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Intravitreal TSG-6 suppresses laser-induced choroidal neovascularization by inhibiting CCR2+ monocyte recruitment.

Kim SJ, Lee HJ, Yun JH, Ko JH, Choi da Y, Oh JY - Sci Rep (2015)

Bottom Line: An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV.Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes.Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea [2] Samsung Biomedical Research Institute, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea.

ABSTRACT
Choroidal neovascularization (CNV) is the hallmark of wet age-related macular degeneration (AMD), one of the leading causes of blindness in the elderly. Although the pathogenesis of CNV is not clear, a number of studies show that ocular-infiltrating macrophages and inflammation play a critical role in the development of CNV. TNFα-stimulated gene/protein (TSG)-6 is a multifunctional endogenous protein that has anti-inflammatory activities partly by regulating macrophage activation. Therefore, we here investigated the therapeutic potential of TSG-6 in a rat model of CNV induced by laser photocoagulation. Time course analysis showed that the expression of VEGF and pro-inflammatory cytokines in the choroid was up-regulated early after laser injury, and gradually decreased to baseline over 14 days. An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV. Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes. Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment. Together, the results demonstrate that TSG-6 inhibits inflammation and CCR2(+) monocyte recruitment into the choroid, and suppresses the development of CNV.

No MeSH data available.


Related in: MedlinePlus

TSG-6 inhibits CCR2+ monocyte recruitment to CNV lesion.(a) Immunohistochemical staining of the RPE-choroid-scleral flat mounts at day 3 showed a massive infiltration of CCR2+ cells in the area of CNV after injury. There was significantly less infiltration of CCR2+ cells in TSG-6-treated eyes. (b) Similarly, Iba+ cells largely infiltrated the CNV lesion, and TSG-6 treatment significantly reduced Iba+ cell infiltration. Original magnification × 200. Scale bar: 100 μm. Photographs shown are representative of two independent experiments (each with four eyes per group), and data are presented as mean + SEM. *p < 0.05.
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f4: TSG-6 inhibits CCR2+ monocyte recruitment to CNV lesion.(a) Immunohistochemical staining of the RPE-choroid-scleral flat mounts at day 3 showed a massive infiltration of CCR2+ cells in the area of CNV after injury. There was significantly less infiltration of CCR2+ cells in TSG-6-treated eyes. (b) Similarly, Iba+ cells largely infiltrated the CNV lesion, and TSG-6 treatment significantly reduced Iba+ cell infiltration. Original magnification × 200. Scale bar: 100 μm. Photographs shown are representative of two independent experiments (each with four eyes per group), and data are presented as mean + SEM. *p < 0.05.

Mentions: In addition, we performed histological assays to localize CCR2+ monocytes infiltrating the choroid upon laser injury. Immunohistochemical staining of the RPE-choroid-scleral flat mounts revealed large infiltration of CCR2+ cells in the CNV lesion at day 3 after injury (Fig. 4a). Consistent with flow cytometric results, there were significantly fewer CCR2+ cells in TSG-6-treated eyes (Fig. 4a). Further assay showed that a large number of Iba1+ cells indicating microglia and monocytes were accumulated in the lesion of CNV (Fig. 4b), reflecting the activation of microglia and infiltration of monocytes. TSG-6 treatment significantly decreased the number of Iba1+ cells in the injury site (Fig. 4b).


Intravitreal TSG-6 suppresses laser-induced choroidal neovascularization by inhibiting CCR2+ monocyte recruitment.

Kim SJ, Lee HJ, Yun JH, Ko JH, Choi da Y, Oh JY - Sci Rep (2015)

TSG-6 inhibits CCR2+ monocyte recruitment to CNV lesion.(a) Immunohistochemical staining of the RPE-choroid-scleral flat mounts at day 3 showed a massive infiltration of CCR2+ cells in the area of CNV after injury. There was significantly less infiltration of CCR2+ cells in TSG-6-treated eyes. (b) Similarly, Iba+ cells largely infiltrated the CNV lesion, and TSG-6 treatment significantly reduced Iba+ cell infiltration. Original magnification × 200. Scale bar: 100 μm. Photographs shown are representative of two independent experiments (each with four eyes per group), and data are presented as mean + SEM. *p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493567&req=5

f4: TSG-6 inhibits CCR2+ monocyte recruitment to CNV lesion.(a) Immunohistochemical staining of the RPE-choroid-scleral flat mounts at day 3 showed a massive infiltration of CCR2+ cells in the area of CNV after injury. There was significantly less infiltration of CCR2+ cells in TSG-6-treated eyes. (b) Similarly, Iba+ cells largely infiltrated the CNV lesion, and TSG-6 treatment significantly reduced Iba+ cell infiltration. Original magnification × 200. Scale bar: 100 μm. Photographs shown are representative of two independent experiments (each with four eyes per group), and data are presented as mean + SEM. *p < 0.05.
Mentions: In addition, we performed histological assays to localize CCR2+ monocytes infiltrating the choroid upon laser injury. Immunohistochemical staining of the RPE-choroid-scleral flat mounts revealed large infiltration of CCR2+ cells in the CNV lesion at day 3 after injury (Fig. 4a). Consistent with flow cytometric results, there were significantly fewer CCR2+ cells in TSG-6-treated eyes (Fig. 4a). Further assay showed that a large number of Iba1+ cells indicating microglia and monocytes were accumulated in the lesion of CNV (Fig. 4b), reflecting the activation of microglia and infiltration of monocytes. TSG-6 treatment significantly decreased the number of Iba1+ cells in the injury site (Fig. 4b).

Bottom Line: An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV.Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes.Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea [2] Samsung Biomedical Research Institute, 50 Irwon-dong, Gangnam-gu, Seoul, 135-710, Korea.

ABSTRACT
Choroidal neovascularization (CNV) is the hallmark of wet age-related macular degeneration (AMD), one of the leading causes of blindness in the elderly. Although the pathogenesis of CNV is not clear, a number of studies show that ocular-infiltrating macrophages and inflammation play a critical role in the development of CNV. TNFα-stimulated gene/protein (TSG)-6 is a multifunctional endogenous protein that has anti-inflammatory activities partly by regulating macrophage activation. Therefore, we here investigated the therapeutic potential of TSG-6 in a rat model of CNV induced by laser photocoagulation. Time course analysis showed that the expression of VEGF and pro-inflammatory cytokines in the choroid was up-regulated early after laser injury, and gradually decreased to baseline over 14 days. An intravitreal injection of TSG-6 suppressed the expression of VEGF and pro-inflammatory cytokines including CCL2, and reduced the size of CNV. Also, the number of Iba(+) and CCR2(+) cells including infiltrating macrophages was markedly lower in the CNV lesion of TSG-6-treated eyes. Further analysis identified CCR2(+) CD11b(+) CD11c(+) cells and CCR2(+) CD11b(-)CD11c(+) cells as the cell populations that were increased by laser injury and reduced by TSG-6 treatment. Together, the results demonstrate that TSG-6 inhibits inflammation and CCR2(+) monocyte recruitment into the choroid, and suppresses the development of CNV.

No MeSH data available.


Related in: MedlinePlus