Fig 2: DHX15 is required for control of intestinal inflammation induced by enteric reovirus infection in adult mice in vivo(A) Survival of 5-week-old wild-type Dhx15fl/fl and Dhx15IEC-KO adult mice (n = 10 per strain) after intragastric injection of reovirus T3D strain (1 × 108 plaque-forming units [PFUs] per mouse).(B–D) The wild-type Dhx15fl/fl and Dhx15IEC-KO mice (n = 5 per strain) were inoculated intragastrically with 1 × 108 PFUs reovirus T3D strain. At day 1 post-inoculation, mice were euthanized, and intestine tissues were excised and homogenized in PBS. Levels of IFN-β (B), IFN-λ3 (C), and IL-18 (D) in intestine homogenates were quantified by ELISA.(E and F) The wild-type Dhx15fl/fl and Dhx15IEC-KO mice (n = 15 per strain) were inoculated intragastrically with 1 × 108 PFU of reovirus T3D strain. At day 4 post-inoculation, mice were euthanized, feces were collected, and intestinal tissues were excised. The viral titers in intestine homogenates (E) and shedding in feces (F) were determined by plaque assay. Results are expressed as mean viral titers for 15 animals for each time point. Error bars indicate SEMs.(G) Flow cytometry analysis of CD4+ T cells, CD8+ T cells, B cells, and NK cells of intestine lamina propria lymphocytes (left panel) and mesenteric lymph nodes (mLN) (right panel) from wild-type Dhx15fl/fl and Dhx15IEC-KO mice infected with reovirus for 2 days using CD3-FITC, CD4-PE/cyanine7, CD8a-PerCP/cyanine5.5, CD19-APC, and NK1.1-PE antibodies.(H) The absolute cell numbers in intestine (left) and mLN (right) from wild-type Dhx15fl/fl and Dhx15IEC-KO mice (n = 3 mice) for representative flow cytometry data in (G).(I) Hematoxylin and eosin (H&E) staining of intestine sections from wild-type Dhx15fl/fl and Dhx15IEC-KO mice as in (E). Scale bars represent 200 μm.(J) Graph depicting histology scores for inflammation and tissue damage of intestine sections in (I). Data are represented as means ± SEMs. NS, p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 (unpaired t test).

Fig 3: Anti-CD8 treatment potentiates the PC progression. (A) Six-week-old male C57BL/6 mice were intraperitoneally inoculated with MC-38 cells and then treated with anti-CD8 antibody or IgG as a control. The tumor nodes in visceral fats of each mouse were observed on day 15. Representative images were displayed. (B) The weight of tumor nodes in visceral fats as described above in (A). The data represent the means±s.e.ms. (n=3, ***P<0.005, Student's t-test).

Fig 4: Flow cytometry analysis of memory CD8 T and CD4 T cells and total percentages of CD4 T and CD8 T cells in spleen and blood samples at 36 days after rechallenge. A Percentages of CD4 and CD8 memory T cells in spleen. B Percentages of CD4 and CD8 memory T cells in blood. C Percentages of CD4/CD45 and CD8/CD45 in spleen. D Percentages of CD4/CD45 and CD8/CD45 in blood. Data are expressed as means ± standard error of the mean (SEM). P < 0.05 was considered statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS not significant. HDXRT high-dose radiotherapy; LDXRT low-dose radiotherapy; ICIs immune checkpoint inhibitors

Fig 5: BoxA increases survival and induces immunization in a syngeneic mouse model of mesothelioma AScheme of the experiment. BALB/c mice were inoculated i.p. with 7 × 104 MM cells and treated with either 800 µg BoxA (red bars) or PBS (blue bars) three times a week, 10 times in total. Yellow arrows represent BLI imaging.BTreatment with BoxA at increasing doses reduced the number of mice with detectable tumor lesions in a statistically significant dose‐dependent manner (Spearman correlation: P = 0.04). In this experiment, no BLI measurement was taken.CForty mice were inoculated with MM cells and treated or not with BoxA. Tumor growth was detected via BLI. Lines that do not reach day 34 correspond to mice that were sacrificed for ethical reasons.DKaplan–Meier survival curves. Statistics: log‐rank Gehan–Breslow–Wilcoxon test, P < 0.0001, n = 20 per group.ETumor growth after MM re‐challenge detected by BLI.F–GFlow cytometry analysis of luciferase‐specific T cells. In (F), representative dot plots depict intracellular IFNγ levels in gated CD8+ CD44high T cells. In (G), differences in the percentage of IFNγ producing CD8+ CD44high cells, pulsed relative to unpulsed. Statistics: Kruskal–Wallis test; each dot represents a mouse (n = 3–7 per group); bars represent mean ± SD. Source data are available online for this figure.