ACR Meeting Abstracts

ACR Meeting Abstracts

Abstract Number: 2788

DNA Microarray Analysis of Labial Salivary Glands in Patients with Sjögren’s Syndrome: Comparison with IgG4-Related Disease

Hiroyuki Takahashi1, Hiroto Tsuboi1, Mana Iizuka1, Hiromitsu Asashima1, Tomoya Hirota1, Yuya Kondo1, Isao Matsumoto1, Takayuki Sumida1, Seiji Nakamura2, Sachiko Furukawa2, Masafumi Moriyama3, Yuji Nakai4, Keiko Abe4 and Toshio Yoshihara5, 1Department of Internal Medicine, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan, 2Faculty of Dental Science, Kyushu University, Fukuoka, Japan, 3Section of Oral and Maxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, Fukuoka, Japan, 4Functional Food Science and Nutrigenomics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan, 5Department of Otolaryngology, Tokyo Women's Medical University, Tokyo, Japan

Meeting: ACR/ARHP Annual Meeting 2015

Date of first publication: September 29, 2015

Keywords:

Session Information

Date: Tuesday, November 10, 2015

Title: Sjögren's Syndrome: Translational Insights into Sjögren's Syndrome

Session Type: ACR Poster Session C

Session Time: 9:00AM-11:00AM

Background/Purpose: The previous reports showed that DNA microarray in Sjögren’s syndrome (SS) identified the genes associated with mononuclear infiltrates such as immunoglobulin, human leukocyte antigen, T cell receptor and interferon-inducible genes. However, it is possible that the results might reflect nonspecific gene expression due to inflammatory cell infiltration, since the controls were healthy individuals. Therefore, we conducted DNA microarray analysis in labial salivary glands (LSGs) of SS patients and those of IgG4-related disease (IgG4-RD) patients, which also show inflammation, to compare gene expression in LSGs of SS patients with IgG4-RD patients and to identify genes specifically involved in the pathogenesis of SS.

Methods:

1)     Gene expression was analyzed by DNA microarray in LSGs of SS patients (n=5), IgG4-RD patients (n=5) and healthy controls (HC) (n=3). All patients with SS and IgG4-RD fulfilled the Japanese Ministry of Health criteria for the diagnosis of SS (1999) and the comprehensive diagnostic criteria for IgG4-RD (2011), respectively. Moreover, all of the patients had not received steroids or any immunosuppressive agents. After the obtained microarray data were normalized by FARMS algorithm, Differentially expressed genes (DEGs) upregulated in SS than in IgG4-RD were identified in pairwise comparisons (false discovery rate<0.05) by rank products method. Approval for this study was obtained from the local ethics committee and a signed informed consent was obtained from each subject.

2)     Validation of the results was performed by quantitative PCR using LSGs obtained from other patients with SS (n=11), IgG4-RD (n=11), and HC (n=3) than those examined by DNA microarray.

3)     The protein production of validated genes and expressing cells in LSGs from SS patients and IgG4-RD patients were examined by immunofluorescence assay.

4)     Functional analysis of the DEG was performed using CD4+ T cells isolated from peripheral blood mononuclear cells (PBMCs) by comparing the gene expression of the DEG in peripheral CD4+ T cells and the population of Th17 cells differentiated from peripheral CD4+ T cells in Th17 polarizing conditions between patients with SS and HC.

Results:

1)     In patients with SS, 1785 up-regulated probe sets (corresponding to 1320 up-regulated genes) were identified as DEGs in comparison with those with IgG4-RD.

2)     CXCL9, NR4A2, DPP4, SGK1, and PDK1 were selected as candidate genes for validation, according to rank<150, high expression levels, small variance and relation to T cell functions. PCR validated significantly higher expression of NR4A2 and DPP4 in SS patients than in IgG4-RD patients.

3)     Immunofluorescence staining in LSGs revealed higher production of NR4A2 and DPP4 in SS patients than in IgG4-RD patients and localization of NR4A2 in IL-17+ CD4+ T cells of SS patients.

4)     Peripheral CD4+ T cells of patients with SS showed significantly higher expression of NR4A2, and more preferably differentiated into Th17 cells than those of HC.

Conclusion:

The gene expression pattern of LSG in patients with SS was different from that in those with IgG4-RD.

NR4A2 might be a novel molecule involved in the pathogenesis of SS via Th17 differentiation.

Disclosure: H. Takahashi, None; H. Tsuboi, None; M. Iizuka, None; H. Asashima, None; T. Hirota, None; Y. Kondo, None; I. Matsumoto, None; T. Sumida, None; S. Nakamura, None; S. Furukawa, None; M. Moriyama, None; Y. Nakai, None; K. Abe, None; T. Yoshihara, None.

To cite this abstract in AMA style:

Takahashi H, Tsuboi H, Iizuka M, Asashima H, Hirota T, Kondo Y, Matsumoto I, Sumida T, Nakamura S, Furukawa S, Moriyama M, Nakai Y, Abe K, Yoshihara T. DNA Microarray Analysis of Labial Salivary Glands in Patients with Sjögren’s Syndrome: Comparison with IgG4-Related Disease [abstract]. Arthritis Rheumatol. 2015; 67 (suppl 10). https://acrabstracts.org/abstract/dna-microarray-analysis-of-labial-salivary-glands-in-patients-with-sjgrens-syndrome-comparison-with-igg4-related-disease/. Accessed .

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