Statistical analyses were performed using GraphPad Prism, version 5

Statistical analyses were performed using GraphPad Prism, version 5.0 software (GraphPad Software, San Diego, CA, USA). severity. Fittingly, anti-ER antibody levels were also significantly associated with alterations of immunological features of SSc patients, including increased T cell apoptotic susceptibility and changes in T regulatory cells (Treg) homeostasis. In particular, the percentage of activated Treg Rabbit Polyclonal to CRMP-2 (CD4+CD45RA? FoxP3brightCD25bright) was significantly higher in anti-ER antibody positive patients than in anti-ER antibody negative patients. Taken together our data clearly indicate that anti-ER antibodies, probably the involvement of membrane-associated ER, can represent: i) promising markers for SSc progression but, also, ii) functional modulators of the SSc patients immune system. Introduction Estrogens are well-known regulators of the immune responses and several lines of evidence support a key role for them in the development or progression of numerous diseases, including autoimmune disorders Neohesperidin dihydrochalcone (Nhdc) [1]C[5]. Estrogens, in particular 17-estradiol, directly modulate the function of immune cells by transcriptional activity of nuclear estrogen receptors (ER), i.e., ER and ER [1]. Recently, the expression of functional membrane-associated ER in different cell types including human lymphocytes has been suggested [6]C[8] and autoantibodies specific to ER have been detected in sera from patients with systemic lupus erythematosus (SLE) [9]. These anti-ER antibodies behave as true estrogen agonist and are able to induce cell activation and apoptotic cell death in resting lymphocytes as well as proliferation of anti-CD3 activated T cells. Interestingly, a significant association between anti-ER antibody titer and disease activity was demonstrated [9]. Similarly to SLE, systemic sclerosis (SSc) is an autoimmune disease characterized by multiorgan involvement and circulating autoantibodies against intracellular antigens [10]. The pathogenesis of SSc is complex and incompletely understood. Immune activation, vascular damage, and connective tissue fibrosis are all known to be important in the development of this disease [11], [12]. Overall, a substantial female predominance exists in SSc, with a female-to-male ratio ranging from 31 to 141 [10], suggesting that female sex hormones such as estrogen may play a role in disease Neohesperidin dihydrochalcone (Nhdc) pathogenesis. Nevertheless, only limited information is currently available on the role of estrogens in SSc [13]C[15] and the presence of anti-ER antibodies has not been explored yet. Therefore, the aim of this study was to evaluate anti-ER serum immunoreactivity in patients with SSc and to assess the possible relationship between the presence of anti-ER antibodies Neohesperidin dihydrochalcone (Nhdc) and the clinical and immunological features of the disease. Materials and Methods Ethics Statement This study has been conducted according to the principles expressed in the Declaration of Helsinki. Written informed consent was obtained from all patients and controls, and the study was approved by the Ethical Committee of Policlinico Umberto I, Rome, Italy. Patients and Biological Samples We analyzed sera from 71 consecutive patients with SSc ( Table 1 ). All patients fulfilled the preliminary criteria for SSc as defined by the American College of Rheumatology [16]. SSc was diffuse (dcSSc) in 29 patients and limited (lcSSc) in 42 patients. Disease activity was evaluated using the European Scleroderma Study Group (EScSG) activity index [17]. All 71 patients had anti-nuclear antibody (ANA) (indirect immunofluorescence on Hep-2000 cells), 24 out of 29 dcSSc Neohesperidin dihydrochalcone (Nhdc) patients had anti-topoisomerase I antibodies (anti-Scl70 antibodies) and 17 out of 41 lcSSc patients had anti-centromere (ACA) antibodies (Innogenetics, Gent, Belgium). All patients underwent nailfold capillaroscopy and were divided in three different capillaroscopic patterns: early, active and late [18]. Carbon monoxide diffusion capacity (DLCO) was measured by the single breath method, according to the American Thoracic Society standards [19]. Exclusion criteria were previous or concomitant treatments with immunosuppressive drugs. Twenty-two (31%) patients were on low dose steroids (below 10 mg of prednisone a day) at the time of inclusion in the study. The control group consisted of 90 healthy donors matched for age and sex with the SSc group. For flow cytometry analysis, 34 out of 90 healthy donors were randomly selected as representative of the whole series. Table 1 Demographic and clinical characteristics of SSc patients (n?=?71). Age, median (range) years56 (21C79)Sex, n. of men/n. of women11/60Disease duration, median (range) years8 (1C37)Disease type (dcSSc/lcSSc)29/42EScSG, mean (SD)2 (1.6)DLCO, mean (SD), % of predicted value73.2 (19.7)NC pattern:Early18 (25)Active18 (25)Late35 (49)ACA19 (27)Scl7030 (42)Steroid treatment* 22 (31) Open in a separate window Except where indicated otherwise, values are the absolute number.