Fig 2: Cannabidiol (CBD) suppresses alternative activation of macrophage in vitro. (A) Immunofluorescence staining of tumor tissue from vehicle or CBD treatment mice with CD206 and inducible NO synthase (iNOS). (B–D) Bone marrow-derived macrophages (BMDMs) were stimulated with media or interleukin-4 (IL-4) (20 ng/mL) then treated with dimethyl sulfoxide (DMSO) (vehicle control) or increasing concentrations (μM) of CBD for 24 h. (B) Flow cytometry data for the M2-specific surface marker CD206 (left) and M1 surface marker CD86 (right). (C) Quantitative real-time polymerase chain reaction (qRT-PCR) results showing mRNA expression of Arg1, Cd206, Chil3, and Retnla normalized to β-actin. (D) qRT-PCR results showing mRNA expression of Il12b and Nos2 normalized to β-actin. (E, F) BMDMs were stimulated with medium or IL-4 and treated with DMSO or CBD for 24 h. Immunofluorescence images showing the expression of CD206 (E) and iNOS (F). (G) BMDMs were stimulated with medium or IL-4 (20 ng/mL) and treated with DMSO or CBD for 24 h then co-cultured with MC38-mCheery cells for another 48 h. (H) Time course images of organoid on day 1 and day 4 (n = 6–8 for each time point). Data are represented as mean ± standard error of mean. ∗P < 0.05 and ∗∗P < 0.01. ns: no significance.

Fig 3: Effects of doxorubicin on macrophage activation. A, The experimental design of the study. B, Flow cytometry histograms of circulating monocytes (CD45+CD11b+Ly6G−Ly6C+), showing Ly6Chi monocytes (R1) and Ly6Clo monocytes (R2). The proportion of Ly6Chi and Ly6Clo monocytes to the population of circulating monocytes (C and D). E, Flow cytometry dot plots displaying M2 macrophages (CD45+CD11b+F4/80+CD206+, R1) in hearts. F, Number of total macrophages (CD45+CD11b+F4/80+) per mg of tissue. G, Percentage of macrophages based on the population of CD45+ leukocytes in the heart. Number of M1 (CD45+CD11b+F4/80+CD86+) and M2 macrophages per mg of tissue, and percentage of M1 and M2 macrophages to the population of macrophages (H through K). Immunofluorescence staining showing M1 macrophages visualized by red fluorescence (CD86+) and M2 macrophages visualized by green fluorescence (CD206+) (L). Number of CD86+ and CD206+ macrophages per view (M and N). Statistical analysis was performed by 1‐way ANOVA. *P<0.033, **P<0.002, ***P<0.001, n=6/group. CTL indicates control; and DOX, doxorubicin.

Fig 4: Percent of CD14+ monocytes of total leukocytes (live CD45+) (Paired Mann‐Whitney U test; p = 0.0039) (A) and activated monocytes (HLA‐DR+CD86+) of total monocytes (live CD45+CD14+) (Paired Mann‐Whitney U test; p = 0.002) (B) were significantly higher in peripheral blood than in placental bed samples

Fig 5: Term uterine leukocytes contained a reduced proportion of B cells compared to peripheral blood leukocytes. Uterine blood mononuclear cells (UBMC) contained significantly lowers percentages of CD19+ B cells when compared to that of PBMC (paired t‐test; p = 0.0052) (A); Activated (HLA‐DR+CD86+) percentages of B cells were not significantly different in PBMC and UBMC (Paired Mann‐Whitney U test; p = 0.92) (B)