Thymic Activity and T Cell Repertoire Recovery after Autologous Hematopoietic Stem Cell Transplantation Preceded by Myeloablative Radiotherapy or Chemotherapy

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Presentation transcript:

Thymic Activity and T Cell Repertoire Recovery after Autologous Hematopoietic Stem Cell Transplantation Preceded by Myeloablative Radiotherapy or Chemotherapy Magdalena Głowala-Kosińska, Agata Chwieduk, Andrzej Smagur, Wojciech Fidyk, Jacek Najda, Iwona Mitrus, Sebastian Giebel Biology of Blood and Marrow Transplantation Volume 22, Issue 5, Pages 834-842 (May 2016) DOI: 10.1016/j.bbmt.2016.01.014 Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10 Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Cytometric analysis and gating scheme for determination of RTE cells within CD4+ lymphocytes. Four-color cytometry was used to identify RTE cells (CD31+ CD62L+ CD45RA+ CD4+) in peripheral blood samples. (A) Naive T cells were gated within CD4+ cells. (B) CD45RA+ cells coexpressing CD62L and CD31 were gated as RTE cells. This Figure is available in color online at www.bbmt.org. Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 The classification of CDR3 spectra-type profiles of the TCRVβ chain on the basis of relative fluorescent intensity versus the CDR3 length size. (A) Polyclonal repertoire with Gaussian distribution of CDR3 lengths. (B) Polyclonal repertoire with non-Gaussian (skewed) distribution of CDR3 lengths. (C) Oligoclonal repertoire with few dominating CDR3 lengths. (D) Monoclonal repertoire with expression of a single CDR3 length version. Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Loss and recovery of RTE cells and sjTREC copies after HSCT. RTE median values on day 0 and its restoration in consecutive time points after autoHSCT are shown in (A) BEAM and (B) Cy/TBI preconditioned groups. The loss of RTE frequencies on day +100 was more pronounced in the Cy/TBI than BEAM group. On day +365 (in relation to day 0) RTE counts almost normalized in the BEAM group (P = .0758), whereas in the Cy/TBI group RTE counts were still far from baseline values (P = .00086). Similarly, the changes of sjTREC quantities over follow-up are shown in (C) BEAM and (D) for Cy/TBI groups (sjTREC values in log scale). Statistics were analyzed by Wilcoxon test. Statistical significance is marked by asterisks (*P < .05, **P < .0005). Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Representative CDR3 spectra-type profiles of TCR repertoires in 22 Vβ families from 1 patient on day +365 after autoHSCT. The x-axis indicates CDR3 length (every 3 nucleotides), and the y-axis displays arbitrary fluorescence intensity of nested PCR fluorescent products. Most TCRVβ families have altered CDR3 TCR, in most families polyclonal skewed (Vβ2, Vβ4, Vβ5, Vβ7, Vβ12, Vβ14, Vβ15, Vβ17, Vβ18, Vβ20, Vβ23) or restricted (monoclonal, Vβ8, Vβ11, Vβ21, or oligoclonal, Vβ3, Vβ9, Vβ16, Vβ22, Vβ24). Polyclonal Gaussian distribution is represented by Vβ1 and Vβ6 families. Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 T cell repertoire on day +365 after autoHSCT. Three categories of TCR repertoire status were compared between BEAM and Cy/TBI groups: normal spectra-types, skewed spectra-types, and oligoclonal+monoclonal spectra-types. The results are shown as percentages of all 22 spectra-types for each patients. Statistical significance between the 2 groups of patients was calculated by Mann-Whitney U test (*P < .05). Biology of Blood and Marrow Transplantation 2016 22, 834-842DOI: (10.1016/j.bbmt.2016.01.014) Copyright © 2016 American Society for Blood and Marrow Transplantation Terms and Conditions