W.H. Abdulahad, Y.M. van der Geld, C.A. Stegeman, C.G.M. Kallenberg

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Persistent expansion of CD4+ effector memory T cells in Wegener's granulomatosis W.H. Abdulahad, Y.M. van der Geld, C.A. Stegeman, C.G.M. Kallenberg Kidney International Volume 70, Issue 5, Pages 938-947 (September 2006) DOI: 10.1038/sj.ki.5001670 Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 1 CD4 and CD8 naive and memory T-cell subpopulations in peripheral blood of WG-patients in remission (R-WG, n=40), active WG-patients (A-WG, n=17), and HCs (n=21). (a) The percentages of total memory cells (CD45RO+) within the CD4+ and CD8+ T-cell population in peripheral blood of HC, R-WG, and A-WG patients. (b) Representative example of multicolor flow cytometric analysis of TNaive (CCR7+CD45RO−), TCM (CCR7+CD45RO+), TEM (CCR7−CD45RO+) and TtEM (CCR7−CD45RO−) after gating on CD4 or CD8 T cells from a representative R-WG patient and a sex- and age-matched HC. The fraction of CD4+TEM is increased and the fraction of CD4+TNaive is decreased in this R-WG patient as compared to HC, whereas no differences are seen within CD8 T-cell subpopulations. (c) Percentages of TNaive, TCM, TEM, and TtEM subpopulations within the CD4+ T cells (left) and CD8+ T cells (right). (d) Ratios of the absolute counts, both for CD4+ and CD8+ T-cell subpopulations, of total memory/TNaive, TEM/TCM, and TEM/TNaive. The open symbols within the patients groups represent localized disease, and the solid symbols represent generalized WG. Bars represent mean percentage. The P-values were calculated using the non-parametric Mann–Whitney U-test. Kidney International 2006 70, 938-947DOI: (10.1038/sj.ki.5001670) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 2 Longitudinal analysis of CD4+TEM in WG-patients and HCs. Percentages (a) of CD4+TEM and ratios of absolute counts (b) of CD4+TEM/TNaive were analyzed at two time points, 15±2 months apart, in HCs (n=5) and in R-WG-patients during stable remission (n=5) as well as in patients during active disease and remission (n=6). Horizontal lines represent the median values. The P-values were calculated using the Wilcoxon matched-pairs test. Kidney International 2006 70, 938-947DOI: (10.1038/sj.ki.5001670) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 3 Circulating FoxP3+TEM and FoxP3−TEM among CD4+T-cells in WG-patients and HCs. Percentages of FoxP3-positive and FoxP3-negative CD4+TEM cells (a), and the ratios of FoxP3−CD4+TEM/FoxP3+CD4+TEM (b) in WG-patients in remission (R-WG, n=5), patients with active WG (A-WG, n=5), and age-matched HCs (n=5). FoxP3 expression was determined using an intracellular staining procedure on PBMCs and data were analyzed on FACS-Calibur. Horizontal lines represent median values, and P-values were calculated using the non-parametric Mann–Whitney U-test. Kidney International 2006 70, 938-947DOI: (10.1038/sj.ki.5001670) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 4 Cell surface expression of IL-18Rα and ST2L markers. Expression of IL-18Rα and ST2L on (a) total CD4+ T cells and (b) CD4 central (TCM) and effector (TEM) memory T cells in peripheral blood of WG-patients in remission (R-WG), active WG-patients (A-WG) and HCs. Horizontal lines represent mean percentage. The open symbols within the patients group represent localized disease, and the solid symbols represent generalized WG. The P-values were calculated using the non-parametric Mann–Whitney U-test. Kidney International 2006 70, 938-947DOI: (10.1038/sj.ki.5001670) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 5 Correlation between age (in years) and the percentage of CD4+TNaive, TCM, and TEM cell subpopulations. (a) CD4+TEM cells increase significantly, whereas CD4+TNaive cells show a significant decrease with age in WG-patients in remission. (b) No such correlations are seen in healthy individuals. The r2 and P-values were calculated using linear regression. Kidney International 2006 70, 938-947DOI: (10.1038/sj.ki.5001670) Copyright © 2006 International Society of Nephrology Terms and Conditions