BACKGROUNDUrate through Nacht Domain, Leucine-Rich Repeat, and pyrin domain-containing protein 3 (NALP3) dependent caspase-1 activation stimulates macrophages to secrete inteleukin-1β (IL-1β). Purinergic receptor P2X7 plays a role in the urate induced NALP3 activation. Urate also enhances adaptive immunity indirectly through its effect on antigen presenting cells. In this study, the direct effect of urate on primary human lymphocytes was evaluated.METHODSLymphocytes were cultured with or without monosodium urate crystals in the presence or not of a P2X7 inhibitor. Caspase-1 activity was assessed colorimetrically in cell lysates and IL-1β was measured in supernatants with ELISA. Whole lymphocyte viability and proliferation, as well as T-cell proliferation were assessed by means of 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay and of flow cytometry respectively.RESULTSUrate induced caspase-1 activation and IL-1β release by lymphocytes. It also induced proliferation of whole lymphocytes and T-cells as well. P2X7 inhibitor abrogated lymphocyte proliferation.CONCLUSIONSUrate, a well defined danger signal, stimulates directly human lymphocytes in a P2X7 dependent way. The subsequent IL-1β secretion could enhance inflammation, whereas expansion of lymphocyte clones could facilitate a subsequent adaptive immune response.

printing supported by . Visit Chiesi at Stand B2.10 SUNDAY, SEPTEMBER 2ND 2012 172 Whole-body magnetic resonance imaging in sarcoidosis to assess extrapulmonary organ involvement Katrin Hostettler1, Ueli Studler2, Vlad Bratu2, Arne Fischmann2, Michael Tamm1. 1Clinic of Respiratory Medicine, University Hospital, Basel, BS, Switzerland; 2Department of Radiology, University Hospital, Basel, BS, Switzerland Introduction: Sarcoidosis is a systemic inflammatory disorder that may affect any organ of the body. There is no valid tool to assess the extent of extrapulmonary organ involvement. Whole-body magnetic resonance imaging (MRI) might be a promising modality to detect extrapulmonary disease activity. Aim: To assess the validity of whole-body MRI with regard to extrapulmonary disease activity in patients with sarcoidosis. Methods: 24 consecutive patients at the Clinic of Respiratory Medicine, University Hospital Basel, Switzerland, with histologically-confirmed sarcoidosis were prospectively included. All patients underwent whole-body MRI. Pulmonary function tests and the extrapulmonary physician organ severity tool (ePOST) were assessed for each patient. Results: In total 9/24 (38%) patients showed findings of probable or possible sarcoidal origin. 5/24 (21%) showed skeletal lesions, 3/24 (13%) had muscular findings and 1/24 (4%) had enhancement of the cauda equina. ePOST score was significantly higher in those 9 patients with abnormal whole-body MRI-findings (17.3) than in those with normal images (10.6). FVC percentage predicted (%P), TLC,%P, and DLCO,%P were significantly lower in those patients with abnormal skeletal enhancement compared to those without skeletal abnormalities. Conclusions: Whole-body MRI depicted manifestations of extrapulmonary sarcoidosis in 38% of cases in an unselected patient sample. Abnormal whole-body MRI findings correlated with high ePOST scores, and might thus be a valid tool to assess extrapulmonary disease activity. Abnormal skeletal findings correlated with decreased lung volumes, and might therefore be a marker of total disease activity. 173 HLA-DPB1 and chronic sarcoidosis Annika Wennerström1, Anne Pietinalho2, Jagoda Lasota1, Krista Salli1 , Marja-Liisa Lokki1, Olof Selroos3. 1HLA Laboratory, Haartman Institute, Helsinki, Finland; 2Raasepori Health Care Centre, Raasepori Health Care Centre, Raasepori, Finland; 3Semeco AB, Semeco AB, Vejbystrand, Sweden Background: Beryllium disease and sarcoidosis have clinical similarities. Beryllium exposure together with the HLA-DPB1 alleles containing a glutamic acid at amino acid position 69 (Glu69) is associated with beryllium sensitization and chronic beryllium disease (CBD). Aim: To determinate whether the same genetic variations of HLA-DPB1 that are found in CBD are associated with chronic sarcoidosis. Methods: HLA-DPB1 was determined in 98 Finnish patients with chronic sarcoidosis not resolved within 2 years and in 150 control subjects. The DPB1 alleles were genotyped with sequence specific primers (Olerup SSPTM) and haplotypes were formed with MHC class II and class III markers. Results: 17 different DPB1 alleles were observed. The