Infiltration of COX-2–expressing macrophages is a prerequisite for IL-1β–induced neovascularization and tumor growth
J. Clin. Invest. Shintaro Nakao, et al. 115:2979 doi:10.1172/JCI23298 [Go to this article.]

Figure 3
The role of MCP-1 in IL-1β– or VEGF-induced angiogenesis. (A) Kinetics of MCP-1 levels after pellet implantation. Corneal lysates were prepared and assayed by ELISA at the indicated times (n = 3). *P < 0.01 and **P < 0.03 versus untreated (N). (B) Kinetics of infiltrating macrophages in IL-1β–implanted corneas. Corneal lysates were prepared from untreated and IL-1β–treated corneas on the days shown (n = 3). Percentages of infiltrating F4/80⁺ cells were quantified using FACS. (C) Corneal neovascularization induced by IL-1β (30 ng) or VEGF (200 ng) in C57BL/6 wild-type and MCP-1^–/– mice on day 6. (D) Corneal neovascularization at the indicated time points. (E) Quantitative analysis of IL-1β–induced corneal neovascularization in MCP-1^–/– (n = 10) and wild-type mice (n = 8) on day 6. Bars show means ± SD. *P < 0.01 versus wild-type mice using the unpaired Student’s t test. (F) Immunohistochemistry for Gr-1 or F4/80 (brown) in corneal sections on day 6 after IL-1β pellet implantation in MCP-1^–/– or wild-type mice. Gr-1–positive cells were detected in both types of mice. F4/80-positive cells were detected on day 6 in wild-type but not MCP-1^–/– mice. (G) FACS analysis of infiltrating cells from 5 IL-1β– or VEGF-implanted corneas at day 6 in wild-type or MCP-1^–/– mice. The percentages of CD11b⁺F4/80⁺ cells were 4.09% ± 2.13% (IL-1β, wild-type), 2.53% ± 1.73% (IL-1β, MCP-1^–/–), 0.30% ± 0.10% (VEGF, wild-type), and 0.14% ± 0.05% (VEGF, MCP-1^–/–). The percentages of CD11b⁺Gr-1⁺ cells were 13.5% ± 2.89% (IL-1β, wild-type), 7.84% ± 0.48% (IL-1β, MCP-1^–/–), 0.94% ± 0.55% (VEGF, wild-type) and 0.20% ± 0.10% (VEGF, MCP-1^–/–).