Immunological milieu in the peritoneal cavity at laparotomy for gastric cancer

Figure 1 Co-staining with foxp3 and CD25 for CD4⁺ T cells. High correlation was shown between both populations.

Figure 2 Analysis of lymphocyte populations in peripheral blood and the peritoneal cavity. A: The gating and counting of CD4⁺ CD25^high T cell population by flow cytometry; B: The percentage of CD4⁺ CD25^high T cells in the CD4⁺ T cell population in peripheral blood and peritoneal lavage of patients at each stage of gastric cancer and control patients. Data are presented as the mean ± SD.

Figure 3 Cytokine production by lymphocytes. A: The gating and counting of the IFN-γ producing cell population by flow cytometry; B: The percentage of IFN-γ producing cells in the CD3⁺ cell population stimulated with PMA + ionomycin in peripheral blood and peritoneal lavage of patients at each stage of gastric cancer and control patients Data are presented as the mean ± SD. The statistical analysis was performed by the Kruskal-Wallis test. After gating of CD3+ T cells, 10  000 events were analyzed. The production of IFN-γ in the peritoneal cavity was higher than that in the peripheral blood. The ratio of IFN-γ production cells in the peritoneal lavage was significantly lower in patients with advanced-stage than in controls [control vs stage IV: 51.1 (35.1-67.1) vs 10.7 (2.6-22.1), ^aP < 0.05]; C: The gating and counting of the IL-10 producing cell population by flow cytometry; D: The percentage of IL-10 producing cells in the CD3⁺ cells stimulated with PMA + ionomycin in peripheral blood and peritoneal lavage of patients at each stage of gastric cancer and control patients. Data are presented as the mean ± SD. The ratio of IL-10 producing cells in peripheral blood and intra-peritoneal lymphocytes was significantly higher in patients at advanced stage than in controls [control vs stage IV: 6.1 (3.94-8.25) vs 40.7 (18.35-63.0), ^aP < 0.05].

Figure 4 Cytokine assays of intra-peritoneal lymphocytes after co-cultivation with self-CD4⁺CD25 ^highT cells. A: IFN-γ production in intra-peritoneal lymphocytes co-cultivated with self- CD4⁺ CD25^high T cells; B: Either CD4⁺ CD25^high T cells or CD4⁺ CD25^- T cells.