JUVENILE IDIOPATHIC ARTHRITIS PATIENTS
Manuela Capone1, Laura Maggi1, Veronica Santarlasci1, Mariacaterina Rossi1, Alessio Mazzoni1, Gianni Montaini1, Rolando Cimaz2, Matteo Ramazzotti3, Giusi Barra4, Marie Pierre Piccinni1, Raffaele De Palma4, Francesco Liotta1, Enrico Maggi1, Francesco Annunziato1, Lorenzo Cosmi1
1Department of Clinical and experimental medicine, University of Firenze, Firenze
2Anna Meyer Children’s Hospital and University of Firenze, Firenze
3Department of Clinical Experimental and Biomedical Science “Mario Serio”, University of Firenze, Firenze
4Dept. of Clinical & Experimental Medicine, Second University of Napoli, Napoli
Purpose: In the present study we aim to explore chitinase 3-like- 1 (CHI3L1) expression in T helper cell subsets as well as to found any possible correlation between its production and T helper cells involvement in chronic inflammatory processes.
Methods: A microarray analysis of gene expression profile was performed on Th17 and classic Th1 cell clones and CHI3L1 was found among the up-regulated genes on Th17 cells. Different types of helper T cell clones (TCCs) were then evaluated by Real Time PCR (RT-PCR) for CHI3L1 mRNA expression; protein expression was investigated in cell lysates by western blotting and in cultures supernatants by ELISA. ELISA was also used to measure CHI3L1 in the serum and in the synovial fluid (SF) of Juvenile Idiopathic Arthritis (JIA) patients.
Results: At mRNA level CHI3L1 was highly expressed by Th17, Th17/Th1, non classic Th1 and even in Th17/Th2 cell clones, whereas it was virtually absent in CD161- classic Th1 and Th2 TCCs. CHI3L1 was also detected in cell culture supernatants of Th17 and Th17-derived cells but not of classic Th1. Moreover CHI3L1 was higher in the SF than in serum of JIA patients, and it positively correlated with the frequency of Th17 and non-classic Th1 cells in SF. CHI3L1 in SF also positively correlated with the C reactive protein (CRP) serum levels, and with the levels of some proinflammatoty cytokines, such as IL-6 and p40, which is the common subunit of IL12 and IL23.
Conclusions and Discussion: Here we describe for the first time CHI3L1 production by T cells owing the Th17 family. Moreover the positive correlation found between the frequency of Th17 and Th17-derived cell subsets and CHI3L1 levels in SF of JIA patients, in agreement with the suggested role of these cells in inflammatory process, candidates CHI3L1 as a possible biological target in JIA treatment.
References
1. Lee CG et al. PMID: 21054166 2. O’ Shea JJ et al. PMID: 18172584
P2.11
RETROSPECTIVE AND PROSPECTIVE LONGITUDINAL EVALUATION OF A COHORT OF PATIENTS AFFECTED BY SYSTEMIC SCLEROSIS COMPLICATED BY PULMONARY ARTERIAL HYPERTENSION
Elisa Tinazzi1, Federico Confente1, giuseppe Patuzzo1, Claudio Lunardi1
1Department of Medicine, University of Verona, Verona
Systemic sclerosis (SSc) is an autoimmune disease characterized by a complex pathogenesis which combine aspects of endothelial dysfunction, immune abnormalities, both at humoral and cellular levels, and diffuse fibrosis.
Pulmonary arterial hypertension (PAH) affects about 7-12% of SSc patients and is the major cause of death.
With the aim to analyze the prevalence of PAH in a cohort of SSc patients and to study any possible interference or relation between PAH and other clinical features or laboratoristic parameters, we enrolled 118 SSc patients for a retrospective/ prospective longitudinal study. We collected SSc patients data from January 2000 to December 2016. The study included 98 females and 20 males; during the period in examination (01/2000 – 12/2016), 18 patients died, 16 females and 2 males. According to LeRoy and Medsger’s criteria patients were classified as affected either of limited or diffuse SSc (respectively lSSc or dSSc). Considering the subset of the disease, 78 patients (65,7%) were affected by lSSc and 40 patients (34,3%) by dSSc. The following data of each patient involved in this study ta were collected: age, sex, cutaneous fibrosis (estimated confronting modified Rodnan Skin Score), positivity of ANA, anti-centRomare antibody and anti-topoisomerase II antibody, and specific clinical manifestations such as presence of digital ulcers (active or past), involvement of digestive system, interstitial lung disease and pulmonary hypertension.
We focalized our attention on the group of patients affected by pulmonary hypertension and analysed the results of all instrumental examinations such as transthoracic rest echocardiographic test, right heart catheterization, high resolution chest CT, pulmonary function tests, 6 minute walking test at time of diagnosis and in the follow-up, particularly after the beginning of either endothelin-1 receptors antagonists (ERAs) or 5 phosphodiesterase inhibitors (5PDE-i).
In particular we considered these patients with the aim of evaluate any clinical feature or laboratoristic items which may have a predictive value for the onset of the clinical manifestation or a prognostic value of the same organ involvement.
Our data showed a significant correlation between the onset of PAH and high levels or rapid increase of ANA title and/or presence of anticentRomaric pattern. Furthermore, the “active” and “late” capillaroscopic pattern as well as the presence of hystory of skin ulceration or active digital ulcers are also associated with the onset of IAP. Our study also confirms that an earlier diagnosis of PAH lead to an early treatment with positive effect on quality of life and/or prognosis.
