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Blood, Vol. 110, Issue 7, 2620-2630, October 1, 2007
Epstein Barr virus–specific cytotoxic T lymphocytes expressing the anti-CD30 artificial chimeric T-cell receptor for immunotherapy of Hodgkin disease
Blood Savoldo et al.
110: 2620
Supplemental materials for: Savoldo et al, Vol 110, Issue 7, 2620-2630
Files in this Data Supplement:
- Figure S1. Cytotoxic activity of CD30CAR+ CTLs (JPG, 51.5 KB)
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(A,B) Cytotoxic activity of control and CD30CAR+ Epstein Barr virus-specific cytotixic T-lymphocytes (EBV-CTLs), respectively, from a representative healthy donor. (C,D) Cytotoxic activity of control and CD30CAR+ EBV-CTLs, respectively, generated from a representative HD patient. The graphs illustrate the percent-specific lysis of the CTLs against: autologous LCL lines (♦), HLA-mismatched LCLs (■), the two CD30+ cell lines HDLM-2 (△) and Karpas-299 (*), a CD30-negative target cell line (○), and autologous PHA blasts (●). Shown are data with more detailed titration (from 40:1 to 5:1 Effector:Target ratios).
- Figure S2. Cytotoxic activity of CD30CAR+ CTLs against PHA blasts (JPG, 68.6 KB)
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(A) CD30 expression, evaluated by FACS, on PHA blasts on day 4 and day 8 of culture and of genetically modified blasts stably expressing CD30 (as described in “Patients, materials, and methods, Evaluation of cytotoxic activity”) for one representative of 3 donors. These cells were then labeled with 51Cr and used as targets in a standard cytotoxicity assay. (B) Percent-specific lysis of CD30CAR+ CTLs against these targets. Killing of cells that highly expressed CD30 occurs, while CD30 negative/low cells are not lysed.
- Figure S3. Killing of CD30 overexpressing T cells by CD30CAR (JPG, 61 KB)
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(A) Expression of the CD30 molecule on PHA blasts, T blasts transduced with a retroviral vector to stably express the CD30 antigen and HDLM-2, an HD cell line constitutively expressing the CD30 antigen. (B) Percent of specific killing by control and CD30CAR+ CTLs against these targets as assessed in a standard 51Cr release assay. Shown is the 20:1 Effector:Target ratio. CD30 overexpressing blasts are lysed at a comparable level as HDLM-2.
- Figure S4. Evaluation of bioluminescent signal and expansion of T cells (JPG, 51.8 KB)
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Mice with differing bioluminescence signals at the tumor site were killed, and the cells isolated and stained with anti-human CD3 and CD45 MAbs. (A) Mice with higher bioluminescence intensity (3 × 107 p/s/cm2/sr) also had higher numbers of CD3+ T cells (∼40%). (B) Intermediate intensity (5 × 106 p/s/cm2/sr). The number of T cells in this case was lower (∼20% of CD3+ T cells). (C) A mouse with no measurable bioluminescence (1 × 105) and correspondingly negligible CD3+ T cells (<0.5%).
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