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Blood, Vol. 111, Issue 1, 344-350, January 1, 2008
A Stat5b transgene is capable of inducing CD8+ lymphoblastic lymphoma in the absence of normal TCR/MHC signaling
Blood Bessette et al.
111: 344
Supplemental materials for: Bessette et al
Files in this Data Supplement:
- Figure S1. Stat5b-mediated T-cell expansion in MHC class I deficient (β2m−/−) mice (JPG, 65.6 KB)
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(A) Thymic CD4/CD8 profiles of 8-wk-old  2-m−/− mice with and without the Stat5b transgene and the OT-I TCR transgene. Numbers indicate the percent of cells in each quadrant and total cellularity is indicated above each dot plot. (B) sTCR expression on CD8+ SP cells. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region. (C) Intracellular staining with Anti-Terminal Deoxynucleotidyl Transferase (anti-TdT). This antibody labels normal cortical thymocytes and primitive lymphocytes. Control (polyclonal serum followed by secondary antibody) shown as overlay. Numbers indicate the mean fluorescence intensity (MFI) for the gated CD8+ SP population.
- Figure S2. Stat5b-mediated T-cell expansion on the TCRα−/− background (JPG, 48.1 KB)
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(A) Thymic CD4/CD8 profiles of an 8-wk-old Stat5b TG/ TCR −/− mouse with a TCR+/− littermate serving as a control. Numbers indicate the percent of cells in each quadrant and total cellularity is indicated above each dot plot. (B-C) Surface sTCR (B) and cytoplasmic cTCR (C) expression in CD8+ SP cells. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region. Note the lack of surface expression on Stat5bTG/ TCR−/− cells but presence of cTCR. (D) Intracellular staining with anti-TdT on CD8+ SP thymocytes. Control (polyclonal serum followed by secondary antibody) shown as overlay. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region.
- Figure S3. T cell receptor beta variable (Vβ) staining in Stat5b/ TCRα−/− lymphoma (JPG, 67.3 KB)
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(A) Thymic and splenic CD4/CD8 profiles of a 14-week-old Stat5b TG/ TCR−/− mouse and a TCR+/− (hetero) control. Numbers indicate the percent of cells in each quadrant and total cellularity is indicated above each dot plot. (B-C) Surface sV11 (B) and cytoplasmic (C) cV11 expression in CD8+ SP cells. Numbers indicate percent of cells within the gated region. Note the lack of surface sV11 expression on Stat5b TG/ TCR−/− cells, with significant cV11 staining (similar to TCR staining in Figure S2). Consistent with clonal expansion of V11 cells, cytoplasmic staining for V 2,3,4,5,6,7,8,9,10,12,13,14,17 was negative (data not shown).
- Figure S4. Stat5b-mediated T-cell expansion on the pre-TCRα−/− background (JPG, 48.2 KB)
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(A) Thymic CD4/CD8 profiles of an 8-wk-old Stat5b TG/ pre-TCR−/− mouse with pre-TCR+/− littermates serving as controls. Numbers indicate the percent of cells in each quadrant and total cellularity is indicated above each dot plot. (B-C) Surface sTCR (B) and cytoplasmic cTCR (C) expression in CD8+ SP cells. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region. (D) Intracellular staining with anti-TdT on CD8+ SP cells. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region. Control (polyclonal serum followed by secondary antibody) shown as overlay.
- Figure S5. Lymphoma in Stat5b TG/ pre-TCRα−/− mice (JPG, 93.8 KB)
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(A) Thymic CD4/CD8 profiles of 17-wk-old TCR−/− mice with and without the Stat5b transgene. Also shown is a Stat5b TG on a heterozygous pre-TCR+/− background as a control without lymphoma. Numbers indicate the percent of cells in each quadrant and total cellularity is indicated above each dot plot. (B) HSA surface expression on CD8+ SP thymocytes in pre-TCR−/− mice. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ SP population as well as percent within the gated region. (C) Splenic CD4/CD8 profiles from the same mice. The total cellularity is shown above each dot plot. Numbers indicate the percent of cells within each quadrant. (D) HSA staining on CD8+ splenocytes. Numbers indicate the mean fluorescence intensity (MFI) for the CD8+ population as well as percent within the gated region. (E) CD44/IL-2R profile gated on CD8+ T cells. Numbers indicated percent CD44hiIL-2Rhi memory phenotype CD8+ cells within gated region and of total splenocytes in parentheses.
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