Figure 8

Tg6F changed the composition of immune cells similarly in the small intestine and lungs providing a working hypothesis/model for the anti-tumorigenic activity of Tg6F in mice. Immune cells were isolated from the lamina propria of the jejunum or from the lungs as described in Materials and Methods and were sorted by flow cytometry for patrolling monocytes or myeloid derived suppressor cells as described in Materials and Methods. (a) Quantification of patrolling monocytes in the lung and in the lamina propria of the jejunum. The results shown are Mean ± SEM. (b) Quantification of myeloid derived suppressor cells in the lung and in the lamina propria of the jejunum. The data shown are Mean ± SEM. (c) Working hypothesis/model for the anti-tumorigenic activity of Tg6F in mice. Tg6F decreases the expression of cholesterol 25-hydroxylase (CH25H) which leads to a decrease in levels of 25-hydroxycholesterol (25-OHC), which in turns leads to decreased levels of osteopontin (Spp1), which leads to decreased levels of myeloid derived suppressor cells (MDSC) in the small intestine. Tg6F also decreases the levels of oxidized phospholipids recognized by the E06 antibody, which leads to increased activity of the Notch pathway with increased levels of Notch1 and Dll4 and Notch2 and Dll1 that increase levels of patrolling monocytes in the small intestine. Additionally, Tg6F alters the expression of cytokines and factors involved in modulating inflammation; TNFα, iNOS and TGFβ expression is decreased, while IL-4, IL-10, IL-12, TLR2, IFNβ, MX-1 and NFKBID expression is increased in the small intestine. The results of these changes in the small intestine are transmitted to the lungs where similar changes occur and result in decreased tumor burden in the lungs following injection of malignant cells into the tail vein of mice with normal immune systems.