DPB1*04:02 allele was less frequent among the patients (10.2% vs.20.3%; p=0.003, OR=0.45) than controls. A haplotype with DPB1*04:01, one C4A gene and one C4B gene was increased among sarcoidosis patients (30.9% vs. 22.5%; p=0.036, OR=1.5). By studying polymorphic amino acid residues of DPB1, we did not detect an association of Glu69, but a DPB1*04:02 specific amino acid variant was detected at the position 178 (Met) suggesting a protective role for chronic sarcoidosis (p=0.004, OR=0.42). Furthermore, preliminary SNP analyses of HLA class II and III region showed that the SNP associations are independent from DPB1. Conclusion: We confirm and further describe the contributory role of DPB1 with the risk associated for chronic sarcoidosis. Preliminary results suggest that both DRB1 and certain independent markers in the HLA class II and III region increase the risk for chronic sarcoidosis. However, HLA association analyses are complicated by the extensive linkage disequilibrium (LD) across the HLA region. 174 Evaluation of ANXA11 rs1049550 SNP linkage with sarcoidosis susceptibility António Morais1,2, Bruno Lima3, Sandra Tafulo3, Maria Peixoto3, Helena Alves3, Natalia Melo1, Patricia Mota1, Agostinho Marques1,2, Luis Delagado4,5. 1Pneumology, Centro Hospitalar São João, Oporto, Portugal; 2Pneumology, Faculdade de Medicina do Porto, Oporto, Portugal; 3Genetics, Centro de Histocompatibilidade do Norte, Oporto, Portugal; 4Imunoalergology, Hospital São João, Oporto, Portugal; 5Immunology, Faculdade de Medicina do Porto, Oporto, Portugal Introduction: A recent genome-wide association detected a strong association between Annexin A11 (ANXA11) polymorphisms and sarcoidosis susceptibility. ANXA11 take part in several biological pathways, namely apoptosis and proliferation. Aim: Evaluation of ANXA11 rs1049550 SNP association with sarcoidosis susceptibility in a Portuguese population and after stratification of clinically distinct disease presentations. Methods: A case-control study included 136 unrelated patients (37.7±12.6 years, 58.8% women) and 92 healthy controls (32.3±7.0 years, 62% women). Allele frequencies were compared with Chi-square (or Fisher exact test when appropriate) and genotype frequencies with Chi-square for trend test. Odds ratios (OR) and 95% CIs were calculated as association measures. Samples were genotyped for ANXA11 rs1049550 C/T (R230C) polymorphism using real time PCR with TaqMan SNP genotyping assay. Results: The frequency of ANXA11 rs1049550 (R230C)*T allele was significantly lower in sarcoidosis patients (33.1%) compared with controls (46.2%, p<0.01, OR=0.58, 95%CIs=]0.38;0.86[). An OR=0.44 and OR=0.4 for sarcoidosis was obtained respectively, in the carriers of one (genotype ANXA11 CT) and two (genotype ANXA11 TT) copies of ANXA11 rs1049550*T allele normalised to the CC wild type genotype (p<0.01). When patients with erythema nodosum (EN) were removed, this association persists only in the group without EN. There was no significant difference among radiological Scadding stages. Conclusions: In this population an association between ANXA11 rs1049550*T SNP and protection to sarcoidosis was observed, confirming previous data in populations from different geographic regions, but only in those patients without EN. 175 Bosentan for sarcoidosis associated pulmonary arterial hypertension (BoSAPH) was effective in advanced parenchymal lung disease Robert Baughman1, Daniel Culver2, Francis Cordova3, Maria Padilla4, Kevin Gibson5, Elyse Lower1, Peter Engel6. 1Intern Med, University of Cincinnati, United States; 2Respiratory, Cleveland Clinic, Cleveland, United States; 3Intern Med, Temple University, Philadelphia, United States; 4Intern Med, Mount Sinai, New York, United States; 5Intern Med, University of Pittsburgh, United States; 6Cardiology, Ohio Heart and Cardiovascular Center, Cincinnati,

Peripheral blood from a 10-year old female was referred to the Immunology Lab for immunophenotyping. The patient complained for recurrent upper respiratory tract infections and otitis media during the last year while a history of frequent infections was recorded since infancy. Although, the absolute number of lymphocyte subsets was into normal ranges (fig. 1 and tab. 1), the patient displayed a mild hypogammaglobulinemia (tab. 1). Although the patient did not fulfill the diagnostic criteria for the diagnosis of common variable immunodeficiency (CVID), a molecular analysis of TNFRSF13B gene (encoding TACI) was performed.