Long-Term Immune Reconstitution of Naive and Memory T Cell Pools after Haploidentical Hematopoietic Stem Cell Transplantation  Rita I. Azevedo, Maria.

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Long-Term Immune Reconstitution of Naive and Memory T Cell Pools after Haploidentical Hematopoietic Stem Cell Transplantation  Rita I. Azevedo, Maria V.D. Soares, Adriana S. Albuquerque, Rita Tendeiro, Rui S. Soares, Miguel Martins, Dário Ligeiro, Rui M.M. Victorino, João F. Lacerda, Ana E. Sousa  Biology of Blood and Marrow Transplantation  Volume 19, Issue 5, Pages 703-712 (May 2013) DOI: 10.1016/j.bbmt.2013.01.017 Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Frequency of naive and memory subsets within the CD8+ and CD4+ T cell pools after haploidentical HSCT. The frequency of naive (CD45RA+CD27+), central memory (CM; CD45RA−CD27+), effector memory (EM; CD45RA−CD27−), and CD45RA+ EM (EMRA; CD45RA+CD27−) cell subsets was determined within CD8+ (A) and CD4+ (B) T lymphocytes in HSCT recipients, donors, and age-matched control subjects (∗P < .05 between recipients and donors or control subjects and donors). Flow cytometry images from a representative recipient, the respective donor, and age-matched control subject are shown. Biology of Blood and Marrow Transplantation 2013 19, 703-712DOI: (10.1016/j.bbmt.2013.01.017) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Assessment of CD31 expression, sjTREC content, and telomere length after haploidentical HSCT. (A) CD31 expression was used to identify a population enriched in RTEs within naive (CD45RA+CD27+) CD4+ T lymphocytes. Flow cytometry images from a representative recipient, the respective donor, and age-matched control subject are shown. (B) Levels of sjTRECs within PBMCs. Results are expressed as the copy number of sjTRECs per 106 PBMCs. (C) Telomere length measured within CD8+CD27+ and CD27−, as well as within CD4+CD45RA+ and CD45RA− T lymphocytes. Each symbol represents an individual. Mean values are shown as horizontal lines. ∗P < .05. Biology of Blood and Marrow Transplantation 2013 19, 703-712DOI: (10.1016/j.bbmt.2013.01.017) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 TCR Vβ repertoire of CD8+ and CD4+ T lymphocytes after haploidentical HSCT. Spectratyping analysis of the CDR3 VB regions of the CD8+ (A) and CD4+ (B) T cell pools. Spectratypes for each VB family from a representative recipient and the respective donor and age-matched control subject are shown. Flow cytometry analysis of TCR Vβ family usage within naive CD8+ (C) and CD31+ naive CD4+ T cells (D). Bar graphs show mean with standard error of the mean. ∗P < .05, ∗∗P < .01. Biology of Blood and Marrow Transplantation 2013 19, 703-712DOI: (10.1016/j.bbmt.2013.01.017) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Analysis of the Treg pool after haploidentical HSCT. (A) Frequency of Tregs (CD4+Foxp3+) within the CD4+ T cell population. (B) Proportion of naive cells (CD45RA+CD27+) within the Treg pool. (C) Proportion of CD31+ cells within naive Tregs. (D) Frequency of cycling CD31+ naive Tregs, identified by Ki67 expression. Mean values are shown as horizontal lines. Each symbol represents an individual. (E) Flow cytometry analysis of TCR Vβ family usage within Tregs. Bar graph shows mean with standard error of the mean. ∗P < .05, ∗∗P < .01. Biology of Blood and Marrow Transplantation 2013 19, 703-712DOI: (10.1016/j.bbmt.2013.01.017) Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions