Pathogenesis of Systemic Sclerosis: pro-inflammatory role of ET-1 receptors

L'endotelina-1 (ET-1) gioca un ruolo fondamentale nella vasocostrizione, nella fibrosi e nell\u2019infiammazione, tre aspetti fondamentali nella patogenesi della sclerosi sistemica (SSc) . I recettori dell\u2019ET-1 sono ETA ed ETB. Essi sono espressi sulla maggioranza delle cellule coinvolte nella patogenesi della SSc, come le cellule endoteliali, le cellule muscolari lisce e i fibroblasti. Poco si conosce riguardo l'espressione dei recettori dell\u2019ET-1 sui leucociti. A eccezione dei macrofagi e dei monociti, non ci sono informazioni sull'espressione di ETA e ETB su linfociti, neutrofili e le altre cellule coinvolte nella risposta immunitaria innata e acquisita. Gli antagonisti dei recettori dell\u2019ET-1 sono utilizzati nel trattamento di pazienti con SSc e ulcere digitali e/o con ipertensione arteriosa polmonare. Essi hanno effetti benefici sulla vasocostrizione e la fibrosi, ma poco noto \ue8 il loro ruolo nell\u2019infiammazione. Abbiamo pertanto deciso di studiare il ruolo dell\u2019ET-1 nell\u2019infiammazione in pazienti affetti da SSc. Poich\ue9 le cellule T e B, i monociti e i neutrofili sono tra le cellule pi\uf9 importanti nelle risposta infiammatoria che si osserva nella SSc , abbiamo studiato l\u2019espressione di ETA ed ETB su queste cellule con citometria a flusso, valutando inoltre la presenza dell\u2019mRNA codificante per ETA ed ETB sulle cellule T CD4+ e sui neutrofili mediante RT-PCR . Abbiamo studiato la diversa espressione dei recettori dell\u2019ET-1 sui linfociti T e B e sui monociti di pazienti e controlli, nonch\ue9 la correlazione tra la loro espressione e alcune caratteristiche della malattia. Abbiamo inoltre valutato la modulazione dei recettori dell\u2019ET-1 sulle cellule T attivate. Al fine di valutare gli effetti pro-infiammatori dell\u2019ET-1 e il ruolo antinfiammatorio del Bosentan, antagonista dei recettori dell\u2019ET-1, abbiamo osservato come ET-1 influenzi la produzione di IFN-\u3b3 e IL-4 nelle cellule CD4+ T, in presenza o assenza del blocco recettoriale. Abbiamo studiato anche l'espressione degli mRNA codificanti per IFN-\u3b3, IL-4, IL-6, IL-10 e IL-17 sulle cellule CD4+ T mediante Real Time PCR in tempi diversi per meglio valutare in queste cellule il timing di risposta allo stimolo. Abbiamo osservato che le cellule T e i monociti esprimono sia ETA sia ETB. Abbiamo convalidato nostri dati su una coorte di pazienti e controlli. Abbiamo confermato che non solo le cellule T e i monociti, ma anche i linfociti B e i neutrofili esprimono ETA ed ETB sulla loro superficie. Abbiamo visto che l'espressione di ETA \ue8 maggiore di quella di ETB sia per quanto riguarda i linfociti T che i monociti di pazienti e controlli, mentre per le cellule B non c'\ue8 alcuna differenza di espressione. Anche i neutrofili esprimono sia ETA sia ETB. I neutrofili partecipano alle primissime fasi della risposta infiammatoria nella SSC e contribuiscono al danno endoteliale, alla produzione di specie reattive dell'ossigeno, all\u2019attivazione dei fibroblasti e al reclutamento dei linfociti B e T. Inoltre i neutrofili, sotto stimolo dell\u2019ET-1 sono incoraggiati a produrre citochine proinfiammatorie, come IL-8. IL-17 e TNF. L\u2019ET-1, attraverso l'ETA e l\u2019ETB, pu\uf2 contribuire dunque a innescare l'attivazione dei neutrofili, che contribuiscono al danno vascolare . Considerando che l'espressione ETB \ue8 inferiore nei pazienti affetti dalla forma limitata si SSc piuttosto in quelli con forma diffusa di malattia, l\u2019ETA sembrava essere importante per gli effetti profibrotici innescati dell\u2019ET-1. Poich\ue9 la minor espressione di ETB sui monociti correla con la presenza di ipertensione arteriosa polmonare e una diminuita espressione di ETA sulle cellule T correla con la presenza di interstiziopatia polmonare, possiamo ipotizzare che un diverso pattern di espressione recettoriale \ue8 associato a una differente risposta delle cellule T o dei monociti nell'induzione dell\u2019ipertensione arteriosa polmonare o dell\u2019interstiziopatia polmonare. Pertanto la differente espressione di ETA o ETB pu\uf2 portare allo sviluppo di complicanze cliniche differenti. Considerando che l'espressione ETB \ue8 aumentata sulle cellule T attivate, ETB probabilmente giocato un ruolo importante nel processo infiammatorio. Infine, l\u2019ET-1 ha un\u2019azione modulante sulla risposta immune e il pattern citochinico che i linfociti producono sotto stimolo con ET-1 \ue8 diverso a seconda del recettore dell\u2019ET-1 che \ue8 bloccato. Questi risultati confermano l'ipotesi che ET-1 possa avere un ruolo non solo sulla vasocostrizione e fibrosi ma anche sull\u2019infiammazione .Endothelin-1 (ET-1) plays a pivotal role in vasoconstriction, fibrosis and inflammation, the three major aspects in the pathogenesis of Systemic Sclerosis (SSc). ET-1 receptors are ETA and ETB. The receptors are expressed on the majority of cells involved in SSc, such as endothelial cells, smooth muscle cells and fibroblasts. Little is known about the expression of ET-1 receptors on leukocytes, except for macrophages and monocytes; there is no information about the expression of ETA and ETB on lymphocytes, neutrophils and other cells involved in the innate and acquired immune response. Endothelin receptors antagonists are used in the treatment of scleroderma patients with recurrent ischemic digital ulcers and/or pulmonary arterial hypertension. They have beneficial effects on vasoconstriction and fibrosis, but less is known about their anti-inflammatory role. We aimed at studying the link between ET-1 and inflammation in SSc. Since T and B cells, monocytes and neutrophils are among the most important cells in inflammatory responses in SSc, we studied ETA and ETB expression on these cells with flow cytometry, and also ETA- and ETB-coding mRNA expression in T CD4+ cells and neutrophils by RT-PCR. We have studied the different expression of receptors on T and B cell and monocytes between patients and controls, the correlations of the expression with the cutaneous form of disease, with Bosentan therapy, and ischemic digital ulcers, pulmonary arterial hypertension and interstitial lung disease. We also studied the receptors modulation on activated T cells by flow cytometry. In order to evaluate the pro-inflammatory effects of ET-1 and the anti-inflammatory role of Bosentan, we studied how ET-1 influences IFN-\u3b3 and IL-4 production by T CD4+ cells, with or without receptors blockage. We also studied the expression of IFN-\u3b3-, IL-4- IL-6-, IL-10- and IL-17-coding mRNA in T CD4+ cells by Real Time PCR at different times in order to understand the timing of T CD4+ cells response to stimulation with ET-1. We previously described that T cells and monocytes express both ETA and ETB receptors. We have validated our data in a larger cohort of patients and controls. We confirmed that not only T cells and monocytes but also B lymphocytes and neutrophils express ETA and ETB on their surface. Moreover, the expression of ETA was greater than ETB both in patients and controls in T cells and monocytes, while for B cells there was not difference between ETA and ETB expression. Interesting, neutrophils express both ETA and ETB. Neutrophils participate in the early stages of SSc and contribute to endothelial damages, by production of reactive oxygen species, fibroblasts activation and recruitment T and B cells. ET-1, through ETA and ETB, can contribute to trigger neutrophils activation, that lead to vascular damage. Considering that ETB expression was lower in dSSc- rather than lSSc-patients, ETA signal seemed to be important in the cutaneous profibrotic effects of ET-1. Since a lower ETB expression on monocytes correlated with PAH and a lower ETA expression on T cells correlates with ILD, we can hypothesize that a different pattern of receptors expression is associated with a different response of T cells or monocytes in the preferential induction of PAH or ILD. Therefore ETA or ETB signalling may lead to different clinical features. Considering that ETB expression is increased on activated T cells, ETB signal probably played a major role in inflammation. We also show that activation of ETA and ETB receptors plays an immunomodulatory role, since the production of cytokines changes over time in relation to the stimulation by ET-1 in the presence or absence of the selective blockade of one or both receptors. Furthermore these results support the hypothesis that ET-1 system has a role not only on vasoconstriction and fibrosis but also on inflammation

