Poster Session: Session 1: Gastrointestinal autoimmunity

Vasculitis can cause local or diffuse pathologic changes in the gastrointestinal tract, resulting in nonspesific paralytic ileus, mesenteric ischemia, submucosal edema and hemorrhage, or bowel perforation or stricture. Sytemic vasculitis is known to affect the gastrointestinal tract but the nature of the complication is poorly charaterized.

We reported a 48 year old man came with multiple ulcer in mouth and right leg. He also felt fever, erythema in his left eye, epigastric pain, black ter like feces, and decrease his body weight. He did not complaint about edem in his leg or others place. From physical examination revealed redness in his left eye with loss his sight, muliple ulcer in buccal, epigastric pain, ulcer in his leg with 3-4 cm in diameter with no pus. From the laboratoy results showed normocytic normochromic anemia, leukocytosis with netrophilia, increased D-dimer, hypoalbuminemia, ANA positive with speckled pattern; complement C3/C4 was 76,4/140. Skin biopsy of leg ulcer showed vasculitis. Gastroscopy result showed erosive gastritis and colonoscopy result showed multiple ulcer in colon. Result of biopsy of gastric showed the presence of vasculitis which patohological anatomic result revealed signs of erythrocyte extravacation. He was given methyl prednisolon at immunosuppresant dose, oral anticoagulant with prophylactic dose, proton pump inhibitor and acyclovir. The ulcers were resolved after one month follow up.

A 46 year old female presented to our hospital, with complaints of easy fatiguability and dysphagia of one year duration. Clinical examination revealed pallor and platynychia.

Her investigations showed haemoglobin of 5.6gm%(11-15 gm%), Serum Ferritin - 1.55 ng/ml and peripheral smear - microcytic, hypochromic anemia. Barium swallow (Figure 1) and UGI scopy was done which showed a mucosal web causing luminal compromise.Endoscopic dilation of the web was done. She was transfused with 2 units of packed cells.Iron supplements were given.
A diagnosis of Plummer Vinson syndrome should be considered in any person presenting with iron deficiency anemia and dysphagia because prognosis of Plummer-Vinson syndrome is excellent and it is a reversible cause of dysphagia.
P1-035 CALPROTECTIN AND ANCA IN NEUTROPHIL-MEDIATED IBD
I. Lochman¹, A. Lochmanová², J. Martinek³, H. Tomaskova⁴, I. Bártková⁵, P. Svoboda⁶, V. Smajstrla<sup>7


1</sup>Immunology, Spadia Lab Ltd, Ostrava, Czech Republic
²Department of Biomedical Sciences, Faculty of Medicine University of Ostrava, Ostrava, Czech Republic
³Immunology and Allergy, Institute of Public Health, Ostrava, Czech Republic
⁴Department of epidemiology and Public Health, Faculty of Medicine University of Ostrava, Ostrava, Czech Republic
⁵5Department of Pediatrics, Municipal Hospital, Ostrava
⁶6Department of Clinical Studies, Faculty of Medicine, University of Ostrava
⁷7Bormed, Ltd., Ostrava, Czech Republic

Determination of faecal calprotectin and ANCA are used in the diagnosis and laboratory monitoring of inflammatory bowel diseases - IBD (Crohn's disease, ulcerative colitis and neoplasms). Both analytes are associated with an inflammatory process which involves neutrophils, but the information obtained from these tests is not completely identical.

We analyzed the results of faecal calprotectin using Quantum blue (Buehlmann) and ELISA (R-Biopharm) kits and ANCA determination using IFA CIBD-Profile 1 kits (Euroimmun) containing chips with ethanol-fixed granulocytes and IgG conjugate in our labs during 2011 and 2012. Overall, 1088 samples from 823 patients have been analyzed for calprotectin and ANCA and 4723 samples only for ANCA using IFA CIBD-Profile 1 kits.

Detailed analysis of results confirmed that the levels of both analytes correlate with the intensity of neutrophil-mediated inflammation. We demonstrate it on several case reports. The vast majority of total ANCA was designated as A-ANCA, possibly GS-ANA after typing, but it was difficult or impossible to distinguish these antibodies in the presence of other antinuclear antibodies in some cases.

However targets against which these antibodies are formed, are mostly NETs epitopes (neutrophil extracellular traps) that is difficult, if not impossible, to define generally. Eventhough IFA on ethanol-fixed granulocytes is generally recommended for ANCA screening, it has not been uniquely accepted legal diagnostic scheme for ANCA screening and their subsequent typing. Especially for diseases other than ANCA associated vasculitis (AAV) can be obtained frequently confused results between IFA and other immunoassays that use defined neutrophil cytoplasmic substrates.
P1-036 THE TETRASPANIN CD151 AS A NOVEL REGULATOR OF T CELL MOTILITY IN HEALTH AND INFLAMMATORY BOWEL DISEASE
E. Toister-Zelman¹, E. Bakos¹, S. Cohen¹, E.H.U.D. Zigmond¹, V. Grabovsky¹, A.D.I. Sagiv¹, G.I.L.I. Hart¹, N. Kaushansky¹, A.V.I. Ben-Nun¹, A. Sonnenberg¹, R. Alon¹, I.D.I.T. Shachar<sup>1


1</sup>Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel

The continuous recirculation of mature lymphocytes and their entry into the peripheral

lymph nodes (LNs) are crucial for the development of an effective immune response.

