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Blood, Vol. 105, Issue 12, 4583-4589, June 15, 2005
Therapeutic potential of a tumor-specific, MHC-unrestricted T-cell receptor expressed on effector cells of the innate and the adaptive immune system through bone marrow transduction and immune reconstitution
Blood Alajez et al.
105: 4583
Supplemental materials for: Alajez et al, Vol 105, Issue 12, 4583-4589
Files in this Data Supplement:
- Figure S1. RT-PCR analyses of scTCR expression in transduced BM cells and in splenocytes from reconstituted SCID mice (JPG, 319 KB)
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(A) Expression of scTCR mRNA in transduced BM cells 72 hours after transduction. (B) Expression of scTCR mRNA in splenocytes 60 days after reconstitution. M indicates a 1-kb DNA molecular- weight marker;  -actin, RT-PCR control.
- Figure S2. Long-term expression of scTCR in mice reconstituted with scTCR-transduced BM cells (JPG, 245 KB)
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Control (A) or scTCR-reconstituted (B) mice were bled 7 months after reconstitution and PBMC were stained for GR-1. Cells were gated on GR-1+ and were plotted against EGFP.
- Movie S1. Live imaging microscopy of intracellular Ca2+ changes in control-transfected J.RT3-T3.5 cells (MOV, 633 KB)
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Cells were labeled with fura-2 AM (molecular probes, Eugene, Oregon) and stimulated with HPAF, a human MUC1+ pancreatic tumor. Intracellular Ca2+ levels were monitored. Blue color represents increased intracellular calcium level above background level. Imaging was done every 10 seconds for a total of 10 minutes. Images were taken using a Zeiss live imaging microscope (Thornwood, New York) at the Center for Biologic Imaging, University of Pittsburgh.
- Movie S2. Live imaging microscopy of intracellular Ca2+ changes in J.RT3-T3.5 cells transfected with the TCR α and β chains from the MA CTL clone (MOV, 1.4 MB)
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Cells were labeled, stimulated, and imaged as described for Movie S1.
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