Endothelin Receptors Expressed by Immune Cells Are Involved in Modulation of Inflammation and in Fibrosis: Relevance to the Pathogenesis of Systemic Sclerosis

open13noEndothelin-1 (ET-1) plays a pivotal role in vasoconstriction, fibrosis, and inflammation, the key features of systemic sclerosis (SSc). ET-1 receptors (ETA and ETB) are expressed on endothelial cells, smooth muscle cells, and fibroblasts, but their presence on immune cells has not been deeply investigated so far. Endothelin receptors antagonists such as bosentan have beneficial effects on vasoconstriction and fibrosis, but less is known about their potential anti-inflammatory effects. We studied the expression of ET-1 receptors on immune cells (T and B lymphocytes, monocytes, and neutrophils) and the link between ET-1 and inflammation in patients with SSc. We show here that ET-1 exerts a proinflammatory effect in CD4+ T cells, since it induces an increased IFN-γ production; preincubation with antagonists of both receptors reduces IFN-γ production. Moreover, following ET-1 stimulation, neutrophils produce proinflammatory mediators, thus amplifying the effects of activated CD4+ T cells. Our data indicate that ET-1 system is involved in the pathogenesis of inflammation and fibrosis typical of SSc, through the activation of T lymphocytes and neutrophils and the consequent release of proinflammatory and profibrotic cytokines. These findings suggest that dual ET-1 receptors antagonist therapy, besides its effect on vasculopathy, has a profound impact on the immune system favouring antiinflammatory and antifibrogenic effects.openElisa, Tinazzi; Antonio, Puccetti; Giuseppe, Patuzzo; Alessandro, Barbieri; Giuseppe, Argentino; Federico, Confente; Marzia, Dolcino; Ruggero, Beri; Giacomo, Marchi; Andrea, Ottria; Daniela, Righetti; Mariaelisa, Rampudda; Claudio, LunardiTinazzi, Elisa; Puccetti, Antonio; Patuzzo, Giuseppe; Barbieri, Alessandro; Argentino, Giuseppe; Confente, Federico; Dolcino, Marzia; Beri, Ruggero; Marchi, Giacomo; Ottria, Andrea; Righetti, Daniela; Rampudda, Mariaelisa; Lunardi, Claudi

Immunophenotypic Analysis of B Lymphocytes in Patients with Common Variable Immunodeficiency: Identification of CD23 as a Useful Marker in the Definition of the Disease

Common variable immunodeficiency (CVID) is a primary immunodeficiency characterized by the failure of B lymphocytes differentiation leading to deficient immunoglobulins secretion. The identified genetic defects account only for a minority of cases. The importance of B cells immunophenotyping in the classification of CVID is known. This procedure can identify alterations on the cell surface molecules expression that could explain some immunological disorders characteristic of CVID. Moreover, some immunophenotypical aspects can correlate with clinical features of the disease. We used this procedure to analyze a cohort of 23 patients affected by CVID, in order to identify the novel alterations of B cells and to find the possible correlations with clinical features. Circulating B cells were studied by flow cytometry incubating whole blood with specific antibodies for B cell surface molecules including CD27, IgM, IgD, CD21, and CD23. We compared the population of “switched memory” IgD− CD27+ B lymphocytes with the population of “switched memory” IgM− IgD− CD23− CD27+ B cells. These last B cells were reduced in patients compared to healthy controls; moreover, IgM− IgD− CD23− CD27+ B cells were lower than IgD− CD27+ B cells in patients with CVID. The reduction of this subset of B lymphocytes correlates more tightly than IgD− CD27+ B cells with lymphadenopathy and airways infections. In conclusion, our findings may help in better identifying patients with CVID

