Autophagy and rheumatoid arthritis: Current knowledges and future perspectives

Autophagy is a degradation mechanism by which cells recycle cytoplasmic components to generate energy. By influencing lymphocyte development, survival, and proliferation, autophagy regulates the immune responses against self and non-self antigens. Deregulation of autophagic pathway has recently been implicated in the pathogenesis of several autoimmune diseases, including rheumatoid arthritis (RA). Indeed, autophagy seems to be involved in the generation of citrullinated peptides, and also in apoptosis resistance in RA. In this review, we summarize the current knowledge on the role of autophagy in RA and discuss the possibility of a clinical application of autophagy modulation in this disease

Low expression of estrogen receptor β in T lymphocytes and high serum levels of anti-estrogen receptor α antibodies impact disease activity in female patients with systemic lupus erythematosus

BACKGROUND: Current evidence indicates that estrogens, in particular 17β-estradiol (E2), play a crucial role in the gender bias of autoimmune diseases although the underlying molecular mechanisms have not yet been fully elucidated. Immune cells have estrogen receptors (ERs), i.e., ERα and ERβ, that play pro- and anti-inflammatory functions, respectively, and the presence of one estrogen receptor (ER) subtype over the other might change estrogen effects, promoting or dampening inflammation. In this study, we contributed to define the influences of E2 on T cells from female patients with systemic lupus erythematosus (SLE), a representative autoimmune disease characterized by a higher prevalence in women than in men (female/male ratio 9:1). Particularly, our aim was to evaluate whether alterations of ERα and ERβ expression in T cells from female SLE patients may impact lymphocyte sensitivity to E2 and anti-ERα antibody (anti-ERα Ab) stimulation interfering with cell signaling and display a direct clinical effect. METHODS: Sixty-one premenopausal female patients with SLE and 40 age-matched healthy donors were recruited. Patients were divided into two groups based on the SLE Disease Activity Index 2000 (SLEDAI-2K) (i.e., <6 and ≥6). ER expression was evaluated in T lymphocytes by flow cytometry, immunofluorescence, and Western blot analyses. Serum anti-ERα Ab levels were analyzed by enzyme-linked immunosorbent assay (ELISA). ER-dependent signaling pathways were measured by a phosphoprotein detection kit. RESULTS: Intracellular ERβ expression was significantly lower in T cells from patients with SLEDAI-2K ≥6 as compared with healthy donors and patients with SLEDAI-2K <6 and negatively correlated with disease activity. The expression of intracellular and membrane-associated-ERα was similar in SLE and control T cells. ER-dependent signaling pathways were activated in T cells from SLE patients with SLEDAI-2K ≥6, but not with SLEDAI-2K <6, when both membrane and intracellular ERs were stimulated by co-treatment with E2 and anti-ERα Abs. CONCLUSIONS: Our results demonstrate an altered ER profile in SLE patients, possibly contributing to SLE pathogenesis and interfering with clinical activity, and highlight the potential exploitation of T cell-associated ERβ as a biomarker of disease activity

The role of dietary sodium intake on the modulation of T helper 17 cells and regulatory T cells in patients with rheumatoid arthritis and systemic lupus erythematosus

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

TNFα expressed on the surface of microparticles modulates endothelial cell fate in rheumatoid arthritis

Background: Rheumatoid arthritis (RA) is associated with a high prevalence of atherosclerosis. Recently increased levels of microparticles (MPs) have been reported in patients with RA. MPs could represent a link between autoimmunity and endothelial dysfunction by expressing tumor necrosis factor alpha (TNFα), a key cytokine involved in the pathogenesis of RA, altering endothelial apoptosis and autophagy. The aim of this study was to investigate TNFα expression on MPs and its relationship with endothelial cell fate. Methods: MPs were purified from peripheral blood from 20 healthy controls (HC) and from 20 patients with RA, before (time (T)0) and after (T4) 4-month treatment with etanercept (ETA). Surface expression of TNFα was performed by flow cytometry analysis. EA.hy926 cells, an immortalized endothelial cell line, were treated with RA-MPs purified at T0 and at T4 and also, with RA-MPs in vitro treated with ETA. Apoptosis and autophagy were then evaluated. Results: RA-MPs purified at T0 expressed TNFα on their surface and this expression significantly decreased at T4. Moreover, at T0 RA-MPs, significantly increased both apoptosis and autophagy levels on endothelial cells, in a dose-dependent manner. RA-MPs did not significantly change these parameters after 4 months of in vivo treatment with ETA. Conclusions: Our data demonstrate that MPs isolated from patients with RA exert a pathological effect on endothelial cells by TNFα expressed on their surface. In vivo and in vitro treatment with ETA modulates this effect, suggesting anti-TNF therapy protects against endothelial damage in patients with RA

