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. 2011 Jun 13;8(5):415–423. doi: 10.1038/cmi.2011.18

Figure 5.

Figure 5

Tolerant 2C cells do not respond to IL-15 or IL-7. (a, b) Comparison of IL-7Rα and IL-15Rβ in 2C T cells from prostate, spleen, PDLN and lung of B6, B6-SIY, TRAMP and TRP-SIY mice. Sixty dpi, cells from the indicated tissues of B6, B6-SIY, TRAMP and TRP-SIY mice were stained for 2C TCR, CD8 and IL-15Rβ or IL-7Rα followed by flow cytometry analysis. Histograms show representative IL-7Rα or IL-15Rβ expression of CD8+ 2C T cells from at least two separate experiments. Naive 2C T cells from spleen were analyzed at the same time for comparison. (c) Tolerant 2C T cells do not respond to IL-15 and IL-7. Cells from TRP-SIY prostates (∼47 dpi) were analyzed for phosphorylated Stat5 (pStat5) as described in the section on ‘Materials and methods'. Histograms show pStat5 level of Thy1.1+CD8+ 2C cells. Naive 2C T-cell response to IL-7 and IL-15 are shown as control. (d) IL-15 and IL-7 transcripts are not detected in prostate tumor tissue. RNA was extracted from prostates of TRP-SIY and TRAMP mice and the transcript levels of IL-7, IL-15 and GAPDH were determined using quantitative real-time PCR. RNA from the bone marrow of B6 mice was used as control. The IL-7 and IL-15 levels were normalized to GAPDH levels of the same tissue. The relative expression levels of IL-7 and IL-15 in different tissues are shown. Error bars represent the standard deviation from duplicate (B6) or triplicate (TRAMP and TRP-SIY) samples. B6, C57BL/6; dpi, days post-infection; PDLN, prostate draining lymph nodes; SIY, SIYRYYGL; TRAMP, TRansgenic Adenocarcinoma of the Mouse Prostate; TRP-SIY, TRAMP mice expressing SIY antigen in prostate tumor tissue.