
Linfocitos B/inmunología , Glándulas Salivales Menores/inmunología , Síndrome de Sjögren/inmunología , Receptor Toll-Like 9/metabolismo , ADP-Ribosil Ciclasa 1/análisis , Adolescente , Adulto , Anciano , Antígenos CD20/análisis , Subgrupos de Linfocitos B/citología , Subgrupos de Linfocitos B/inmunología , Niño , Preescolar , Citidina Desaminasa/biosíntesis , Femenino , Centro Germinal/metabolismo , Humanos , Inmunoglobulina D/análisis , Persona de Mediana Edad , Tonsila Palatina , Factor 1 de Unión al Dominio 1 de Regulación Positiva , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Complemento 3d/análisis , Receptores de IgG/análisis , Proteínas Represoras/biosíntesis , Glándulas Salivales Menores/metabolismo , Síndrome de Sjögren/metabolismo , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/análisisRESUMEN
The potential predictive value of tumor bulk, genetic, and immunological variants in patients with low-grade non-Hodgkin's lymphoma to respond to treatment with rituximab (RTX) monotherapy was evaluated. Thus, the value of assessing the effect of 18-fluoro-desoxy-D-glucose (FDG) uptake on PET scan, polymorphisms in Fc gamma receptor (FcγR) IIIa-158, FcγRIIa-131, and C1qA-276 genes in predicting the response to treatment were evaluated in 50 low-grade non-Hodgkin's lymphoma patients. The influence of RTX pharmacokinetics, plasma levels of the B cell-activating factor (BAFF), and human antichimeric antibodies was also investigated. The therapeutic response was evaluated 10 weeks after treatment using revised Cheson's criteria. Lower maximal standardized uptake values (SUV(max)) at baseline were predictive of complete response. FcγRIIIa-158 polymorphism was also associated with complete response to RTX confirming previous findings, whereas polymorphisms in the FcγRIIa-131 and C1qA-276 genes were not. Lower blood levels of RTX were observed in males, but the effectiveness of RTX in males and females was the same. BAFF was not detectable in plasma before or after treatment, and no patients developed human antichimeric antibodies. Low-grade non-Hodgkin's lymphoma patients with a low SUV(max) at baseline and an FcγRIIIa-158 V/V genotype generally had a complete response to RTX.
Asunto(s)
Anticuerpos Monoclonales de Origen Murino/uso terapéutico , Antineoplásicos/uso terapéutico , Linfoma de Células B/diagnóstico , Linfoma de Células B/tratamiento farmacológico , Adulto , Anciano , Anticuerpos Monoclonales de Origen Murino/farmacocinética , Antineoplásicos/farmacocinética , Factor Activador de Células B/sangre , Complemento C1q/genética , Femenino , Humanos , Masculino , Persona de Mediana Edad , Imagen Multimodal , Clasificación del Tumor , Norleucina/análogos & derivados , Norleucina/sangre , Polimorfismo Genético , Tomografía de Emisión de Positrones , Pronóstico , Receptores de IgG/genética , Rituximab , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Carga TumoralRESUMEN
During the antiphospholipid syndrome, beta2-gpI interacts with phospholipids on endothelial cell (EC) surface to allow the binding of autoantibodies. However, induced-pathogenic intracellular signals suggest that beta2-gpI associates also with a receptor that is still not clearly identified. TLR2 and TLR4 have long been suspected, yet interactions between TLRs and beta2-gpI have never been unequivocally proven. The aim of the study was to identify the TLR directly involved in the binding of beta2-gpI on EC surface. beta2-gpI was not synthesized and secreted by ECs in vitro, but rather taken up from FCS. This uptake occurred through association with TLR2 and TLR4 which partitioned together in the lipid rafts of ECs. After coimmunoprecipitation, mass-spectrometry identification of peptides demonstrated that TLR2, but not TLR4, was implicated in the beta2-gpI retention. These results were further confirmed by plasmon resonance-based studies. Finally, siRNA were used to obtain TLR2-deficient ECs that lost their ability to bind biotinylated beta2-gpI and to trigger downstream phosphorylation of kinases and activation of NFkappaB. TLR4 may upregulate TLR2 expression, thereby contributing to beta2-gpI uptake. However, our data demonstrate that direct binding of beta2-gpI on EC surface occurs through direct interaction with TLR2. Furthermore, signaling for anti-beta2-gpI may be envisioned as a multiprotein complex concentrated in lipid rafts on the EC membrane.
