Reference values for peripheral blood lymphocyte subsets of healthy children in China Yuan Ding, MD, Lina Zhou, MS, Yu Xia, MD, Wei Wang, MS, Ying Wang,

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Reference values for peripheral blood lymphocyte subsets of healthy children in China Yuan Ding, MD, Lina Zhou, MS, Yu Xia, MD, Wei Wang, MS, Ying Wang, MS, Li Li, MS, Zhongxiang Qi, MS, Linqing Zhong, MS, Jinqiao Sun, MD, Wenjing Tang, MD, Fangfang Liang, MS, Haijuan Xiao, MD, Tao Qin, MD, Ying Luo, MD, Xuezhen Zhao, MS, Zhou Shu, MD, Ying Ru, MS, Rongxin Dai, MD, Hong Wang, MS, Yanping Wang, MS, Yongjie Zhang, MS, Suqian Zhang, MS, Cong Gao, MS, Hongqiang Du, MD, Xuan Zhang, MS, Zhaolong Chen, MS, Xiaochuan Wang, MD, PhD, Hongmei Song, MD, Jun Yang, MD, Xiaodong Zhao, MD, PhD Journal of Allergy and Clinical Immunology Volume 142, Issue 3, Pages 970-973.e8 (September 2018) DOI: 10.1016/j.jaci.2018.04.022 Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Lymphocyte subset distribution throughout childhood. A, The relative contribution of CD4 and CD8 T cells, B cells, and NK cells among total lymphocytes during childhood. B, The relative contribution of CD4 T-cell subsets among total CD4 T cells during childhood. C, The relative contribution of CD8 T-cell subsets among total CD8 T cells during childhood. D, The relative contribution of B-cell subsets among total B cells during childhood. TEMRA, Terminally differentiated effector memory CD45RA+ T cells. Journal of Allergy and Clinical Immunology 2018 142, 970-973.e8DOI: (10.1016/j.jaci.2018.04.022) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Age-related changes in the total number of lymphocyte subsets. A, Absolute numbers of lymphocytes, total T cells, B cells, NK cells, and the CD4 T-cell, CD8 T-cell, γδT-cell, and DNT-cell subsets. B, Absolute numbers of CD4 and CD8 T-cell subsets. C, Absolute numbers of B-cell subsets. The curve indicates the lower and upper limits at a confidence level of 0.95. DNT, TCRαβ+ CD4 and CD8 double-negative T cell; TEMRA, terminally differentiated effector memory CD45RA+ T cell. Journal of Allergy and Clinical Immunology 2018 142, 970-973.e8DOI: (10.1016/j.jaci.2018.04.022) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Flow cytometry gating strategy for T-cell and B-cell subsets. A, Stepwise gating strategy for identification of γδT cell (CD3+TCRγδ), naive cytotoxic T cell (CD8 Naive; CD3+CD8+CD45RA+CD27+), terminally differentiated cytotoxic T cell reexpressing CD45RA (CD8 TEMRA; CD3+CD8+CD45RA+CD27−), central memory cytotoxic T cell (CD8 CM; CD8+CD45RA−CD27+), effector memory cytotoxic T cell (CD8 EM; CD3+CD8+CD45RA−CD27−), double-negative T cell (DNT; CD3+TCRαβ+CD4−CD8−), naive helper T cell (CD4 Naive; CD3+CD4+CD45RA+CD27+), terminally differentiated helper T cell reexpressing CD45RA (CD4 TEMRA; CD3+CD4+CD45RA+CD27−), central memory helper T cell (CD4 CM; CD3+CD4+CD45RA−CD27+), and effector memory helper T cell (CD4 EM; CD3+CD4+CD45RA−CD27−). B, Stepwise gating strategy for identification of memory B-cell (CD19+CD27+), naive B-cell (CD19+CD27−IgD+), and transitional B-cell (CD19+CD24++CD38++) subsets and plasmablasts (CD19+CD24−CD38++). SSC, Side scatter. Journal of Allergy and Clinical Immunology 2018 142, 970-973.e8DOI: (10.1016/j.jaci.2018.04.022) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Reference values of absolute counts of lymphocyte subsets in children: Comparison among different countries. A, Comparison of absolute numbers of total T cells during childhood among China (this study), the United States (reference 1), Italy (reference 4), and Germany (reference E9). B, Comparison of absolute numbers of CD4+ T cells during childhood among China, the United States, Italy, and Netherlands (reference 2). C, Comparison of absolute numbers of CD8+T cells during childhood among China, the United States, Italy, and Netherlands. D, Comparison of absolute numbers of B cells during childhood among China, the United States, Italy, and The Netherlands. E, Comparison of absolute numbers of total NK cells during childhood among China, the United States, and Italy. *Data represent as median and 10th to 90th percentiles. Journal of Allergy and Clinical Immunology 2018 142, 970-973.e8DOI: (10.1016/j.jaci.2018.04.022) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions