Memory CCR6+CD4+ T cell subpopulations are increased in ACPA+ patients compared to matched ACPA− patients. a Gating strategy for the identification of peripheral blood Th17, Th17.1, Th22 and CCR4/CXCR3 DP cell subpopulations. CCR6+ cells were gated on CD4+CD45RO+CD25− T cells. b Real-time PCR expression analysis for IL-17, IFNγ, RORC and TBX21 in sorted Th1, Th17, Th17.1 and CCR4/CXCR3 DP CCR6+ Th cells obtained from patients with RA (8–10 patients per population). Prior to RNA isolation cells were stimulated with antiCD3/CD28 and cultured for 3 days. c-d Proportions of the indicated CD4+ T cell subpopulations within the total memory CD4+ T cell population (c) and memory CCR6+CD4+ T cell population (d) of 27 ACPA+ and 27 ACPA− patients with RA. For statistical analysis Wilcoxon matched-pairs signed-ranks test was performed (* = p < 0.05).