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Blood, Vol. 112, Issue 6, 2278-2286, September 15, 2008
Resistance of mature T cells to oncogene transformation
Blood Newrzela et al.
112: 2278
Supplemental materials for: Newrzela et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 61.3 KB)
- Table S1. Phenotype tables of HSC derived tumors (PDF, 81.4 KB)
- Table S2. PANTHERDB and IDDB integration site classification of HSC/HPC derived tumors (PDF, 69.7 KB)
- Table S3. RTCGD hits in HSC/HPC derived tumors (PDF, 42.8 KB)
- Table S4. Genes surrounding integrations in HSC/HPC-derived tumors (PDF, 4.27 MB) -
(A) After LMO2 transduction, (B) after TCL1 transduction, and (C) after dTrkA transduction.
- Table S5. PANTHERDB Biological process site classification of long-term in vivo observed T cells after shotgun-cloning (PDF, 17.8 KB)
- Table S6. Genes surrounding integrations in long-term observed T cells (PDF, 16.8 MB) -
(A) EGFP-transduced, (B) dTrkA high-transduced.
- Figure S1. Histopathology of HSC/HPC derived leukemias/lymphomas (JPG, 161 KB)
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(A) Autopsy of animal LMO2_A2 showing a massively enlarged thymus, occupying the complete upper anterior mediastinum. (B) Thymus of LMO2_A2 with infiltrates of medium-sized lymphoblasts. Macrophages are intermingled, showing a starry sky pattern. (C) (LMO2_A6_Liver) Lymphoblastic infiltrates in portal tracts and adjacent liver parenchyma. (D) Autopsy of animal TCL1_A1 showing enlargement of lymphoreticular organs (lymph nodes, liver and spleen). (E) TCL1_A1 showing medium-sized lymphoblasts undermining bronchial epithelium (arrow). (F) (TCL1_A7_Liver) Lymphoblastic infiltrate in a central vein spreading to liver parenchyma. (G) Massively enlarged spleen of animal ΔTrkAhigh_A5 compared to a spleen of a healthy control animal (below). (H) Spleen of ΔTrkAhigh_A5 with extramedullary hematopoiesis. (I) (ΔTrkAhigh_A10_Brain) Subarachnoidal lymphoblastic infiltrates.
- Figure S2. Survival of animals transplanted with MP91-LMO2 transduced RAG-1−/− deficient HSCs/HPCs (JPG, 28.5 KB)
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Four animals were transplanted with MP91-LMO2 transduced (70%) RAG-1 deficient HSCs/HPCs. Comparable onset of malignancy was observed as with WT cells (Fig. 5A). This indicates that TCR recombination has no major role in the tumorigenesis of LMO2.
- Figure S3. LM-PCR analysis of HSC/HPC derived tumors (JPG, 92.8 KB)
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Ligation-mediated PCR analysis of HSC/HPC derived tumors after transduction with MP91-LMO2 (A), MP91-TCL1 (B) and SF91-ΔTrkAlow (C). Animals ΔTrkA_A3+ and ΔTrkA_A4+ (Figure S3C, lanes 2+3) were from the ΔTrkAhigh group and were used as controls for a highly polyclonal pattern.
- Figure S4. CD4/CD8 ratio before and after transplantation of transduced mature T cells (JPG, 31 KB)
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Mature T cells were transduced with MP91-eGFP. 7 days after isolation, stimulation and transduction most of the transplanted T cells (‘EGFP Input’) showed a CD8+ phenotype. Cells were transplanted into RAG-1−/− deficient mice and after 481 days (‘d481’) of long-term in vivo follw-up the CD4/CD8 ratio had converted to normal. Normal proportions of CD4+ T cells were detected in the lymph nodes, spleen and peripheral blood of recipient mice. n = Number of analyzed animals.
- Figure S5. LM-PCR analysis of mature T cells after long-term follow-up (JPG, 33.3 KB)
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Ligation-mediated PCR analysis of mature T cells on day 481 after transplantation for MP91-eGFP (A), day 518 after transplantation for MP91-LMO2 (B), day 349 after transplantation for MP91-TCL1 (C) and day 292 after transplantation for SF91-ΔTrkAhigh (D). An oligo- to polyclonal integration pattern was preserved during extensive in vivo persistence of mature T cells. ‘I’ indicates the transplanted input and is shown as control for a highly polyclonal pattern.
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