Picomolar (pM) levels of CCL2 were previously shown to inhibit T cell migration to LNs

resulting in reduced inflammation. However, the molecular mechanisms regulated by low

dose CCL2 remained unknown. In this study, we demonstrate that the tetraspanin CD151 is a target

of CCL2 in T cells. pM levels of CCL2 downregulated CD151 expression and dissociated

CD151/VLA-4 and CD151/LFA-1 complexes in T lymphocytes, resulting in their attenuated

migration. Ablation of CD151 expressed on immune cells reduced inflammation in a DSS

induced colitis model of CD151-/- chimeric mice. Moreover, blocking of CD151, using CD151

truncated peptide fragment, reduced cell migration in vitro and in vivo, and inhibited the

development of DSS-induced colitis. Thus, CD151 is a novel orchestrator of T cell motility,

Several independent genome wide association studies have shown associations of single nucleotide polymorphisms within the C-type lectin domain family 16 member A gene, CLEC16A, with various autoimmune diseases, including systemic lupus erythematosus (SLE). However, the expression and function of this novel C-type lectin domain-containing protein is still largely unknown. This study aimed to relate its expression level in peripheral blood mononuclear cells (PBMCs) with SLE disease activity. Two expressed isoforms, isoforms 1 and 2, of CLEC16A were cloned and sequenced from PBMCs from healthy Chinese. Surprisingly, both sequences of isoforms 1 and 2 bore differences when compared to the reference sequences derived from Caucasians. In isoform 1, exon 5 was consistently absent in Chinese PBMCs. However, in isoform 2, the stretch of 48-bp in-frame deletion within exon 11 in the reference sequence was found present in Chinese PBMCs. The mRNA expression levels of both isoforms were found significantly lower in SLE patients than in normal individuals. Yet, their expression levels did not correlate with disease activity as measured by SLE disease activity index. The molecular and cellular functions of CLEC16A are currently under investigation, which should shed light on the importance of this protein in SLE pathogenesis.

Background: The insulin-like growth factor (IGF) system plays a prominent role in the regulation of immunity and inflammation. Inappropriate balance of the IGF-1 signalling has been reported in autoimmune disorders. A few previous studies have investigated the modulation of the IGF signalling pathway in the rheumatic diseases, but their findings are controversial. Recently, single nucleotide polymorphism (SNP) located in exon 16 (+3179G/A; rs2229765) of IGF-1R gene was found associated with serum levels of IGF-I and human longevity.

Aim: To compare +3179G/A IGF-1R genotype distribution in rheumatoid arthritis (RA) and ankylosing spondylitis (AS).

Patients and methods: Genotyping for +3179G/A polymorphism was performed by restriction fragment length polymorphisms (RFLP)-PCR assay in 70 patients with RA, 55 with AS and corresponding age and sex matched healthy donors.

Results: We found an opposite effect of the investigated IGF-1R polymorphism (+3179G/A; rs2229765) on disease susceptibility to RA and AS. Significant differences in allele and genotype frequencies between RA and healthy controls were observed: higher frequency of homozygous genotype AA (22.9% vs14.1%; OR= 2.33, p=0.034) and allele *ÒAÓ (47.9% vs37.5%; OR= 1.53, p=0.032) in cases versus controls. Although the genotype frequencies in AS in comparison with healthy controls were not significantly differing, the lowest frequency of AA genotype was observed in AS (10.9% vs 20,9%; OR= 0,47; p=0.142).

Conclusions: Allele A and genotype AA of +3179G/A IGF-1R polymorphism are associated with RA genetic predisposition, but it seems as a protective factor for AS. Our data confirm the involvement of the IGF-1 signal pathway in RA.

The shared pathogenesis between autoimmune diseases has been explored through comorbidity studies. Genome-wide association studies (GWAS), which compare genetic data between cases and controls, provide support of many genetic risk factors shared between diseases. Studies have specifically explored this shared pathogenesis via GWAS summary statistics alone; such as testing the correlation of association signals between pairs of autoimmune diseases across many SNPs. Here we introduce a new method that rigorously explores the shared genetic risk between autoimmune diseases and between them and diseases of other classes. Our method applies principal component analysis (PCA) to a matrix of gene-level significance scores across many GWAS datasets, while correcting for confounders such as genotyping arrays and sample size. We applied our method to 30 datasets, of which 14 are autoimmune diseases. Other disease categories include psychiatric, neurological and cancers. We report several findings: 1) different studies of the same diseases generally lie closer together. 2) Inflammatory bowel diseases cluster together away from other autoimmune diseases. 3) Some psychiatric disorders cluster with autoimmune diseases. 4) Genes that underlie the observed structure are significantly enriched for immune related pathways such as IL1-mediated signaling. Prompted by our findings, we carried out a GWAS with combined cases of bipolar and type-1 diabetes (T1D). We found an association near the gene PTPRM, which presents a new association to T1D. In summary, our method offers novel findings regarding the shared pathogenetics of autoimmune diseases.

Background and aims: Previous genome-wide association study by WTCCC identified rheumatoid arthritis (RA) susceptibility loci in British. Here, we recruited 1894 subjects to investigate whether the three SNPs with strong RA association signal in British was associated with RA in Han Chinese. Methods: We analyzed the three SNPs in Han Chinese population based on the HCB data of HapMap. The rs6679677 does not exist in Han Chinese. Other two SNPs (rs6457617 and rs11761231) were performed case-control study in the test cohort of 380 subjects firstly and the positive result was further replicated in another validation cohort of 1514 subjects. Results: In the test cohort, only rs6457617 was significantly associated with RA. The individuals bearing the homozygous genotype CC had 0.39-fold risk than these bearing the wild-type genotype TT (P = 0.004, OR 0.39, [95%CI 0.21-0.74]). And the protective effect of allele C was confirmed in combined analysis with test and validation cohort(P_{genotye CC/TT} = 9.4*10^-12, OR 0.35, [95% CI 0.26-0.47], Table). Conculsions: The rs6457617, which locates between the HLA-DQB1 and HLA-DQA2 encoding β chain of MHC-II protein, was also associated with RA in Han Chinese.