Reconstruction and functional analysis of altered molecular pathways in human atherosclerotic arteries

<p>Abstract</p> <p>Background</p> <p>Atherosclerosis affects aorta, coronary, carotid, and iliac arteries most frequently than any other body vessel. There may be common molecular pathways sustaining this process. Plaque presence and diffusion is revealed by circulating factors that can mediate systemic reaction leading to plaque rupture and thrombosis.</p> <p>Results</p> <p>We used DNA microarrays and meta-analysis to study how the presence of calcified plaque modifies human coronary and carotid gene expression. We identified a series of potential human atherogenic genes that are integrated in functional networks involved in atherosclerosis. Caveolae and JAK/STAT pathways, and S100A9/S100A8 interacting proteins are certainly involved in the development of vascular disease. We found that the system of caveolae is directly connected with genes that respond to hormone receptors, and indirectly with the apoptosis pathway.</p> <p>Cytokines, chemokines and growth factors released in the blood flux were investigated in parallel. High levels of RANTES, IL-1ra, MIP-1alpha, MIP-1beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-17, PDGF-BB, VEGF and IFN-gamma were found in plasma of atherosclerotic patients and might also be integrated in the molecular networks underlying atherosclerotic modifications of these vessels.</p> <p>Conclusion</p> <p>The pattern of cytokine and S100A9/S100A8 up-regulation characterizes atherosclerosis as a proinflammatory disorder. Activation of the JAK/STAT pathway is confirmed by the up-regulation of IL-6, STAT1, ISGF3G and IL10RA genes in coronary and carotid plaques. The functional network constructed in our research is an evidence of the central role of STAT protein and the caveolae system to contribute to preserve the plaque. Moreover, Cav-1 is involved in SMC differentiation and dyslipidemia confirming the importance of lipid homeostasis in the atherosclerotic phenotype.</p

The Multidisciplinary Management of Cutaneous Squamous Cell Carcinoma: A Comprehensive Review and Clinical Recommendations by a Panel of Experts

Simple Summary Cutaneous squamous cell carcinoma is one of the most common forms of cancer. Although most cases are cured with surgical excision, a few tumors are associated with a high risk of local or distant relapse; therefore, it is relevant to identify high-risk lesions among all other low-risk CSCCs for the proper diagnostic and therapeutic management. Chemotherapy achieves mostly short-lived responses that do not lead to a curative effect and are associated with severe toxicities. Recently, PD-1 inhibitor cemiplimab was approved by the regulatory authorities for the treatment of advanced cutaneous squamous cell carcinoma; subsequently, the anti-PD-1 agent pembrolizumab received the approval by the FDA only in the same setting. Here, we provide a literature review and clinical recommendations by a panel of experts regarding the diagnosis, treatment, and follow-up of cutaneous squamous cell carcinoma. Cutaneous squamous cell carcinomas (CSCC) account for about 20% of all keratinocyte carcinomas, which are the most common form of cancer. Heterogeneity of treatments and low mortality are a challenge in obtaining accurate incidence data and consistent registration in cancer registries. Indeed, CSCC mostly presents as an indolent, low-risk lesion, with five-year cure rates greater than 90% after surgical excision, and only few tumors are associated with a high-risk of local or distant relapse; therefore, it is particularly relevant to identify high-risk lesions among all other low-risk CSCCs for the proper diagnostic and therapeutic management. Chemotherapy achieves mostly short-lived responses that do not lead to a curative effect and are associated with severe toxicities. Due to an etiopathogenesis largely relying on chronic UV radiation exposure, CSCC is among the tumors with the highest rate of somatic mutations, which are associated with increased response rates to immunotherapy. Thanks to such strong pre-clinical rationale, clinical trials led to the approval of anti-PD-1 cemiplimab by the FDA (Food and Drug Administration) and EMA (European Medicines Agency), and anti-PD-1 pembrolizumab by the FDA only. Here, we provide a literature review and clinical recommendations by a panel of experts regarding the diagnosis, treatment, and follow-up of CSCC

