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Results: 3

1.
Fig. 1.

Fig. 1. From: Unique risk factors for insertional mutagenesis in a mouse model of XSCID gene therapy.

Incidence of lymphoma in mice undergoing XSCID gene therapy. (A) The lymphoma incidence in the first transplant experiment is plotted for the MSCV-γc vector transplant group (n = 15; filled triangle) and the GFP vector control group (n = 17; open triangle). In both of these series, Arf−/−γc−/− bone marrow cells were transduced and transplanted. The third curve was derived by transplanting mice with Arf−/−γc+/+ bone marrow cells that had been transduced with a MFG-γc vector transplant group (n = 18; filled squares). (B) The lymphoma incidence in a second independent transplant experiment is shown. As in A, the γc vector transplant group (n = 18; filled triangle) and the GFP vector control group (n = 17; open triangle) are shown using Arf−/−γc−/− bone marrow cells. The third curve was obtained from mice transplanted with Arf−/−γc+/+ bone marrow cells that had been transduced with a MSCV-γc vector (n = 19, filled square). Time is shown as weeks after transplant.

Yan Shou, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11730-11735.
2.
Fig. 3.

Fig. 3. From: Unique risk factors for insertional mutagenesis in a mouse model of XSCID gene therapy.

Vector integration site analysis in tumor cells. (A) Southern blot analysis of genomic DNA from bone marrow (BM), spleen (S), and thymus (T) of mice with lymphoma in the γc vector-transduced group. DNA was digested with EcoRI, which cuts once in the vector, and probed for a GFP fragment to identify individual integration events. Genomic DNA from a GFP vector-transduced wild-type recipient was used as a probe control. DNA from a normal mouse (WT) without transplantation is shown as a negative control. (B) EcoRI-digested DNA from three lymphomas (numbers above columns) were analyzed by Southern blot. Fragments are labeled as follows: ∗, Tde-1 integration no. 1; ♦, Tde-1 integration no. 2. (C) Schematic illustration of the two Tde-1 integration sites. The arrowheads show the orientation of the integrated vector, and the arrow shows the transcriptional start site.

Yan Shou, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11730-11735.
3.
Fig. 2.

Fig. 2. From: Unique risk factors for insertional mutagenesis in a mouse model of XSCID gene therapy.

Characterization of lymphomas in XSCID gene therapy mice. (A) Histologic appearance of tumors from the γc group. Note the infiltration of lymphoblastic cells in the thymus (i), spleen (ii), lymph node (iii), and bone marrow (iv). (Aiv) Infiltration of the meninges with tumor cells (arrow). (B) FACS profiles of representative lymphoma cells from a GFP control case (Left) and γc vector case (Right). Cells from the control tumor were CD45.1-positive and GFP-negative (Upper Left), demonstrating that they were not transduced and were derived from the transplant recipient. In contrast, lymphoma cells from the γc group were CD45.1-negative and GFP-positive, showing that they were derived from transduced donor cells. In both cases, lymphoma cells were CD4+(Lower, y axis). (C) T cell phenotypes of lymphomas from different mice. Lymphocytes were stained with CD4 and CD8 antibodies as shown (Upper). Compared with thymocytes from a normal mouse (Control), lymphomas seen with the γc vector were either CD8+ (#645), CD4+ (#650), or CD4+CD8+ (#664). Lymphomas are highly GFP-positive, as shown (Lower).

Yan Shou, et al. Proc Natl Acad Sci U S A. 2006 August 1;103(31):11730-11735.

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