Prevalence, sensitivity and specificity of antibodies against carbamylated proteins in a monocentric cohort of patients with rheumatoid arthritis and other autoimmune rheumatic diseases

Antibodies against carbamylated proteins (anti-CarP) have been recently identified in the sera of patients with rheumatoid arthritis (RA). The objective of the study was to evaluate the prevalence, sensitivity and specificity of anti-CarP compared to anti-citrullinated peptide antibodies (ACPA) and rheumatoid factor (RF), replicating the existing data in a large cohort of Italian patients with RA and extending the evaluation to other autoimmune rheumatic diseases (AIRDs)

Identification of subclinical lung involvement in ACPA-positive subjects through functional assessment and serum biomarkers

Lung involvement is related to the natural history of anti-citrullinated proteins antibodies (ACPA)-positive rheumatoid arthritis (RA), both during the pathogenesis of the disease and as a site of disease-related injury. Increasing evidence suggests that there is a subclinical, early lung involvement during the course of the disease, even before the onset of articular manifestations, which can potentially progress to a symptomatic interstitial lung disease. To date, reliable, noninvasive markers of subclinical lung involvement are still lacking in clinical practice. The aim of this study is to evaluate the diagnostic potential of functional assessment and serum biomarkers in the identification of subclinical lung involvement in ACPA-positive subjects. Fifty ACPA-positive subjects with or without confirmed diagnosis of RA (2010 ARC-EULAR criteria) were consecutively enrolled. Each subject underwent clinical evaluation, pulmonary function testing (PFT) with assessment of diffusion lung capacity for carbon monoxide (DLCO), cardiopulmonary exercise testing (CPET), surfactant protein D (SPD) serum levels dosage and high-resolution computed tomography (HRCT) of the chest. The cohort was composed of 21 ACPA-positive subjects without arthritis (ND), 10 early (disease duration &lt; 6 months, treatment-naïve) RA (ERA) and 17 longstanding (disease duration &lt; 36 months, on treatment) RA (LSRA). LSRA patients had a significantly higher frequency of overall HRCT abnormalities compared to the other groups (p = 0.001). SPD serum levels were significantly higher in ACPA-positive subjects compared with healthy controls (158.5 ± 132.3 ng/mL vs 61.27 ± 34.11 ng/mL; p &lt; 0.0001) and showed an increasing trend from ND subjects to LSRD patients (p = 0.004). Patients with HRCT abnormalities showed significantly lower values of DLCO (74.19 ± 13.2% pred. vs 131.7 ± 93% pred.; p = 0.009), evidence of ventilatory inefficiency at CPET and significantly higher SPD serum levels compared with subjects with no HRCT abnormalities (213.5 ± 157.2 ng/mL vs 117.7 ± 157.3 ng/mL; p = 0.018). Abnormal CPET responses and higher SPD levels were also associated with specific radiological findings. Impaired DLCO and increased SPD serum levels were independently associated with the presence of HRCT abnormalities. Subclinical lung abnormalities occur early in RA-associated autoimmunity. The presence of subclinical HRCT abnormalities is associated with several functional abnormalities and increased SPD serum levels of SPD. Functional evaluation through PFT and CPET, together with SPD assessment, may have a diagnostic potential in ACPA-positive subjects, contributing to the dentification of those patients to be referred to HRCT scan