MIOPATIE INFIAMMATORIE

Le miopatie infiammatorie, note anche come miositi, rappresentano un gruppo di malattie infiammatorie croniche a eziologia sconosciuta,che coinvolgono la muscolatura striata. Le miositi si caratterizzano, dal punto di vista clinico, per la presenza di debolezza e facile esauribilit\ue0 muscolare e, dal punto di vista istologico, per un infiltrato infiammatorio a livello del tessuto muscolare

Autoimmunity and infection in common variable immunodeficiency (CVID)

Common variable immunodeficiency (CVID) is a heterogeneous group of diseases, characterized by primary hypogammaglobulinemia. B and T cell abnormalities have been described in CVID. Typical clinical features of CVID are recurrent airway infections; lymphoproliferative, autoinflammatory, or neoplastic disorders; and autoimmune diseases among which autoimmune thrombocytopenia (ITP) is the most common. The coexistence of immunodeficiency and autoimmunity appears paradoxical, since one represents a hypoimmune state and the other a hyperimmune state. Considering both innate and adaptive immune response abnormalities in CVID, it is easier to understand the mechanisms that lead to a breakdown of self-tolerance. CD21lowB cells derive from mature B cells that have undergone chronic immune stimulation; they are increased in CVID patients. The expansion of CD21lowB cells is also observed in certain autoimmune diseases. We have studied CD21lowB cells in patients with CVID, CVID, and ITP and with ITP only. We observed a statistically significant increase in the CD21lowpopulation in the three pathological groups. Moreover, we found statistical differences between the two groups of CVID patients: patients with ITP had a higher percentage of CD21lowcells. Our data suggest that CD21lowcells are related to autoimmunity and may represent a link between infection and autoimmunity

Crossreactive autoantibodies directed against cutaneous and joint antigens are present in psoriatic arthritis

Psoriatic arthritis (PsA) is a chronic inflammatory disease of unknown origin, characterized by erosions and new bone formation. Diagnosis of PsA is mainly clinical and there are no biomarkers available. Moreover in PsA autoantibodies have not been described so far. Indeed an autoimmune origin has been suggested but never proven. Aim of the study was to investigate the possible presence of autoantibodies typically associated with PsA.We used pooled IgG immunoglobulins derived from 30 patients with PsA to screen a random peptide library in order to identify disease relevant autoantigen peptides.Among the selected peptides, one was recognised by nearly all the patients' sera. The identified peptide (PsA peptide: TNRRGRGSPGAL) shows sequence similarities with skin autoantigens, such as fibrillin 3, a constituent of actin microfibrils, desmocollin 3, a constituent of the desmosomes and keratin 78, a component of epithelial cytoskeleton. Interestingly the PsA peptide shares homology with the nebulin-related anchoring protein (N-RAP), a protein localized in the enthesis (point of insertion of a tendon or ligament to the bone), which represents the first affected site during early PsA. Antibodies affinity purified against the PsA peptide recognize fibrillin, desmocollin, keratin and N-RAP. Moreover antibodies directed against the PsA peptide are detectable in 85% of PsA patients. Such antibodies are not present in healthy donors and are present in 13/100 patients with seroposive rheumatoid arthritis (RA). In seronegative RA these antibodies are detectable only in 3/100 patients.Our results indicate that PsA is characterized by the presence of serum autoantibodies crossreacting with an epitope shared by skin and joint antigens

Modulation of adaptive immune response following intravenous immunoglobulin therapy in common variable immunodeficiency

We describe here the effects of IVIG on gene modulation and on immune system in patients with CVI

Ipertensione portale secondaria a fistola artero-venosa splenica. Descrizione di un caso clinico e revisione della letteratura

Le fistole artero-venose spleniche (FAVS) rappresentano una causa rara ma potenzialmente curabile di ipertensione portale pre-epatica. Circa 100 casi sono stati finora riportati in letteratura. Gli Autori descrivono il caso di un uomo di 46 anni, portatore di aneurisma dell'arteria splenica e di FAVS ad alta portata, trattati mediante legatura dei vasi splenici e splenectomia. Attraverso una revisione della letteratura vengono discussi eziologia, localizzazioni anatomiche, sintomi di presentazione ed aspetti diagnostici e terapeutici di questa rara patologia