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. 2019 Apr 17;8(8):1601482. doi: 10.1080/2162402X.2019.1601482

Figure 4.

Figure 4.

Treatment with a P4D2 mAb shifts primary human monocyte differentiation away from protumor phenotype. Human primary monocytes were differentiated with human AB serum in presence of P4D2 or P1D9 mAbs. Two other conditions included: (1) human serum and (2) human stable galectin-9 (hG9NC). (a) Representative figures from the flow cytometric analysis of CD68+ and CCR5+ mature macrophages. (b) Percentages of CD68+ and CCR5+ cells are showed for the different treatments. Statistical differences among treatments (n = 3/group) were assessed with one-way ANOVA followed by the Bonferroni test and indicated with *P ≤ 0.01. (c) Illustrative schematic of monocyte-macrophage differentiation experiment. (d) Monocyte-macrophage differentiation was evaluated using supernatants from ROB MM cells (conditioned media) or from ROB MM cells treated with P4D2 mAb (conditioned media + P4D2). P4D2-treated cells were used as control. Flow cytometry representative images show the reduction of CD68+ and CCR5+ mature macrophages induced by P4D2-treated ROB media. (e) Differences in the percentage of CD68+ and CCR5+ cells in ROB media compared to P4D2 mAb-treated ROB media, or P4D2 (n = 3/group) were evaluated using one-way ANOVA followed by the Bonferroni test and indicated with *P ≤ 0.001. (f) Maturation of primary monocytes was also measured with real-time PCR using primers for the M2 marker, MARCO. Differences in ROB media compared to P4D2 mAb-treated ROB media for MARCO were defined using one-tailed paired Student’s t test (n = 3/group) and indicated with *P ≤ 0.001.