Floor Weerkamp, MSc, Edwin F. E. de Haas, BSc, Brigitta A. E

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Age-related changes in the cellular composition of the thymus in children  Floor Weerkamp, MSc, Edwin F.E. de Haas, BSc, Brigitta A.E. Naber, BSc, W. Marieke Comans-Bitter, BSc, Ad J.J.C. Bogers, MD, PhD, Jacques J.M. van Dongen, MD, PhD, Frank J.T. Staal, PhD  Journal of Allergy and Clinical Immunology  Volume 115, Issue 4, Pages 834-840 (April 2005) DOI: 10.1016/j.jaci.2004.10.031 Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 1 Thymic cellularity is age-dependent. A, Total thymocyte numbers per thymic lobe in normal (black dots) and diseased (open symbols) children, plotted against the log age of each child. B, Median total thymocyte numbers of healthy children in 6 age groups. Gray bars show 10th and 90th percentiles. Kruskal-Wallis test: P=.011. Journal of Allergy and Clinical Immunology 2005 115, 834-840DOI: (10.1016/j.jaci.2004.10.031) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 2 Distribution of classic subsets is age-dependent. A, Contour plot of a representative thymus sample (age 6 months). In the left panel, CD4−CD8− (DN) cells, CD4+CD8+ (DP) cells, and CD8+4− (CD8 SP) cells can be distinguished. The CD4+8− population can be further subdivided into CD4+3− (ISP) cells and CD4+3+ (CD4 SP) cells (right panel). B, Median values of percentage DN, ISP, DP, and SP within the total thymocyte population in each age group. Significant differences between groups exist for DN cells (P=.002), CD4 SP (P=.03), and CD8 SP (P=.037). C, Percentages of DN cells within the total thymocyte population in normal (black dots) and diseased (open symbols) children vs the log age. PerCP, Peridinin chlorophyll protein. Journal of Allergy and Clinical Immunology 2005 115, 834-840DOI: (10.1016/j.jaci.2004.10.031) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 3 Distribution of early progenitors is age-dependent. A, Contour plot of a representative thymus sample (age 6 months). CD34 and CD1a staining within the DN gate. B, Median percentages of CD34+CD1a−, CD34+CD1a+, and total CD34− cells within the DN population in each age group. Significant differences between groups were found for CD34+CD1a− (P=.039) and CD34+CD1a+ (P=.001). PE, phycoerythrin. Journal of Allergy and Clinical Immunology 2005 115, 834-840DOI: (10.1016/j.jaci.2004.10.031) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 4 TCRαβ+ cells in the thymus. Percentages of TCRαβ+ cells within the DP population in normal (black dots) and diseased (open symbols) children vs the age of each child. Journal of Allergy and Clinical Immunology 2005 115, 834-840DOI: (10.1016/j.jaci.2004.10.031) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

Fig 5 B and NK (precursor) cells in the thymus. A, Median percentage and 10th and 90th percentiles of B and NK cells within the total thymocyte population. Differences between groups were significant for B cells (P=.005), but not for NK cells. B, Thymic tissue sections stained for CD20 (left panel) or isotype control (right panel). Similar results were obtained with a CD19 antibody (not shown). C, Cortex; H, Hassall corpuscle; M, medulla. C, Contour plots of a representative thymus sample stained with CD10 and CD20 within the CD19+ gate (lower left panel) and with CD34 within both the CD19+ gate (lower right panel) and the CD16/56+ gate (upper right panel). D, Representative contour plot of a normal pediatric bone marrow. Expression of CD10 and CD20 within the CD19+ gate. PE, Phycoerythrin; SSC, side scatter; PerCP, peridinin chlorophyll protein. Journal of Allergy and Clinical Immunology 2005 115, 834-840DOI: (10.1016/j.jaci.2004.10.031) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions