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Alvin W. L. Schadenberg, MD, Theo van den Broek, MD, Marten A

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1 Differential homeostatic dynamics of human regulatory T-cell subsets following neonatal thymectomy 
Alvin W.L. Schadenberg, MD, Theo van den Broek, MD, Marten A. Siemelink, MD, Selma O. Algra, MD, Petrus R. de Jong, MD, Nicolaas J.G. Jansen, MD, PhD, Berent J. Prakken, MD, PhD, Femke van Wijk, PhD  Journal of Allergy and Clinical Immunology  Volume 133, Issue 1, Pages e6 (January 2014) DOI: /j.jaci Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 CD4+ and FOXP3+ T-cell dynamics and expression of CD31 and Ki67 after thymectomy. A, CD4+ T-cell count. B, FOXP3+CD4+ Treg-cell count. C, Percentage of CD31+ (FOXP3+) Treg cells. D, Percentage of FOXP3+ (CD4+) cells. E, Percentage of FOXP3− and FOXP3+ T cells in cell cycle (Ki67+) in thymectomized subjects (TX) (▲) and age-matched controls (HC) (○). Horizontal line represents median value per group. *P < .05 and **P < .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 FOXP3+ subpopulation dynamics after thymectomy. A, Percentage of the 3 FOXP3+ subpopulations within CD4 T-cell population. B, Cell count of the 2 FOXP3+ Treg-cell populations in healthy controls (HC) (○) and thymectomized patients (TX) (▲), and ratio of fraction ii:i. C, Expression of Ki67 per FOXP3+ Treg-cell population. n.s., No significant difference. **P < .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 FOXP3 percentages and counts in the first years after neonatal thymectomy. A, Absolute CD4+ T-cell counts per microliter of blood. B, FOXP3+CD4+ Treg-cell counts per microliter of blood. C, Percentage of FOXP3+ cells in CD4+ T-cell populations. ▲, values after thymectomy; Δ, samples taken just before thymectomy; ○, healthy controls. Lines connect longitudinal samples. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Gating strategy of subpopulations of FOXP3+ T cells. FOXP3+ subpopulations after gating for CD4+ lymphocytes. Fraction I, CD45RO−FOXP3low; fraction II, CD45RO+FOXP3high; fraction III, CD45RO+FOXP3low. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 CD45RO−Foxp3+ T cells represent CD45RA+Foxp3+ T cells. A, Dot plot of CD45RO (memory) and CD45RA (naive) expression on CD3+CD4+ T cells. B, Dot plot of CD45RO and CD45RA expression on CD3+CD4+Foxp3 (Treg) cells. C, Expression of CD45RA+ in the CD45RO−FOXP3+ subpopulation. Healthy control group (HC), n = 7, and thymectomized patients (TX), n = 8. Data represented as mean percentage ± SD. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Subgroup analysis of naive and memory Treg-cell populations in 0- to 6-month-old and more than 6-month-old subjects. CD45RO−FOXP3low naive Treg (A) and CD45RO+FOXP3high memory Treg-cell (B) numbers in the subgroups less than 6 months and more than 6 months of age in healthy controls (HC) and thymectomized patients (TX). Data represented as median, 25% and 75% percentile boxes, and range. *P < .05. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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