Fig 1: Absolute counts for expression of CD4, CD8, CD68 and IL-2 molecules in treated and untreated tumors.P values were calculated using Mann-Whitney test for comparison of absolute cell counts between untreated (TM) and treated (TM-F3) tumor-bearing mice.
Fig 2: The contribution of ozone exposure to airway smooth function and Th1/Th2 balance of asthmatic mice. (A–E) The bronchial wall thickness (A), smooth muscle thickness (B), airway resistance (C), lung compliance (D) and Log PC100 Penh (E) of mice was examined among saline+air, saline+ozone, OVA+air and OVA+ozone groups. *: P<0.05 when compared with saline+air group, #: P<0.05 when compared with saline+ozone group, &: P<0.05 when compared with OVA+air group. (F–H) Levels of HA (F), TNF-α (G) and IL-13 (H) were determined within mice of saline+air, saline+ozone, OVA+air and OVA+ozone groups. *: P<0.05 when compared with saline+air group, #: P<0.05 when compared with saline+ozone group, &: P<0.05 when compared with OVA+air group. (I–O) The amounts of IL-5 (I), IL-4 (J), IL-10 (K), IFN-γ (L), IL-2 (M), T-bet (N) and GATA3 (O) were determined within mice treated by saline+air, saline+ozone, OVA+air and OVA+ozone. *: P<0.05 when compared with saline+air group, #: P<0.05 when compared with saline+ozone group, &: P<0.05 when compared with OVA+air group.
Fig 3: a. Photographs of foot pad thickness (white arrow) observed in animals of different treatment groups. Scale bar = 5 mm; b. Graphical representation of foot pad thickness (mm) in different groups. Data expressed as mean ± S.D.; (one way ANOVA, n = 5 in each group); ∗(P < 0.05), ∗∗(P < 0.01), ∗∗∗(P < 0.001). c. Shows increase in antibody titer in extract (150 mg/kg, 300 mg/kg) treated mice compared to the saline treated. d. Shows the stimulatory effect of A. reticulata extract (150 mg/kg & 300 mg/kg) on antibody titer in immunosuppressed (CP treated) mice. Data expressed as mean rank difference ± SEM (Kruskal–Wallis test, n = 5 in each group). e. Dot plots (FACS) representing intracellular IL-2 and IL-6 expression in subpopulation of PBMC of different treatment groups. The upper panel shows the CD3e positive cells (lymphocytes) expressing IL-2 within the whole PBMC population; the lower panel shows the CD14 positive cells (monocytes & macrophages) expressing IL-6. Data from one representative experiment out of three are shown. Numbers in the upper right (UR) quadrants show percentages of positive cells expressing IL. The mean of that percentage value in different groups from 3 different experiments was plotted graphically to evaluate the difference between groups (left panels). (One way ANOVA; n = 3); ∗(P < 0.05), ∗∗(P < 0.01), ∗∗∗(P < 0.001); ns (P ˃ 0.05); PE = phycoerythrin, FITC = fluorescein isothiocyanate; CP = Cyclophosphamide; Ex150 = Extract 150 mg/kg; Ex300 = Extract 300 mg/kg.
Fig 4: Profile of interleukin expression in spleen tissue in different treatment groups. Immunostaining was performed with antibody against IL-2 and IL-6 in mouse spleen tissue. (a) Shows the immunoreactivity of spleen tissue against IL-2 and (b) shows the same against IL-6. In all the panels, immunoreactivity can be observed as brown deposits (black arrow). Magnification 400×; Scale bar = 100 μm. Each image in a single panel represents different treatment groups. CP = Cyclophosphamide; Ex150 = Extract 150 mg/kg; Ex300 = Extract 300 mg/kg.
Fig 5: Gut microbiota changes the expression of immune factors in the tumor microenvironment. (A) Bar graph of PD-1, TGF-β, IFN-γ, IL-2, IL-6, and IL-17 in by Elisa tumor samples. (B) The number of CD4+ and CD8+ T cell were detected by flow cytometry. (C) Representative immunohistochemistry images on tumor samples.
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