B cell subpopulation profile of ACPA− and ACPA+ RA patients. (A) Representative flow cytometric analysis for the identification of Naïve (IgD+CD27−), Non-switched memory (IgD+CD27+), Switched memory (IgD−CD27+), Double negative memory (IgD−CD27−) and transitional (CD24++CD38++) peripheral blood B cells. (B) Frequency of the indicated peripheral blood B cell subpopulations of ACPA (n = 16) and ACPA+ (n = 17) RA patients. (C) Hierarchical clustering of ACPA− and ACPA+ RA patients based on scaled frequency of 4 major B cell subpopulations. Data are presented as mean ± SEM, symbols represent individual samples. Statistical analysis was performed by using two-tailed Mann-Whitney test, p < 0.05 * were considered significant.