Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement

Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.


Shared' T-Cell Clones Identified in Lymph Nodes and Synovial Tissue of Early Rheumatoid Arthritis Patients Using a Novel High Throughput Sequencing Technology.

Klarenbeek4,  Paul L., de Hair4,  Marjolein J., Alivernini4,  Stefano, van de Sande4,  Marleen G., Doorenspleet4,  Marieke E., van Schaik3,  Barbera D., Berger2,  Ferco H.

Academic Med Ctr/Univ of Amsterdam, Amsterdam, The Netherlands
Dept of Radiology, Academic Medical Center-University of Amsterdam
Dept. of Clin. Epidemiology, Biostatistics and Bioinformatics, Academic Medical Center-University of Amsterdam
Div of Clin Immunology & Rheumatology, Academic Medical Center-University of Amsterdam, Amsterdam, The Netherlands
Lab. For Genome Analysis, Academic Medical Center-University of Amsterdam

Background:

T-cells are likely to play important roles in the pathogenesis of rheumatoid arthritis (RA), but the role of T-cell clones remains to be elucidated. Although the synovial tissue (ST) is the primary target tissue in RA, previous observations from our group suggested that other sites might also be involved in the early phases. As a primary site for adaptive immune responses the lymph node (LN) is a likely candidate. Previously, we demonstrated, using newly developed High Throughput Sequencing (HTS) technology, that T-cell clones are present in the ST of RA-patients, and that the T-cell repertoire in the ST is markedly different from that in the peripheral blood (PB). Here, for the first time, we analyzed paired samples from LN, ST and PB to study T-cell clones in early RA.

Objectives:

Investigate paired LN, ST, and PB samples of early RA patients for presence of T-cell clones to analyze the relation between these compartments with regard to adaptive immune responses.

Methods:

mRNA was isolated from paired LN, ST and PB samples of 2 DMARD-naive anti-CCP+ RA-patients (pt) (fulfilling ACR-criteria at inclusion; disease duration < 1 year) after arthroscopic ST sampling of an arthritic ankle and ultrasound guided needle biopsy sampling of a regional lymph node. A linear amplification with primers for all V(ariable)-genes of the T-cell receptor b-chain was performed. The amplified products contain the Complementarity Determining Region 3 (CDR3), which can be used as 'fingerprint' for each clone. The samples were processed using a Genome Sequencer (454/Roche) analyzing up to 1 million receptors at once. The frequency of each clone was determined by the CDR3 using bioinformatics algorithms. Clones with a frequency of >=1% were arbitrarily considered as highly expanded.

Results:

The LN samples showed only few highly expanded clones when compared to the ST (2 vs. 25 clones (pt1) and 1 vs 24 (pt2) in LN and ST respectively). Few expanded clones were detected in PB as well (0 (pt1) and 2 (pt2)). Evidence of circulation between the compartments was seen in both patients. E.g. of the 24 highly expanded clones in pt2 9 clones were found in the PB and 8 were found in the LN. Of these, 4 were found both in the LN and the PB. Whereas these shared clones were highly expanded in the ST, they were of low frequency in the PB and LN (0.01–0.09% in LN and 0.01–0.52% in PB). Little overlap was seen in the T-cell repertoire between ST and PB/LN. In pt1 only 6% and 8% of the ST clones were shared with the LN and PB respectively. In pt2 this was 9% for both LN and PB.

Conclusions:

This is the first analysis of T-cell clones in LN samples compared to ST and PB, using the HTS technology. In these patients we found T-cell clones in both the regional LN and the ST. Only few highly expanded clones were found in the LN while multiple highly expanded clones were found in the ST, perhaps due to epitope spreading in the ST. Many of the highly expanded ST clones could be found back in LN and/or PB. The frequencies of these clones in LN/PB were very low suggesting local retention and/or proliferation of these clones in the inflamed ST. Additional studies are underway to further characterize the roles of T- and B-cell clones in early RA and to investigate the role of the LN in RA.

To cite this abstract, please use the following information:
Klarenbeek, Paul L., de Hair, Marjolein J., Alivernini, Stefano, van de Sande, Marleen G., Doorenspleet, Marieke E., van Schaik, Barbera D., et al; Shared' T-Cell Clones Identified in Lymph Nodes and Synovial Tissue of Early Rheumatoid Arthritis Patients Using a Novel High Throughput Sequencing Technology. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :1069
DOI: 10.1002/art.28836

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