Fig. 9.
Fig. 9. IgH, TCR, and. / neo gene rearrangements in FGFR3-induced tumors and cell lines. (A) Top panel: Southern blot analysis ofEcoRI-digested spleen cell genomic DNA and B220+cell line DNA with an IgH JH probe. As controls, a normal littermate is shown in lane 1 and a recipient of MINV-transduced BM in lane 2 (MINV). WT represents mice receiving MFINV-WT transplants (lane 3) and TD represents mice receiving MFINV-TD transplants. Mice in each experiment were numbered 1 through 5; passage number was also labeled 1 through 5. Thus cells derived from the second and fifth primary MFINV-TD BMT recipients (labeled TD2 and TD5, respectively) were injected into the secondary mice denoted TD2.2, TD2.3, TD2.4, or TD5.1 and TD5.2. Cells from a secondary animal passaged into a fourth generation recipient are labeled TD2.2.1.2. Cell lines derived from animals are designated TD2.1-L and TD2.2.1.2-L. JHrearrangements of the IgH locus were detected only in the fourth generation recipient TD2.2.1.2 as well as the TD 2.2.1.2-L and TD2.1-L cell lines. Middle panel: Southern blot analysis ofPvuII-digested spleen cell genomic DNA from MFINV-TD recipients with a TCR Cβ2 probe. DNA derived from the second and fifth primary FGFR3-TD BM recipients (labeled TD2 and TD5, respectively) were injected into the secondary mice denoted TD2.2, TD2.4, TD5.1, and TD5.2. TCR gene rearrangements were detected in the TD2.2 and TD5.1 samples, both from mice displaying abnormal T-cell populations by flow cytometry. Bottom panel: Genomic DNA digested with EcoRI (which cuts MFINV-TD once) and hybridized with a neo probe was used to enumerate the proviral copy number and determine the clonality of the tumors. MINV represents vector only control mice. TD represents FGFR3-TD BMT mice. Cells from mouse TD2 or TD5 were injected into secondary and tertiary recipients denoted TD2.2, TD2.2.1, or TD5.1 and TD5.2, respectively. In both cases, a predominant clone within a diffuse background was observed in early recipients, whereas multiple bands representing unique clones emerged in later generation mice receiving serial transplants. (B) JH and neo were hybridized as above to MINV control animals and to the primary (WT6, WT1, WT2), secondary (WT1.1, WT2.1), tertiary (WT1.1.1, WT2.1.1), and quarternary mice receiving FGFR3-WT transplants (WT1.1.1.1). No emerging clonal populations were observed in any of the mice.

IgH, TCR, and

neo gene rearrangements in FGFR3-induced tumors and cell lines. (A) Top panel: Southern blot analysis ofEcoRI-digested spleen cell genomic DNA and B220+cell line DNA with an IgH JH probe. As controls, a normal littermate is shown in lane 1 and a recipient of MINV-transduced BM in lane 2 (MINV). WT represents mice receiving MFINV-WT transplants (lane 3) and TD represents mice receiving MFINV-TD transplants. Mice in each experiment were numbered 1 through 5; passage number was also labeled 1 through 5. Thus cells derived from the second and fifth primary MFINV-TD BMT recipients (labeled TD2 and TD5, respectively) were injected into the secondary mice denoted TD2.2, TD2.3, TD2.4, or TD5.1 and TD5.2. Cells from a secondary animal passaged into a fourth generation recipient are labeled TD2.2.1.2. Cell lines derived from animals are designated TD2.1-L and TD2.2.1.2-L. JHrearrangements of the IgH locus were detected only in the fourth generation recipient TD2.2.1.2 as well as the TD 2.2.1.2-L and TD2.1-L cell lines. Middle panel: Southern blot analysis ofPvuII-digested spleen cell genomic DNA from MFINV-TD recipients with a TCR Cβ2 probe. DNA derived from the second and fifth primary FGFR3-TD BM recipients (labeled TD2 and TD5, respectively) were injected into the secondary mice denoted TD2.2, TD2.4, TD5.1, and TD5.2. TCR gene rearrangements were detected in the TD2.2 and TD5.1 samples, both from mice displaying abnormal T-cell populations by flow cytometry. Bottom panel: Genomic DNA digested with EcoRI (which cuts MFINV-TD once) and hybridized with a neo probe was used to enumerate the proviral copy number and determine the clonality of the tumors. MINV represents vector only control mice. TD represents FGFR3-TD BMT mice. Cells from mouse TD2 or TD5 were injected into secondary and tertiary recipients denoted TD2.2, TD2.2.1, or TD5.1 and TD5.2, respectively. In both cases, a predominant clone within a diffuse background was observed in early recipients, whereas multiple bands representing unique clones emerged in later generation mice receiving serial transplants. (B) JH and neo were hybridized as above to MINV control animals and to the primary (WT6, WT1, WT2), secondary (WT1.1, WT2.1), tertiary (WT1.1.1, WT2.1.1), and quarternary mice receiving FGFR3-WT transplants (WT1.1.1.1). No emerging clonal populations were observed in any of the mice.

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