American Society of Hematology

Figure 7

LTα-LTβR signaling enhances expression of CCL19 and CCL21 and supports Eμ-Myc lymphoma progression. (A) A total of 2 × 104 Wt Eμ-Myc GFP tumor cells were intravenously transferred into LTα−/− recipients (n = 4) and Wt controls (n = 5). At day 15, splenic tumor load was compared and expressed as splenic weight. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (B) A total of 2 × 104 Wt (n = 6) or LTα−/− Eμ-Myc tumor cells (n = 7) were intravenously transferred into congenic Wt recipients. At days 15-18 after tumor challenge, tumor load in the spleen was assessed by flow cytometry. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (C) A total of 2 × 104 Wt (n = 7) or LTα−/− Eμ-Myc tumor cells (n = 6) were intravenously transferred into LTa−/− recipients. At days 14-18 after tumor challenge, tumor load in the spleen was expressed as splenic weight. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (D) A total of 5 × 104 Wt (n = 3) or LTα−/− Eμ-Myc tumor cells (n = 3) were intravenously transferred into Rag−/− recipients. At day 17 after tumor challenge, tumor load in the spleen was expressed as splenic weight. Error bars represent mean ± SEM. *P ≤ .05. In one of the 2 experiments performed in panels B to D, splenic CCL19 and CCL21 mRNA expression of tumor challenged Wt or KO recipients was analyzed by quantitative RT-PCR (E-G). Open bars represent basal splenic CCL19 and CCL21 level of Wt (E), LTα−/− (F), and Rag−/− (G) mice. The relative CCL19 and CCL21 RNA levels of tumor-challenged mice (n = 2-4 per group) were calculated as described in “RNA extraction and RT-PCR.” (H) In vivo blockage of the LTβR signaling pathway by treatment of tumor cell recipient mice with 100 μg LTβR-Ig (n = 13) intraperitoneally in 5-day intervals starting on day −1, or control mouse IgG1 (MOCP21; n = 9), as described. Ten to 13 days after tumor challenge (1 × 105 Eμ-Myc GFP tumor cells), tumor load in spleen and LNs was assessed by flow cytometry. Error bars represent mean ± SEM. *P ≤ .05.

LTα-LTβR signaling enhances expression of CCL19 and CCL21 and supports Eμ-Myc lymphoma progression. (A) A total of 2 × 10⁴ Wt Eμ-Myc GFP tumor cells were intravenously transferred into LTα^−/− recipients (n = 4) and Wt controls (n = 5). At day 15, splenic tumor load was compared and expressed as splenic weight. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (B) A total of 2 × 10⁴ Wt (n = 6) or LTα^−/− Eμ-Myc tumor cells (n = 7) were intravenously transferred into congenic Wt recipients. At days 15-18 after tumor challenge, tumor load in the spleen was assessed by flow cytometry. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (C) A total of 2 × 10⁴ Wt (n = 7) or LTα^−/− Eμ-Myc tumor cells (n = 6) were intravenously transferred into LTa^−/− recipients. At days 14-18 after tumor challenge, tumor load in the spleen was expressed as splenic weight. Error bars represent mean ± SD of 2 independent experiments. *P ≤ .05. (D) A total of 5 × 10⁴ Wt (n = 3) or LTα^−/− Eμ-Myc tumor cells (n = 3) were intravenously transferred into Rag^−/− recipients. At day 17 after tumor challenge, tumor load in the spleen was expressed as splenic weight. Error bars represent mean ± SEM. *P ≤ .05. In one of the 2 experiments performed in panels B to D, splenic CCL19 and CCL21 mRNA expression of tumor challenged Wt or KO recipients was analyzed by quantitative RT-PCR (E-G). Open bars represent basal splenic CCL19 and CCL21 level of Wt (E), LTα^−/− (F), and Rag^−/− (G) mice. The relative CCL19 and CCL21 RNA levels of tumor-challenged mice (n = 2-4 per group) were calculated as described in “RNA extraction and RT-PCR.” (H) In vivo blockage of the LTβR signaling pathway by treatment of tumor cell recipient mice with 100 μg LTβR-Ig (n = 13) intraperitoneally in 5-day intervals starting on day −1, or control mouse IgG1 (MOCP21; n = 9), as described. Ten to 13 days after tumor challenge (1 × 10⁵ Eμ-Myc GFP tumor cells), tumor load in spleen and LNs was assessed by flow cytometry. Error bars represent mean ± SEM. *P ≤ .05.

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