Fig 2: Decrease of myeloid cells during anti-angiogenic therapy. (A) The total number of monocytes and neutrophils (F) was determined by automatic hematologic cell count before (T1) and after the first cycle (T2) and at the end of anti-angiogenic therapy (T3). The percentage of CD14+ within CD45+ cells (B) and the percentage of VEGF-R1+ (C) and Tie2+ (D) and VEGFR-1+Tie2+ (E) within CD14+ cells were determined by cytometry before (T1), after the first cycle of anti-angiogenic therapy (T2), and at the end of anti-angiogenic therapy (T3). Statistical analysis was performed with GraphPad Prism® software using the paired t-test.

Fig 3: Sunitinib modulates pro-angiogenic monocytes in murine tumors. Balb/c mice were grafted subcutaneously with CT-26 tumor cells (0.2 × 105 cells). When the tumors reached 9–10 mm2, mice were treated with sunitinib by oral gavage at 40 mg/kg daily or dimethylsulfoxide (DMSO) 10% in PBS. Mice were sacrificed at D20 post tumor graft. Analyses of Tie2+ VEGFR-1+ CD11b myeloid cells by flow cytometry in blood (A,B) and in tumors (C,D) are shown. Triangles and circles correspond to mice treated by DMSO and sunitinib respectively. Data are representative of two independent experiments with 5 mice/group. Data are mean ±SEM. Difference between the two groups were analyzed using with the Mann-whitney t-test. ** p < 0.01.

Fig 4: Characterization of human pro-angiogenic monocytes in blood. Peripheral blood mononuclear cells were stained with anti-CD45, anti-CD14, anti-VEGFR-1 and anti-Tie 2. The data analysis was performed as follows: Selection of CD45+ cells then live cells followed by gating on CD14+ cells and expression of VEGFR-1 or Tie-2+ cells on the monocyte population. Isotype controls corresponding to anti-VEGR-1 (IgG1 PE) and anti-Tie2 (IgG1 APC) wereshown (bottom right). Data acquisition was performed with a Navios (Beckman Coulter) and analyses were performed with Kaluza software (Beckman Coulter, Villepinte 95942 Roissy CDG, France).