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Abstract Number: 1737

Altered Phenotype and Function of Senescent Regulatory T Cells in Rheumatoid Arthritis

Johannes Fessler1, Chrsitine Schwarz1, Anja C. Ficjan1, Rusmir Husic2, Evelyne Höller3, Angelika Lackner1, Winfried B. Graninger4 and Christian Dejaco1,5, 1Rheumatology and Immunology, Medical University Graz, Graz, Austria, 2Rheumatology, Medical University Graz, Graz, Austria, 3Endocrinology, Medical University Graz, Graz, Austria, 4Internal medicine/Rheumatology and Immunology, Medical University Graz, Graz, Austria, 5Department of Rheumatology and Immunology, Medical University Graz, Graz A-8036, Austria

Meeting: 2014 ACR/ARHP Annual Meeting

Keywords: Aging, immunology and rheumatoid arthritis (RA), Senescent Cells, T-Regulatory Cells

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Session Information

Title: T cell Biology in Rheumatoid Arthritis and Other Arthritis

Session Type: Abstract Submissions (ACR)

Background/Purpose

Immunosenescence accompanied by accumulation of senescent T cells  is a hallmark feature in the pathogenesis of rheumatoid arthritis (RA). Here we characterize a novel senescent regulatory T cell (Tregs, CD4+CD28–FoxP3+) subset in RA patients.

Methods

Prospective, cross-sectional study on 35 patients with RA [mean age 58 (±SD 9.5), 71.4% female, SDAI 8.15 (±1.2)] and 25 healthy controls [HC, mean age 56.4 (±6.7), 60% female]. We used flow cytometry to determine the prevalence of senescent CD4+CD28–FoxP3+ T cells and to characterize their phenotype, proliferation, cytokine production and apoptosis. T cell receptor diversity was determined by RT-PCR. In vitro generation of senescent Tregs was performed in cell culture experiments using magnetic bead isolated CD4+CD25+CD127lowTregs and stimulation with anti-CD3/CD28 beads, interleukin (IL) -2 with or without TNF-α (100ng/ml) for 14 days.

Results

Two percent [±2.8] of CD4+ T cells were CD28–FoxP3+ in RA patients whereas this subset was almost absent in HC [0.6 (±0.8), p=0.077]. The number of CD4+CD28+FoxP3+ Tregs was comparable in both groups [28.6 (±18.5) vs. 32.7 (±18), p=0.480]. In vitro assays showed that exposure of CD4+CD28+ Tregs to TNF-α led to a downregulation of CD28 and thus to the CD28–FoxP3+ phenotype.

Surface receptor expression analysis of CD28–FoxP3+ and CD28+FoxP3+ Tregs demonstrated that CD28–FoxP3+ cells expressed higher levels of the regulatory protein PD-1 [17.45% (0-36.4) vs. 5.45% (1.8-13.5), p=0.034], whereas CTLA-4 expression was similar in both subsets. Production of various cytokines including IL-2, IL-4, IL-10, IL-17, TNF-α and IFN-γ was increased in CD28–FoxP3+ compared to CD28+FoxP3+ Tregs [all p<0.05] whereas proliferation rate was lower than in the CD28+ counterparts [50% (0-93.6) non proliferating cells vs. 4.6% (0-30.6), p=0.001]. In contrast, apoptosis induction was higher in CD28–FoxP3+ than in CD28–FoxP3+ Tregs [22.1% (0-30.8) vs. 4.4% (0-7.8), p<0.001]. TCR diversity was also reduced in CD28–FoxP3+ Tregs compared to their CD28+ counterparts [median TCR diversity score: 84 (36-104) vs. 115 (109-125), p=0.037].

Conclusion

We discovered a novel T cell subset which combines both senescent as well as regulatory properties. This subset favors the pro-inflammatory milieu and shows altered phenotype and function compared to normal (non-senescent) Tregs.


Disclosure:

J. Fessler,
None;

C. Schwarz,
None;

A. C. Ficjan,
None;

R. Husic,
None;

E. Höller,
None;

A. Lackner,
None;

W. B. Graninger,
None;

C. Dejaco,

Pfizer, MSD,

2,

Pfizer, MSD, Roche, UCB, BMS, AbbVie,

8.

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