0Aggarwal et al. Pediatric Rheumatology 2013, 11 (Suppl 2):P33
http://www.ped-rheum.com/content/11/S2/P33 * % PEDIATRIC
RHEUMATOLOGY
POSTER PRESENTATION Open Access
PReS-FINAL-2020: Cell type specific transcriptome analysis in patients with enthesitis related arthritis category of juvenile idiopathic arthritis (JIA-ERA)
A Aggarwal*, A Myles, P Gaur
From 20th Pediatric Rheumatology European Society (PReS) Congress Ljubljana, Slovenia. 25-29 September 2013
Introduction
Enthesitis Related Arthritis Category of Juvenile Idiopathic Arthritis (JIA-ERA) is the most common category of JIA seen in Asian Indians. Transcriptome analysis is a useful tool to analyse pathways involved in disease pathogenesis. Peripheral blood mononuclear cells (PBMC) and SFMC analysis showed involvement of innate immune cells in JIA-ERA. However PBMC/SFMC have variable number of different cells and that can affect interpretation. No data is available on cell type specific transcriptome analysis of blood and synovial fluid in children with JIA-ERA.
Objectives
To study the cell type specific transcriptome analysis of blood and synovial fluid in children with JIA-ERA.
Methods
Six samples each of peripheral blood and synovial fluid were collected from patients with ERA-JIA. Blood from
Table 1
6 healthy controls was also collected. Mononuclear cells were separated by density gradient centrifugation. B cells, T cells and monocytes were separated using MACS columns and purity assessed by flow cytometry. After RNA extraction and checking the quality of RNA (RIN > 8) microarray was done using Illumina chips WG 12 for whole PBMC/SFMC population, T cells, B cells and monocytes. Some of the significant genes were validated by qRT-PCR.
Results
Unsupervised hierarchical clustering revealed that cell subsets could be distinguished based on their gene expression profile. No significant differences were observed between PBMC of patients and healthy controls. Comparison of SFMC and PBMC reconfirmed the results seen earlier. Among T cells and B cells the differential athways identified were related to inflammation like Cell adhesion, antigen processing, cytokine and chemokine signaling,BCR
Groups Genes up Genes down Number of dysregulated pathways Pathways of immunological relevance compared regulated regulated [total (significant)]
EB vs EF 776 189 19 (12) Celladhesion, IgA production, antigen processing,
lysosomalprocessing
CBMO vs 821 1251 21 (12) Cytokine signaling, TLR signaling, antigen
EBMO presentation, chemokines signaling
EBMO vs 595 512 17 (9) Complement cascade, cytokine signaling, antigen
EFMO presentation
EB: ERA blood mononuclear cells, CB: control blood mononuclear cells, EF: ERA Synovial fluid mononuclear cells, Mo: CD14+ monocytes, The major differences were found in monocyte subset. TLR pathway was one of the major pathway identified besides antigen presentation, cytokine and chemokine signalling.
ClinicalImmunology, Sanjay Gandhi Postgraduate Institute of Medical sciences, Lucknow, India
© 2013 Aggarwal et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
BioMed Central
Aggarwal et al. Pediatric Rheumatology 2013, 11 (Suppl 2):P33 Page 2 of 2
http://www.ped-rheum.com/content/11/S2/P33
signaling and leukocyte migration.Results obtained with monocytes are summarized below in table 1.
Conclusion
Monocyte probably play a major role in pathogenesis of JIA-ERA and TLR signalling may be the pathway involved.
Disclosure of interest
None declared.
Published: 5 December 2013
doi:10.1186/1546-0096-11-S2-P33
Cite this article as: Aggarwal et al.: PReS-FINAL-2020: Cell type specific transcriptome analysis in patients with enthesitis related arthritis category of juvenile idiopathic arthritis (JIA-ERA). Pediatric Rheumatology 2013 11 (Suppl 2):P33.
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