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PReS-FINAL-2020: Cell type specific transcriptome analysis in patients with enthesitis related arthritis category of juvenile idiopathic arthritis (JIA-ERA)

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Pediatric Rheumatology201311(Suppl 2):P33

https://doi.org/10.1186/1546-0096-11-S2-P33

Published: 5 December 2013

Keywords

  • Arthritis
  • Peripheral Blood Mononuclear Cell
  • Gene Expression Profile
  • Synovial Fluid
  • Juvenile Idiopathic Arthritis

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 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 signaling and leukocyte migration.Results obtained with monocytes are summarized below in table 1.

Table 1

Groups compared

Genes up regulated

Genes down regulated

Number of dysregulated pathways [total (significant)]

Pathways of immunological relevance

EB vs EF

776

189

19 (12)

Cell adhesion, IgA production, antigen processing, lysosomal processing

CBMO vs EBMO

821

1251

21 (12)

Cytokine signaling, TLR signaling, antigen presentation, chemokines signaling

EBMO vs EFMO

595

512

17 (9)

Complement cascade, cytokine signaling, antigen 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.

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.

Authors’ Affiliations

(1)
Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical sciences, Lucknow, India

Copyright

© Aggarwal et al.; licensee BioMed Central Ltd. 2013

This article is published under license to 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.

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