Double Umbilical Cord Blood Transplantation: A Study of Early Engraftment Kinetics in Leukocyte Subsets using HLA-Specific Monoclonal Antibodies Judith.

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Double Umbilical Cord Blood Transplantation: A Study of Early Engraftment Kinetics in Leukocyte Subsets using HLA-Specific Monoclonal Antibodies Judith A.E. Somers, Anneke Brand, Yvette van Hensbergen, Arend Mulder, Machteld Oudshoorn, Kees Sintnicolaas, Jan-Willem Gratama, J.H. Frederik Falkenburg, Eric Braakman, Jan J. Cornelissen Biology of Blood and Marrow Transplantation Volume 19, Issue 2, Pages 266-273 (February 2013) DOI: 10.1016/j.bbmt.2012.09.022 Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Patient 9: Flow cytometry of (A) CD45+ cells and (B) CD4+ T cells at day + 11 and day +18 posttransplantation. Both CBUs and recipient are HLA-A2 positive. The predominant CBU is HLA-B60 positive, whereas the nonsustaining CBU is HLA-B35 positive (see Table 1). Samples were stained using anti-HLA-A2 (PE Dy 647) in combination with anti-HLA-B60 (Alexa-488) or anti-HLA-B35 (Alexa-488). The number of recipient cells was calculated as follows: number of recipient cells = total cell number − (HLA-B60 − positive cells + HLA-B35 − positive cells). Biology of Blood and Marrow Transplantation 2013 19, 266-273DOI: (10.1016/j.bbmt.2012.09.022) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Pt. no. 9: Early chimerism in leukocyte subpopulations. Contribution of CBU- and recipient derived cells to peripheral blood cells at day +11, +18, +25 and +32 posttransplantation. Analysis of CD8+ T cells was not possible due to the low number of events. CD4 T cells: CD45+/3+/4+/low SS T lymphocytes; NK cells: CD45+/3-/16/56+/low SS cells; monocytes: CD45+/14+ cells; granulocytes: CD45+/33+ myeloid cells. predominant CBU; non-engrafting CBU; recipient; absolute cell count. Biology of Blood and Marrow Transplantation 2013 19, 266-273DOI: (10.1016/j.bbmt.2012.09.022) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Median absolute cell numbers of CBUs and recipient. (A) Predominant CBU, (B) nonengrafting CBU, and (C) recipient. CD4: CD45+/3+/4+ T lymphocytes (reference values, .49-1.54 × 109/L); CD8: CD45+/3+/8+ T lymphocytes (reference values, .49-1.54 109/L); CD19: CD45+/3-/19+ B cells (reference values, .130-.63 × 109/L); CD56: CD45+/3-/16/56+ NK cells (reference values, .1-.67 × 109/L); CD14: CD45+/14+ monocytes; CD33: CD45+/33+ myeloid cells. Biology of Blood and Marrow Transplantation 2013 19, 266-273DOI: (10.1016/j.bbmt.2012.09.022) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions