Peripheral CD4CD8 Double Positive T Cells with a Distinct Helper Cytokine Profile Are Increased in Rheumatoid Arthritis

Dagmar Quandt,Roger Scholz,Kathrin Rothe,Ulf Wagner,Christoph W Baerwald,Oliver Frey

doi:10.1371/journal.pone.0093293

Abstract

Peripheral CD4CD8 double positive (DP) T cells have been reported to play a role in several autoimmune diseases, virus infections and cancer. In rheumatoid arthritis (RA), both CD4 and CD8 single positive (SP) T cells are known to be involved in the pathogenesis, but the role of peripheral CD4CD8 DP T cells has not been investigated in detail. Anti cyclic citrullinated antibodies (ACPA) positive and ACPA negative RA patients, patients with systemic lupus erythematodes (SLE) and age matched healthy donors (HD) were enrolled in the analysis. The frequencies and phenotype of DP T cells in PBMC were investigated. In addition, DP T cells were quantified in biopsies from rheumatoid synovium. After in vitro restimulation, the cytokine production of DP T cells was investigated in cultures of PBMC. CMV specific cytokine secretion as well as proliferation was analyzed following antigen specific restimulation after an appropriate culture duration. DP T cells were found more frequently in RA patients than in healthy controls or patients with SLE. These DP T cells express αβ TCRs, are of a memory phenotype and share features of both CD4 as well as CD8 SP T cells. Importantly, DP T cells were found to also be present in the rheumatoid synovium. Further characterization of DP T cells from RA patients revealed increased production of IL-21 and IL-4, implying a possible role as T helper cells. In addition, DP T cells in RA seem to contribute to the inflammatory process, because they produce significantly more IFNγ than counterparts from HD and are increased in CMV+ RA patients. Given their capacity to produce a variety of cytokines (IL4, IL21 and IFNγ), their association with ACPA positive RA and their presence in the synovium, we suggest an important role of double positive T cells in the pathogenesis of rheumatoid arthritis.

Highlights

Peripheral CD4CD8 double positive (DP) T cells have first been identified more than 20 years ago
Total CD4CD8 DP T cells were quantified as percentage of total CD3+ T cells in PBMC, and three subpopulations (CD4hiCD8lo, CD4hiCD8hi and CD4luCD8hi) according to the level of coreceptor expression could be distinguished as shown in figure 1A
CD4hiCD8lo DP T cells are the most prominent of the DP cell populations and were found to be significantly more frequent in PBMC from anti-citrullinated peptide antibodies (ACPA) positive rheumatoid arthritis (RA) patients when compared to healthy controls, systemic lupus erythematodes (SLE) patients or ACPA negative RA patients

Summary

Introduction

Peripheral CD4CD8 double positive (DP) T cells have first been identified more than 20 years ago. Like their progenitors in the thymus, they express the coreceptors CD4 and CD8 simultaneously, but in contrast to immature double positive thymocytes, they show varying degrees of coreceptor expression, and display a memory phenotype but no markers of recent thymic emigrants [2,3,4]. The peripheral CD4+ T cell pool in RA is characterized by several alterations including a paucity of naıve T cells and recent thymic emigrants, an increased memory pool and a global loss of T cell receptor diversity accompanied by large clonal expansions [12,13]

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