T Cells Contribute to Pathological Responses in the Non-Targeted Rat Heart following Irradiation of the Kidneys

Figures

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  Figure 1. Image-guided localized kidney radiation technique. Computed tomography images of a representative male rat at 5–6 weeks of age with a 1.5-cm-diameter collimator plan of 5 Gy encompassing both kidneys in equally weighted beams (two laterals) are shown in the axial (A), sagittal (B), and coronal (C) planes. The kidneys are located within the central circle of each image. (D) dose volume histogram demonstrating dose to the kidneys.

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  Figure 2. Immune cell infiltration of the cortex and medulla of kidney after local X-irradiation with 10 Gy in wild-type rat. (A) T cells (CD3+), natural killer cells (CD56+) and macrophages (CD68+) appear as brown color. The horizontal scale bar represents 100 microns. Images are representative data from 3–4 animals per group. Quantification of immune cells in cortex and medulla of kidney. (B) Comparison of changes between 20 days and 120 days. Data are mean + SEM. * = p < 0.05 vs. sham-irradiated control. *** = p < 0.001 vs. sham-irradiated control.

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  Figure 3. Immune cells in heart 120 days after local irradiation of kidneys with 10 Gy of X-rays in wild-type rat. T cells (CD3+), natural killer cells (CD56+), and macrophages (CD68+) appear as brown color. The horizontal scale bar represents 100 microns. Images are representative data from 3–4 animals per group. Quantification of immune cells in heart. Data are mean + SEM. ** = p < 0.01 vs. sham-irradiated control.

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  Figure 4. Effective knock down of circulating T lymphocytes in WAGCD247-/- rats. (A) Representative histogram of peripheral blood mononuclear cell showing CD3+ staining cell counts gated on CD45+ total leukocytes demonstrating depletion of CD3+ T lymphocytes in circulation of male WAGCD247-/- rats at 5–6 weeks of age. (B) Summary of the numbers of CD45+ total leukocytes and CD3+ T lymphocytes in circulation of WAGCD247-/- compared to their wild-type WAG littermate controls. CD3+ lymphocytes were present at very low levels in WAGCD247-/- rats. Data are mean + SEM, n = 3–5 per group. ** = p < 0.01 vs. wild-type. *** = p < 0.001 vs. wild-type.

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  Figure 5. Blood urea nitrogen after local irradiation of the kidneys with 10 Gy of X-rays in wild-type and WAGCD247-/- rats. BUN levels 40–120 days after the start of the study. Data are mean + SEM. n = 6/group. * = p < 0.05 vs. wild-type. ** = p < 0.01 vs. wild-type. *** = p < 0.001 vs. wild-type.

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  Figure 6. Collagen deposition in heart after local irradiation of kidneys with 10 Gy of X-rays in wild-type and WAGCD247-/- rats. (A) Trichrome staining of heart. The horizontal scale bar represents 100 microns. Images are representative data from 3–4 animals per group. (B). Quantification of perivascular cardiac collagen content in hearts of wild type and WAGCD247-/- rats 120 days after the start of the study. Data are mean + SD, n = 6/group.

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  Figure 7. Immune cells in cortex and medulla of kidney 40 days after local irradiation with 10 Gy of X-rays in A. wild-type WAG and B. WAGCD247-/- rats. T cells (CD3+), natural killer cells (CD56+), macrophages (CD68+) and B cells (CD20+) appear as brown color. The horizontal scale bar represents 100 microns. Images are representative data from 3–4 animals per group. Quantification of immune cells in kidney. Data are mean + SEM.

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  Figure 8. Collagen deposition in kidney after local irradiation with 10 Gy of X-rays in wild type WAG and WAGCD247-/- rats. Trichrome staining of kidney. The horizontal scale bar represents 100 microns. Images are representative data from 3–4 animals per group. Quantification of collagen deposition in kidney. Data are mean + SEM.