Asunto(s)
Endotelio Vascular/metabolismo , Receptor Toll-Like 2/metabolismo , beta 2 Glicoproteína I/metabolismo , Secuencia de Aminoácidos , Animales , Bovinos , Línea Celular , Células Hep G2 , Humanos , Inmunoprecipitación , Microdominios de Membrana/inmunología , Microdominios de Membrana/metabolismo , Datos de Secuencia Molecular , Péptidos/aislamiento & purificación , Péptidos/metabolismo , Unión Proteica/inmunología , Receptor Toll-Like 2/biosíntesis , Receptor Toll-Like 2/aislamiento & purificación , Receptor Toll-Like 4/biosíntesis , Receptor Toll-Like 4/aislamiento & purificación , Receptor Toll-Like 4/metabolismo , Células U937 , beta 2 Glicoproteína I/biosíntesisRESUMEN
OBJECTIVE: Anti-Ro autoantibodies are among the most frequently detected extractable nuclear antigen autoantibodies, mainly associated with primary Sjögren's syndrome (SS), systemic lupus erythematosus (SLE), and undifferentiated connective tissue disease (UCTD). This study was undertaken to determine if there is a common signature for all patients expressing anti-Ro 60 autoantibodies regardless of their disease phenotype. METHODS: Using high-throughput multiomics data collected from the cross-sectional cohort in the PRECISE Systemic Autoimmune Diseases (PRECISESADS) study Innovative Medicines Initiative (IMI) project (genetic, epigenomic, and transcriptomic data, combined with flow cytometry data, multiplexed cytokines, classic serology, and clinical data), we used machine learning to assess the integrated molecular profiling of 520 anti-Ro 60+ patients compared to 511 anti-Ro 60- patients with primary SS, patients with SLE, and patients with UCTD, and 279 healthy controls. RESULTS: The selected clinical features for RNA-Seq, DNA methylation, and genome-wide association study data allowed for a clear distinction between anti-Ro 60+ and anti-Ro 60- patients. The different features selected using machine learning from the anti-Ro 60+ patients constituted specific signatures when compared to anti-Ro 60- patients and healthy controls. Remarkably, the transcript Z score of 3 genes (ATP10A, MX1, and PARP14), presenting with overexpression associated with hypomethylation and genetic variation and independently identified using the Boruta algorithm, was clearly higher in anti-Ro 60+ patients compared to anti-Ro 60- patients regardless of disease type. Our findings demonstrated that these signatures, enriched in interferon-stimulated genes, were also found in anti-Ro 60+ patients with rheumatoid arthritis and those with systemic sclerosis and remained stable over time and were not affected by treatment. CONCLUSION: Anti-Ro 60+ patients present with a specific inflammatory signature regardless of their disease type, suggesting that a dual therapeutic approach targeting both Ro-associated RNAs and anti-Ro 60 autoantibodies should be considered.
