A Pesenacker

B Thirugnanabalan

F Warden

H Varsani

J Lewis

L Wedderburn

N Makengo

S De Leeuw

Pediatric Rheumatology

PubMed

Springer - Publisher Connector

POSTER PRESENTATION Open Access
PReS-FINAL-1018: Can the CD4/CD8b ratio be
used as a predictive biomarker in extended-to-be
oligoarticular JIA?
S De Leeuw1,2*, B Thirugnanabalan1, H Varsani1, F Warden1, N Makengo1, J Lewis3, A Pesenacker1, L Wedderburn1
From 20th Pediatric Rheumatology European Society (PReS) Congress
Ljubljana, Slovenia. 25-29 September 2013
Introduction
Predicting disease course in Juvenile Idiopathic Arthritis
(JIA) is difficult. During the first 6 months of oligoarticu-
lar JIA (O-JIA) disease presents with 4 or fewer affected
joints, however after 6 or more months severity might
increase with more than 4 joints involved (extended O-
JIA), or persist with 4 or fewer joints affected (persistent
O-JIA). However patients do not show any clinical differ-
ences at onset making a decision on treatment strategy
difficult before extension occurs. It has previously been
shown that the CD4/CD8 T cell ratio in extended-to-be
patients (samples before extension has occurred) is lower
compared to those who persist (Hunter et al 2010), and
thus might be a useful biomarker in predicting the course
of disease. However measuring this cell ratio currently
requires specialized expertise. To translate the measure-
ment of the CD4/8 synovial T cell ratio as a biomarker a
method is needed, which could be used in a wide range
of hospital facilities, hence a real-time quantitative PCR
(qPCR) method was tested.
Objectives
The aim of this study was to measure CD4 and CD8
mRNA in whole synovial fluid collected at therapeutic
joint injection and to compare it to the gold standard of
flow cytometry using mononuclear cells.
Methods
A total of 40 healthy adult blood and 20 patient synovial
fluid (SF) samples were included in this study. cDNA was
generated from total RNA extracted from Tempus vacu-
tainers using whole blood or SF. CD4, CD8b and GAPDH
transcripts were measured by qPCR. Simultaneously per-
ipheral blood mononuclear cells (PBMC) and SFMC were
isolated using density gradient centrifugation and stained
with CD3, CD4, CD8b and CD8a for flow cytometry.
Results
Validating previous results, the CD4/CD8 T-cell ratio
measured by flow cytometry was significantly lower in
SFMC than in healthy adult PBMC. Measurement of
CD4/8 ratio by qPCR and flow cytometry showed some
correlation in healthy adult PBMC. However CD4/CD8
ratios by qPCR were higher in SF samples compared to
healthy control PBMC. This was driven by increased CD4
measurements by qPCR in SF samples.
Conclusion
Taken together these data show a trend to a correlation of
qPCR and flow cytometry methods in healthy adult con-
trol samples. The increased level of CD4 transcript in SF
measured by qPCR might well be due to the abundance of
neutrophils and monocytes, which are/may be discarded
during preparation of SFMC for flow cytometry. Whether
qPCR measurement of CD4/CD8 ratio can be used as a
predictive biomarker for severity of disease course in
O-JIA remains to be seen, and will be assessed once clini-
cal follow-up data is available.
Disclosure of interest
None declared.
Authors’ details
1Rheumatology department, UCL Institute of Child Health, London, UK. 2Life
Siences, HAN University of Applied Sciences, Nijmegen, The Netherlands.
3Immunobiology department, UCL Institute of Child Health, London, UK.
1Rheumatology department, UCL Institute of Child Health, London, UK
Full list of author information is available at the end of the article
De Leeuw et al. Pediatric Rheumatology 2013, 11(Suppl 2):P15
http://www.ped-rheum.com/content/11/S2/P15
© 2013 De Leeuw 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.
Published: 5 December 2013
doi:10.1186/1546-0096-11-S2-P15
Cite this article as: De Leeuw et al.: PReS-FINAL-1018: Can the CD4/
CD8b ratio be used as a predictive biomarker in extended-to-be
oligoarticular JIA? Pediatric Rheumatology 2013 11(Suppl 2):P15.
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De Leeuw et al. Pediatric Rheumatology 2013, 11(Suppl 2):P15
http://www.ped-rheum.com/content/11/S2/P15
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PReS-FINAL-1018: Can the CD4/CD8β ratio be used as a predictive biomarker in extended-to-be oligoarticular JIA?

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PReS-FINAL-1018: Can the CD4/CD8β ratio be used as a predictive biomarker in extended-to-be oligoarticular JIA?

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