Autophagy hijacking in PBMC From COVID-19 patients results in lymphopenia

Autophagy is a homeostatic process responsible for the self-digestion of intracellular components and antimicrobial defense by inducing the degradation of pathogens into autophagolysosomes. Recent findings suggest an involvement of this process in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. However, the role of autophagy in the immunological mechanisms of coronavirus disease 2019 (COVID-19) pathogenesis remains largely unexplored. This study reveals the presence of autophagy defects in peripheral immune cells from COVID-19 patients. The impairment of the autophagy process resulted in a higher percentage of lymphocytes undergoing apoptosis in COVID-19 patients. Moreover, the inverse correlation between autophagy markers levels and peripheral lymphocyte counts in COVID-19 patients confirms how a defect in autophagy might contribute to lymphopenia, causing a reduction in the activation of viral defense. These results provided intriguing data that could help in understanding the cellular underlying mechanisms in COVID-19 infection, especially in severe forms

CD4 T lymphocyte autophagy is upregulated in the salivary glands of primary Sjögren’s syndrome patients and correlates with focus score and disease activity

Background: Primary Sjögren’s syndrome (pSS) is a common chronic autoimmune disease characterized by lymphocytic infiltration of exocrine glands and peripheral lymphocyte perturbation. In the current study, we aimed to investigate the possible pathogenic implication of autophagy in T lymphocytes in patients with pSS. Methods: Thirty consecutive pSS patients were recruited together with 20 patients affected by sicca syndrome a nd/or chronic sialoadenitis and 30 healthy controls. Disease activity and damage were evaluated according to SS disease activity index, EULAR SS disease activity index, and SS disease damage index. T lymphocytes were analyzed for the expression of autophagy-specific markers by biochemical, molecular, and histological assays in peripheral blood and labial gland biopsies. Serum interleukin (IL)-23 and IL-21 levels were quantified by enzyme-linked immunosorbent assay. Results: Our study provides evidence for the first time that autophagy is upregulated in CD4+ T lymphocyte salivary glands from pSS patients. Furthermore, a statistically significant correlation was detected between lymphocyte autophagy levels, disease activity, and damage indexes. We also found a positive correlation between autophagy enhancement and the increased salivary gland expression of IL-21 and IL-23, providing a further link between innate and adaptive immune responses in pSS. Conclusions: These findings suggest that CD4+ T lymphocyte autophagy could play a key role in pSS pathogenesis. Additionally, our data highlight the potential exploitation of T cell autophagy as a biomarker of disease activity and provide new ground to verify the therapeutic implications of autophagy as an innovative drug target in pSS

