Figure 6. PTCL in ITK-SYKCD4-Cre mice. (A) Disruption of the splenic architecture with highly proliferative cells in ITK-SYKCD4-Cre mice was revealed by hematoxylin and eosin (H&E) staining and immunohistochemistry with anti–Ki-67 antibodies. Bars: (black) 1 mm; (white) 50 µm. Data shown are representative of five diseased ITK-SYKCD4-Cre mice analyzed. (B) Splenic cells from diseased ITK-SYKCD4-Cre mice were stained against CD4 and CD8 and analyzed by FACS. Selected examples of each type of T cell expansion from a total number of 40 mice analyzed are shown. (C) Bone marrow cell preparations from diseased ITK-SYKCD4-Cre or control (CD4-Cre) mice were stained with antibodies against TCR-β. The frequency of eGFP+ T cells is indicated. Data are representative of five independent experiments with a total number of 15 mice per genotype analyzed. (D) Solid organ infiltration of abnormal CD3+ T cells. Tissue sections from kidney (KID), liver (LIV), and lung (LNG) of affected ITK-SYKCD4-Cre animals were stained with H&E. Immunohistochemistry with anti-CD3 antibodies was additionally performed. Bars: (black) 200 µm; (white), 1 mm. Data shown are representative of five diseased ITK-SYKCD4-Cre mice analyzed. (E) Selective expansion of distinct T cell clones. Single cell suspensions from spleen (SPL), kidney (KID), and liver (LIV) of three individual mice were stained with antibodies against CD4, CD8, and a panel of TCR-Vβ chain–specific antibodies (see Materials and methods). Frequencies of CD4+ or CD8+ cells expressing the indicated TCR-Vβ chains in the spleen of control mice (open histograms) or frequencies of cells expressing the indicated TCR-Vβ chains in spleen, kidney, and liver from diseased ITK-SYKCD4-Cre mice (gray histograms) are shown. (F) Genescan analysis for TCR gene rearrangements. Representative fragment size distributions of fluorochrome-labeled PCR products of the Dβ2/Jβ2 junction of a control and a diseased ITK-SYKCD4-Cre mouse are shown. Data shown are representative of three mice per genotype analyzed. Data in A–F are from ITK-SYKCD4-Cre mice that were older than 12 wk and showed disease symptoms.
Figure 6.

PTCL in ITK-SYKCD4-Cre mice. (A) Disruption of the splenic architecture with highly proliferative cells in ITK-SYKCD4-Cre mice was revealed by hematoxylin and eosin (H&E) staining and immunohistochemistry with anti–Ki-67 antibodies. Bars: (black) 1 mm; (white) 50 µm. Data shown are representative of five diseased ITK-SYKCD4-Cre mice analyzed. (B) Splenic cells from diseased ITK-SYKCD4-Cre mice were stained against CD4 and CD8 and analyzed by FACS. Selected examples of each type of T cell expansion from a total number of 40 mice analyzed are shown. (C) Bone marrow cell preparations from diseased ITK-SYKCD4-Cre or control (CD4-Cre) mice were stained with antibodies against TCR-β. The frequency of eGFP+ T cells is indicated. Data are representative of five independent experiments with a total number of 15 mice per genotype analyzed. (D) Solid organ infiltration of abnormal CD3+ T cells. Tissue sections from kidney (KID), liver (LIV), and lung (LNG) of affected ITK-SYKCD4-Cre animals were stained with H&E. Immunohistochemistry with anti-CD3 antibodies was additionally performed. Bars: (black) 200 µm; (white), 1 mm. Data shown are representative of five diseased ITK-SYKCD4-Cre mice analyzed. (E) Selective expansion of distinct T cell clones. Single cell suspensions from spleen (SPL), kidney (KID), and liver (LIV) of three individual mice were stained with antibodies against CD4, CD8, and a panel of TCR-Vβ chain–specific antibodies (see Materials and methods). Frequencies of CD4+ or CD8+ cells expressing the indicated TCR-Vβ chains in the spleen of control mice (open histograms) or frequencies of cells expressing the indicated TCR-Vβ chains in spleen, kidney, and liver from diseased ITK-SYKCD4-Cre mice (gray histograms) are shown. (F) Genescan analysis for TCR gene rearrangements. Representative fragment size distributions of fluorochrome-labeled PCR products of the Dβ2/Jβ2 junction of a control and a diseased ITK-SYKCD4-Cre mouse are shown. Data shown are representative of three mice per genotype analyzed. Data in A–F are from ITK-SYKCD4-Cre mice that were older than 12 wk and showed disease symptoms.

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