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Abrar A. Rageh, Marcia R. Terluk, Ching Yuan, Deborah A. Ferrington, Sandra R. Montezuma; Lactoferrin in the Retina and Retinal Pigment Epithelium of Humans and Mice. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3223.
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Lactoferrin (LF) is a protein identified in secretions from exocrine glands and in specific granules of neutrophils. It is known to provide innate defense due to its anti-microbial and anti-inflammatory properties. In the human eye, LF is present in tears and has been identified in the vitreous. Its presence in other eye tissue has not been determined. Our aim is to assess whether LF is present in the retina and Retinal Pigment Epithelium (RPE), which is a site of inflammation associated with retinal diseases, such as uveitis and Age-related Macular Degeneration.
C57BL/6J mice were sacrificed and perfused with phosphate buffered saline. The eyes were enucleated and the RPE and retinal tissues were isolated and processed for protein. Human retina and RPE cells were harvested from donor eyes obtained from the Minnesota Lions Eye Bank. The presence of LF was assessed by Western Blotting using LF anti-rabbit antibody. Bovine LF was used as a positive control and murine duodenum was used as a negative control. To confirm the endogenous production of LF, RPE from mouse or human donors were cultured in 4% or 15% Fetal Bovine Serum, respectively. To generate stress, cells were switched to Serum Free Media and harvested 2 and 6 hours later.
Western blot analysis of human and mouse tissue revealed the presence of LF in the retina and RPE tissues. LF expression was confirmed in the RPE cell cultures from humans and mice, suggesting that LF is an endogenous protein. After 2 and 6 hours of stressing the RPE cells under serum-free conditions, the LF signal was increased in the media compared to the media from non-stressed cells.
Our findings show that LF is present in the retina and RPE of humans and mice. This protein is also secreted by the RPE under stress conditions. Considering the anti-microbial and anti-inflammatory properties of LF, our results suggest LF may play an important role in protecting the retina from infectious agents and inflammation associated with disease.
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