1. Cytomegalovirus-Specific Cytotoxic T Lymphocytes Can Be Efficiently Expanded from Granulocyte Colony-Stimulating Factor–Mobilized Hemopoietic Progenitor Cell Products Ex Vivo and Safely Transferred to Stem Cell Transplantation Recipients to Facilitate Immune Reconstitution 
Leighton E. Clancy, Emily Blyth, Renee M. Simms, Kenneth P. Micklethwaite, Chun-Kei K. Ma, Jane S. Burgess, Vicki Antonenas, Peter J. Shaw, David J. Gottlieb 
Biology of Blood and Marrow Transplantation 
Volume 19, Issue 5, Pages (May 2013)
DOI: /j.bbmt
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

2. Figure 1
Expansion and phenotype of CMV-specific T cell cultures from G-HPC apheresis products. (A) Fold increase in total cell number after 14 days and 21 days in culture for G-HPC–derived products (n = 9). (B) Phenotype of cells expanded from G-HPC products at the completion of culture (n = 11). Dots represent individual cultures, and horizontal lines indicate means. (C) Phenotypic comparison of PBMC composition isolated from NMPB (dark gray; n = 15) and G-HPCs (light gray; n = 16). *P ≤.05. (D) Comparison of expansion kinetics of CMV CTLs expanded from NMPD (dark gray; n = 52) or G-HPC (light gray; n = 11) collections. (E) Comparison of cell phenotype of cultures expanded from NMPD (dark gray; n = 33) or G-HPC products (light gray; n = 11); bars represent means. *P ≤ .05.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

3. Figure 2
Direct comparison of CMV CTLs expanded from G-HPCs and NMPB from the same donor. (A) Fold increase in total cell number after 21 days in culture when derived from NMPB (black) or G-HPC products (gray) for 2 donors. (B) Comparison of phenotype of cultures expanded from NMPB (dark gray) or G-HPC products (light gray) for 2 donors. (C) Representative cytotoxicity assay (donor 1) of CMV T cell cultures from NMPB (left) and HPCs (right) against HLA-matched target cells pulsed with an overlapping peptide mix of the CMV pp65 protein (♦), NLV peptide (○), or no antigen control (▲).
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

4. Figure 3
Cytotoxic capacity and alloreactivity of CMV-specific CTL cultures using the Cr51 assay. Potential alloreactivity was measured by specific lysis of pretransplantation recipient-derived target cells coincubated with CTL lines expanded from NMPB (n = 33) or G-HPC products (n = 9) at a CTL:target cell ratio of 20:1. CMV-specific cytotoxicity was measured by specific lysis (E:T 20:1) of target cells loaded with an overlapping peptide mix covering CMV pp65.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

5. Figure 4
Cytokine production by CD3, CD4, and CD8 T cells in response to restimulation with CMV pp65. (A) Representative dot plots (donor 2, G-HPC–derived culture) gating on CD3+ T cells to measure IFN-γ and IL-2 production in the absence of antigen (top row, negative control) and after restimulation with CMV pp65 (bottom row). (B) Production of IFN-γ and IL-2 by CD3+, CD8+, and CD4+ T cells on restimulation with an overlapping peptide mix covering CMV pp65 for cultures expanded from G-HPCs (n = 4). Each line represents the cytokine response from a different culture and shows the change in the percentage of cytokine-expressing cells in the presence (pp65) and absence (neg) of antigen. (C) Comparison of cytokine responses in individual CMV T cell cultures expanded from G-HPC products (●) and NMPB (▲) producing IFN-γ and IL-2 in response to CMV pp65. Horizontal lines indicate means.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions

6. Figure 5
Measurement of immune function in 6 patients over time. Virus-specific immunity measured by enzyme-linked immunospot assay (reported as spot-forming units/105 cells; pp65, blue; adenovirus, black) and quantitative CMV PCR (copies/mL; red).
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )
Copyright © 2013 American Society for Blood and Marrow Transplantation Terms and Conditions