P2.12
RAI SIGNALING IN ASTROCYTES PLAYS A CRITICAL ROLE IN SHAPING THE CENTRAL NERVOUS SYSTEM MICROENVIRONMENT DURING EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS
Domiziana De Tommaso1, Cristina Ulivieri1, Francesca Finetti1, Giuliana Pellicci2, Mario Milco D’Elios3,
Clara Ballerini4, Cosima Tatiana Baldari5
1Department of Life Sciences, University of Siena, Siena 2Department of Experimental Oncology, European Institute of Oncology, Milano
3Department of Experimental and Clinical Medicine, 4Department of Neurosciences, University of Firenze, Firenze Purpose: Multiple sclerosis (MS) is an autoimmune disease where encephalitogenic Th1/Th17 cells have been long considered responsible of the neuroinflammatory reaction. Recent data indicate however that the local central nervous system (CNS) microenvironment, which is controlled by both infiltrated autoreactive T cells and CNS resident cells, plays a crucial role in disease onset and progression. Among CNS resident cells, astrocytes actively modulate the autoimmune response during MS shaping T cells responses, however, molecular mechanism underlying this function are still poorly defined1. We have recently
identified Rai as a novel astrocytic adaptor responsible for the TrkB- and IL-17R–dependent production of proinflammatory mediators in astrocytes, whose loss in mice ameliorates the experimental autoimmune encephalomyelitis (EAE), in face of higher frequency of CNS infiltrated Th17 cells highlighting the key role played by astrocytes during EAE2.
Methods: To investigate the mechanism through which Rai-/- astrocytes modulate the function of encephalitogenic T cells we have used both culture supernatants from encephalitogenic T cells and co-cultures of Rai+/+ and Rai-/- astrocytes with T cells
to measure the expression of ATP hydrolyzing enzymes CD39 and CD73 on astrocytes as well as the release of cytokines from these cells.
Results and Discussion: Here we show that Rai-/- astrocytes release
higher level of Th17-inhibitory cytokines, IL-10 and IL-27 and upregulate CD39 and CD73 expression which are responsible for the conversion of ATP into the immunosuppressive molecule adenosine more efficiently compared with control astrocytes following exposure to culture supernatants indicating that not only can astrocytes directly affect Th17 cells but generally modulate T cell responses trough contact independent mechanism.
Conclusions: Rai deficiency in astrocytes contributes to establish an unfavourable microenvironment for CNS-infiltrated T cells during EAE.
References
1. Correale and Farez Front. Neurol 2015; 180. 2. Ulivieri et al. J Immunol 2016; 480-90.
P2.13
EFFECT OF BELIMUMAB TREATMENT ON REFRACTORY SYSTEMIC LUPUS ERYTHEMATOSUS: CLINICAL AND BIO- LOGICAL RESPONSE
M. Prete, P. Leone, M.A. Frassanito, V. Desantis, S. Cicco, V. Racanelli, A. Vacca
Department of Biomedical Sciences and Human Oncology. Unit of Internal Medicine, University of Bari Medical School, Bari Purpose: To evaluate the effect of belimumab on clinical, se- rological and immunological parameters of refractory systemic lupus erythematosus (SLE) patients and its association with dis- ease activity.
Methods: 25 active SLE patients, according to the American College of Rheumatology classification, were treated with beli- mumab (10 mg/kg/day, at days 0, 14, 28 and every 28 days). Ten healthy subjects were enrolled as control group. The mean±SD follow-up was 18+-6 months. We also performed flow cytomet- ric analysis of studied T CD4+ cell subpopulations to assess Treg and Th17 subsets at T0, and after six months of treatment (T6). Disease activity was measured with the SELENA-SLEDAI index. Results: In all SLE patients treated with belimumab a signif- icant reduction of SELENA-SLEDAI score (13.66 vs. 5.13, p = 0.012) and of the anti-dsDNA antibody levels (87.69 vs. 11.11 p = 0.028) were observed after six months. An increase of serum C3 (0,75 vs.1,04, p = 0,05) and C4 (0.15 vs. 0.4 p=0,028) and a significant reduction of the prednisone dosage per os (17.08 vs. 5.94, p=0,018) were also obtained. Flow cytometric analy- sis revealed that at T0 patients presented significantly reduced values of CD4+FoxP3+ compared to the controls (0.22±0.12% vs. 0.83±0.2%, p<0.0001) whereas at T6, a significant increase of Tregs was observed reaching values overlapping those of healthy subjects (1.04±0.22). The Th17 lymphocytes’ frequency was sig- nificantly increased in patients with active SLE at T0 (3.5±2.5% vs. 1.4±0.21%, p<0.0002) and reduced at T6 (2±2.2%, p<0.001). Functional in vitro studies showed that the inhibitory capacity of Tregs purified from patients who received Belimumab was simi- lar to that of Tregs purified from healthy subjects.
Conclusion: Belimumab treatment showed a significant reduc- tion of the disease activity in patients with active SLE. A influ- ence of anti-Blys therapy of regulation Treg /Th17 population homeostasis and restoration of their functional state was clearly documented . These changes may represent an pivotal target to inhibit the abnormal self-tolerance in SLE.
P3 - B CELL ITEMS