PURPOSE: To investigate the role of neutrophils in the development of laser induced experimental choroidal neovascularization (CNV). METHODS: CNV was induced by laser photocoagulation in adult male C57BL/6J mice. Neutrophil infiltration was evaluated by histology and confocal immunohistology. The expression of neutrophil chemotactic chemokines in the regions of laser injury was determined by quantitative real-time PCR. Animals were treated with NIMP-R14, an anti-murine neutrophil monoclonal antibody (mAb), intraperitoneally to deplete neutrophils. The specific neutrophil depletion was confirmed by flow cytometry. The CNV responses were compared between neutropenic and untreated control mice on the basis of fluorescein angiography (FA), CNV lesion volume and lesion histology, and vascular endothelial growth factor (VEGF) expression by ELISA. Expression of VEGF and Angiopoietin-1 and Angiopoietin-2 protein by murine neutrophils was evaluated by confocal immunohistochemistry. RESULTS: Neutrophils infiltrated the sites of laser injury as early as day 1 after laser treatment and peaked at day 3. The neutrophil infiltration correlated with enhanced mRNA expression of neutrophil chemotactic chemokines MIP-2 and KC in the lesions. Administration of NIMP-R14 mAb specifically depleted neutrophils. Analysis of FA, CNV volume, and lesion histology, all demonstrated a moderate decrease in the CNV response in neutropenic mice compared to control mice (p<0.01). The reduction in the CNV response in neutropenic mice was associated with decreased VEGF protein levels in the ocular posterior segment. Murine neutrophils contained VEGF and Angiopoietin-1 and Angiopoietin-2 proteins. CONCLUSIONS: Neutrophil invasion was part of early inflammatory responses during laser induced CNV. Neutrophil depletion correlated with reduced CNV responses and decreased VEGF protein expression. These data suggest that neutrophils promoted the early development of CNV possibly via secretion of angiogenic growth factors.
PURPOSE: To investigate the role of neutrophils in the development of laser induced experimental choroidal neovascularization (CNV). METHODS: CNV was induced by laser photocoagulation in adult male C57BL/6J mice. Neutrophil infiltration was evaluated by histology and confocal immunohistology. The expression of neutrophil chemotactic chemokines in the regions of laser injury was determined by quantitative real-time PCR. Animals were treated with NIMP-R14, an anti-murine neutrophil monoclonal antibody (mAb), intraperitoneally to deplete neutrophils. The specific neutrophil depletion was confirmed by flow cytometry. The CNV responses were compared between neutropenic and untreated control mice on the basis of fluorescein angiography (FA), CNV lesion volume and lesion histology, and vascular endothelial growth factor (VEGF) expression by ELISA. Expression of VEGF and Angiopoietin-1 and Angiopoietin-2 protein by murine neutrophils was evaluated by confocal immunohistochemistry. RESULTS: Neutrophils infiltrated the sites of laser injury as early as day 1 after laser treatment and peaked at day 3. The neutrophil infiltration correlated with enhanced mRNA expression of neutrophil chemotactic chemokines MIP-2 and KC in the lesions. Administration of NIMP-R14 mAb specifically depleted neutrophils. Analysis of FA, CNV volume, and lesion histology, all demonstrated a moderate decrease in the CNV response in neutropenicmice compared to control mice (p<0.01). The reduction in the CNV response in neutropenicmice was associated with decreased VEGF protein levels in the ocular posterior segment. Murine neutrophils contained VEGF and Angiopoietin-1 and Angiopoietin-2 proteins. CONCLUSIONS: Neutrophil invasion was part of early inflammatory responses during laser induced CNV. Neutrophil depletion correlated with reduced CNV responses and decreased VEGF protein expression. These data suggest that neutrophils promoted the early development of CNV possibly via secretion of angiogenic growth factors.
Authors: Yijun Hu; Haijiang Lin; Bernard Dib; Alp Atik; Peggy Bouzika; Christopher Lin; Yueran Yan; Shibo Tang; Joan W Miller; Demetrios G Vavvas Journal: Discov Med Date: 2014 Jul-Aug Impact factor: 2.970
Authors: Kristopher G Sheets; Yongdong Zhou; Monica K Ertel; Eric J Knott; Cornelius E Regan; Jasmine R Elison; William C Gordon; Per Gjorstrup; Nicolas G Bazan Journal: Mol Vis Date: 2010-03-02 Impact factor: 2.367
Authors: Atsunobu Takeda; Judit Z Baffi; Mark E Kleinman; Won Gil Cho; Miho Nozaki; Kiyoshi Yamada; Hiroki Kaneko; Romulo J C Albuquerque; Sami Dridi; Kuniharu Saito; Brian J Raisler; Steven J Budd; Pete Geisen; Ariel Munitz; Balamurali K Ambati; Martha G Green; Tatsuro Ishibashi; John D Wright; Alison A Humbles; Craig J Gerard; Yuichiro Ogura; Yuzhen Pan; Justine R Smith; Salvatore Grisanti; M Elizabeth Hartnett; Marc E Rothenberg; Jayakrishna Ambati Journal: Nature Date: 2009-06-14 Impact factor: 49.962