Asunto(s)
Enfermedades Autoinmunes , Lupus Eritematoso Sistémico , Síndrome de Sjögren , Enfermedades Indiferenciadas del Tejido Conectivo , Anticuerpos Antinucleares , Antígenos Nucleares , Autoanticuerpos , Autoantígenos , Enfermedades Autoinmunes/genética , Estudios Transversales , Citocinas , Estudio de Asociación del Genoma Completo , Humanos , Interferones , Lupus Eritematoso Sistémico/genética , Aprendizaje Automático , Ribonucleoproteínas/genética , Síndrome de Sjögren/genéticaRESUMEN
Systemic lupus erythematosus (SLE) and primary Sjögren's syndrome (pSS) are two autoimmune diseases characterised by the production of pathogenic autoreactive antibodies. Their aetiology is poorly understood. Nevertheless, they have been shown to involve several factors, such as infections and epigenetic mechanisms. They also likely involve a physiological process known as glycosylation. Both SLE T cell markers and pSS-associated autoantibodies exhibit abnormal glycosylation. Such dysregulation suggests that defective glycosylation may also occur in B cells, thereby modifying their behaviour and reactivity. This study aimed to investigate B cell subset glycosylation in SLE, pSS and healthy donors and to extend the glycan profile to serum proteins and immunoglobulins. We used optimised lectin-based tests to demonstrate specific glycosylation profiles on B cell subsets that were specifically altered in both diseases. Compared to the healthy donor B cells, the SLE B cells exhibited hypofucosylation, whereas only the pSS B cells exhibited hyposialylation. Additionally, the SLE B lymphocytes had more galactose linked to N-acetylglucosamine or N-acetylgalactosamine (Gal-GlcNAc/Gal-GalNAc) residues on their cell surface markers. Interestingly, some similar alterations were observed in serum proteins, including immunoglobulins. These findings indicate that any perturbation of the natural glycosylation process in B cells could result in the development of pathogenic autoantibodies. The B cell glycoprofile can be established as a preferred biomarker for characterising pathologies and adapted therapeutics can be used for patients if there is a correlation between the extent of these alterations and the severity of the autoimmune diseases.
Asunto(s)
Enfermedades Autoinmunes , Lupus Eritematoso Sistémico , Autoanticuerpos , Autoinmunidad , Linfocitos B/metabolismo , Glicosilación , HumanosRESUMEN
Regulatory functions for B lymphocytes have been reported in murine models of autoimmune diseases in which B-cell deficient mice were shown to exhibit exacerbated disease. The B cells responsible for the immune regulations were identified as a subpopulation of interleukin 10-secreting cells. However, the mechanism of induction and the characteristics of regulatory B cells in humans have been hardly studied. This study reports that regulation of T cell responses can be induced by B cells following CD40-dependent cognate interaction. T cell proliferation and cytokine production were differentially regulated. Thus, CD40-induced regulatory B cells partially inhibited T cell proliferation following CD40 interaction without requirement of soluble factor. In contrast, modulation of Th1 differentiation resulted from CD80- and CD86-dependent interactions and from IL-10 production. The suppressive effects were mediated by CD19(high)IgD+CD38(high)CD24(high)CD5(high) B cells and appeared to be indirect, through the induction of regulatory T cells as indicated by the appearance of Foxp3+CD4+CD25+T cells. These data suggest that activation signals from T cells initiate regulatory properties in B cells that modulate T cell responses involving regulatory T cells. Finally, studies in autoimmune patients revealed that regulation of T cell proliferation was defective in systemic lupus erythematosus but efficient in other diseases. Restoration of efficient B-cell regulatory activity could provide innovative B-cell based treatment of autoimmune diseases.
Asunto(s)
Linfocitos B/inmunología , Activación de Linfocitos , Linfocitos T/fisiología , Antígenos CD/fisiología , Antígeno B7-1/fisiología , Antígenos CD40/fisiología , Ligando de CD40/fisiología , Células Cultivadas , Humanos , Interleucina-10/biosíntesis , Lupus Eritematoso Sistémico/inmunología , Linfocitos T Reguladores/fisiología , Tetraspanina 28RESUMEN
One way for intravenous Ig (IVIg) to affect responses of the B cells might be to operate through their TLR7 and TLR9. We confirm the ability of TLR agonists to induce CD25 expression in B cells. For this to occur, sialylated Fc-gamma of IgG included in the IVIg preparation are required. As a result, IVIg suppresses TLR-induced production of the proinflammatory IL-6, but not that of the anti-inflammatory IL-10. That is, IVIg mimics the effects of the MyD88 inhibitor. Finally, as we previously showed that IVIg induces CD22 to recruit the inhibitory SHP-1, we established that this enzyme was also involved in IVIg-induced inhibition of TLR9 signaling. This is the first report to demonstrate such a mechanism underlying the negative impact of IVIg on B lymphocytes.