AULA EN LÍNEA REDISEÑO DE PRÁCTICAS EDUCATIVAS PARA ENTORNOS VIRTUALES

En este libro digital se presentan 19 propuestas surgidas de la actividad transversal “Territorio de encuentros” del curso “Aula en línea. Prácticas educativas en entornos virtuales de aprendizaje. Postítulo de perfeccionamiento docente”, dirigido a docentes de formación docente en modalidad semipresencial, en su primera edición (2017). Se trata de la justificación documentada de conceptos y prácticas que articulan la enseñanza y el aprendizaje en ambientes educativos virtuales. El curso “Aula en línea” tiene por objetivo fortalecer la formación docente para potenciar las capacidades de captación, formación y seguimiento de estudiantes de formación docente que optan por la modalidad semipresencial. Es producto de una acción conjunta entre el Consejo de Formación en Educación (CFE), FLACSO Uruguay, El Abrojo y Fundación Telefónica-Movistar. Un gran desafío de la educación en entornos virtuales es la calidad de las propuestas. Cuando se integran tecnologías, varios son los riesgos que se corren. Como afirma Onrubia (2005:9)1 algunos de los modelos de e-learning se centran fundamentalmente en la provisión y distribución de contenidos, presuponiendo una correspondencia lineal entre lo que se enseña y lo que se aprende, ignorando tanto el rol de la actividad del sujeto que aprende, como aquella producto de la interacción y, más aún, no considerando la importancia de la ayuda pedagógica de los sujetos en esa actividad conjunta de construcción del conocimiento. Por último, no reparan en la diferencia entre diseño y uso, es decir, la diferencia entre lo que se planifica y lo que los participantes realmente acaban haciendo en el proceso de desarrollo de ese diseño. Ya en el informe Horizon 2010 del Proyecto Horizon del New Media Consortium (NMC) se señalaba que la abundancia de recursos y su facilidad para propiciar el acceso a la información, debía llevar al replanteo de las funciones docentes y su rol en las instituciones educativas insertas en la cultura digital. La función docente trasciende cada vez más la mera transmisión de información, ya que ésta se encuentra accesible en variedad de espacios digitales y a través de distintos espacios de comunicación. Esto nos sitúa en un nuevo contexto que implica el desarrollo de nuevas competencias: buscar, seleccionar y evaluar la credibilidad de la información, así como considerar los procesos vinculados a la gestión de aula. Surge la necesidad de un cambio de paradigma que se profundiza con el uso de las tecnologías para enriquecer y hacer eficiente el proceso de aprendizaje. El rediseño de las prácticas se enfoca en el aprendizaje del estudiantado y la capacitación de los docentes tutores, se adecua a la enseñanza y el aprendizaje en entornos virtuales, en especial de quienes recién comienzan. El curso pone en situación de aprendizaje a los participantes en cuanto al diseño, desarrollo y valoración de prácticas educativas, pero, en especial, promueve la reflexión sobre su propia práctica docente ya que se trata de profesionales en servicio. Favorece el desarrollo de nuevas competencias y nuevos modelos de trabajo para hacer frente a la nueva realidad tecnológica y pedagógica de la era digital. Estas competencias se adquieren a través de la resolución de tareas, dado que la resolución de la tarea es lo que hace que una persona utilice adecuadamente todos los recursos conceptuales y prácticos de los que dispone. Y las competencias se evalúan, a través de esas tareas realizadas, mediante la valoración de las evidencias generadas. Al proponer la actividad durante el curso, partimos de la premisa de que el análisis de ejemplos de buenas prácticas reforzaría el diseño de las actividades de aprendizaje. Y en especial, que la mirada sobre los 2 https://www.nmc.org/nmc-horizon/ AULA EN LÍNEA: REDISEÑO DE PRÁCTICAS EDUCATIVAS PARA ENTORNOS VIRTUALES | 11 aspectos clave de los aprendizajes, facilitaría la reflexión y la creación de los lazos necesarios entre la teoría y la práctica. Por ser una actividad que tuvo algunos pasos individuales y otros grupales, se optimizaron las interacciones a partir de la identificación, visibilidad y apropiación de aquellas acciones que pudieran ser consideradas buenas prácticas. Muchos son los investigadores que han tratado los modelos de buenas prácticas y afirman que su conocimiento y divulgación, constituye una de las opciones que permite apoyar la integración real de las tecnologías en los procesos didácticos y cognitivos a través de la educación virtual (De Pablos y Jiménez, 2007). Las buenas prácticas docentes en la educación virtual, que buscan mejorar el desempeño de un proceso, fueron propuestas en el año 1998 por la University for Industry en el Reino Unido como un modelo original e innovador (Stephenson, 2005). Sabemos que las buenas prácticas no pueden extrapolarse de forma masiva y automática. Son necesarios procesos de rediseño: adaptación, readecuación y apropiación por parte de las comunidades educativas. El proceso de interacción entre pares, que incluye la observación, el análisis, la valoración y la replicación, es un modo de aprendizaje válido. La posibilidad de contemplar su complejidad, interrelacionar la práctica con la teoría, permite repensarlas y avanzar en un nuevo diseño. De allí el motivo por el cual la difusión de las experiencias también adquiere centralidad. La actividad se planteó en pasos que contemplan actividades individuales y grupales de selección, análisis y puesta en común de siete patrones seleccionados como buenas prácticas para entornos virtuales de aprendizaje, con el propósito de reescribir las actividades de aprendizaje a partir de la adopción de estas buenas prácticas. Esto permitió a los docentes deconstruir, construir y/o reconstruir sus propias propuestas. Esperamos sean cambios genuinos y duraderos en sus “patrones conductuales” (Durán Rodríguez y Estay-Niculcar, 20165), cambios en su pensamiento y conocimiento pedagógico en que se sustentan, cambios al fin, en la enseñanza en un escenario educativo virtual, que debería ser modélico ya que es la práctica el verdadero punto de referencia en la formación del estudiante. En este trabajo se difunde una selección realizada por un Comité de Evaluación interinstitucional, con 19 propuestas que fueron producto de este proceso y se propone, a la vez, como aporte a la comunidad docente, especialmente a aquella vinculada a la formación docente semipresencial y virtual.Consejo de Formación en Educación (CFE), Fundación Telefónica Uruguay, Movistar, El Abroj