The present study investigated the interactions among the complement membrane attack complex (MAC), CCL2, and VEGF that occur in vivo during the development of choroidal neovascularization (CNV). We first investigated the sequential expression of MAC, CCL2, and VEGF during laser-induced CNV in C57BL/6 mice. Increased MAC deposition was detected at 1 h, CCL2 increased at 3 h, and VEGF was up-regulated at day 3 post-laser treatment. These results suggested that during laser-induced CNV, MAC, CCL2 and VEGF are formed and/or expressed in the following order: MAC 3 CCL2 3 VEGF. To determine the cross-talk between MAC, CCL2, and VEGF during laser-induced CNV, neutralizing antibodies were injected both systemically and locally to block the bioactivity of each molecule. Blocking MAC formation inhibited CCL2 and VEGF expression and also limited CNV formation, whereas neutralization of CCL2 bioactivity did not affect MAC deposition; however, it reduced VEGF expression and CNV formation. When bioactivity of VEGF was blocked, CNV formation was significantly inhibited, but MAC deposition was not affected. Together, our results demonstrate that MAC is an upstream mediator and effect of MAC on the development of laser-induced CNV can be attributed to its direct effect on VEGF as well as its effect on VEGF that is mediated by CCL2. Understanding the interplay between immune mediators is critical to gain insight into the pathogenesis of CNV.
Age-related macular degeneration (AMD)3 is the major cause of irreversible blindness in individuals over the age of 50 worldwide (1-5). Based on clinical and pathological features, AMD is usually classified into two forms: the nonexudative/dry form and the exudative/wet form (6, 7). Wet AMD is a pathological process, secondary to choroidal neovascular changes (CNV). CNV is a complex pathogenic process where new blood vessels are generated beneath the retina from pre-existing choriocapillaries (8 -10). Several risk factors have been reported to be associated with CNV formation (11-15). Studies reported in the literature support a key role for the complement system in the development of CNV (16 -21).Although the complement system is recognized traditionally as a major component of innate immunity, it has multifunctional role in immunity, including the initiation and regulation of adaptive immune responses (22-26). The role of the complement in the development of CNV has been more directly addressed by using laser-induced animal model (27-29). The laser-induced model of CNV is produced in C57BL/6 mice by laser photocoagulation, and this model is used by an increasing number of investigators (29 -32). Using this animal model of CNV, we demonstrated that complement activation, especially the formation of membrane attack complex (MAC), is crucial for the development of laser-induced CNV (27-29). We also reported that there is a direct correlation between MAC deposition and levels of angiogenic growth factors, including VEGF during laser-induced CNV (29).Studies reported in the literature have indi...