Lactoferrin Expression in Human and Murine Ocular Tissue

Abrar A. Rageh, Deborah A. Ferrington, Heidi Roehrich, Ching Yuan, Marcia R. Terluk, Elizabeth F. Nelson, Sandra R. Montezuma

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Purpose: Lactoferrin (LF) is a multifunctional protein known to provide innate defense due to its antimicrobial and anti-inflammatory properties. In the eye, LF has been identified in the tears and vitreous humor. Its presence in other ocular tissues has not been determined. Our aim is to assess the presence of LF in the cornea, iris, retina and retinal pigment epithelium (RPE) of humans and mice. Methods: To test for the endogenous production of LF, reverse transcription polymerase chain reaction was performed in cultured human cells from the cornea and RPE and in murine tissues. To confirm LF localization in specific ocular tissue, immunohistochemistry was performed on flat mounts of cornea, retina and RPE in human donor eyes. The presence of LF was assessed by western blotting in human and mouse ocular tissue and human culture cells (cornea and RPE). To verify antibody specificity, purified human LF and transferrin (TF) were used on 1D and 2D western blots. Results: LF gene expression was confirmed in the cornea and RPE cell cultures from humans, suggesting that LF is an endogenously produced protein. PCR results from mouse ocular tissue showed LF expression in cornea, iris, RPE, but not in retina. These results were also consistent with immunohistochemical localization of LF in human donor tissue. Antibody reaction for human LF was specific and western blotting showed its presence in the cornea, iris and RPE tissues. A faint reaction for the retina was observed but was likely due to contamination from other ocular tissues. Multiple commercially available antibodies for murine LF cross-reacted with TF, so no reliable results were obtained for murine western blot. Conclusion: LF is expressed in multiple eye tissues of humans and mice. This widespread expression and multifunctional activity of LF suggests that it may play an important role in protecting eye tissues from inflammation-associated diseases.

Original languageEnglish (US)
Pages (from-to)883-889
Number of pages7
JournalCurrent Eye Research
Volume41
Issue number7
DOIs
StatePublished - Jul 2 2016

Fingerprint

Lactoferrin
Retinal Pigment Epithelium
Cornea
Retina
Iris
Western Blotting
Transferrin
Cell Culture Techniques
Vitreous Body
Polymerase Chain Reaction
Antibody Specificity
Antibodies
Tears
Reverse Transcription

Keywords

  • Cornea
  • iris
  • lactoferrin
  • retina
  • retinal pigment epithelium
  • transferrin

Cite this

Lactoferrin Expression in Human and Murine Ocular Tissue. / Rageh, Abrar A.; Ferrington, Deborah A.; Roehrich, Heidi; Yuan, Ching; Terluk, Marcia R.; Nelson, Elizabeth F.; Montezuma, Sandra R.

In: Current Eye Research, Vol. 41, No. 7, 02.07.2016, p. 883-889.

Research output: Contribution to journalArticle

Rageh, Abrar A. ; Ferrington, Deborah A. ; Roehrich, Heidi ; Yuan, Ching ; Terluk, Marcia R. ; Nelson, Elizabeth F. ; Montezuma, Sandra R. / Lactoferrin Expression in Human and Murine Ocular Tissue. In: Current Eye Research. 2016 ; Vol. 41, No. 7. pp. 883-889.
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abstract = "Purpose: Lactoferrin (LF) is a multifunctional protein known to provide innate defense due to its antimicrobial and anti-inflammatory properties. In the eye, LF has been identified in the tears and vitreous humor. Its presence in other ocular tissues has not been determined. Our aim is to assess the presence of LF in the cornea, iris, retina and retinal pigment epithelium (RPE) of humans and mice. Methods: To test for the endogenous production of LF, reverse transcription polymerase chain reaction was performed in cultured human cells from the cornea and RPE and in murine tissues. To confirm LF localization in specific ocular tissue, immunohistochemistry was performed on flat mounts of cornea, retina and RPE in human donor eyes. The presence of LF was assessed by western blotting in human and mouse ocular tissue and human culture cells (cornea and RPE). To verify antibody specificity, purified human LF and transferrin (TF) were used on 1D and 2D western blots. Results: LF gene expression was confirmed in the cornea and RPE cell cultures from humans, suggesting that LF is an endogenously produced protein. PCR results from mouse ocular tissue showed LF expression in cornea, iris, RPE, but not in retina. These results were also consistent with immunohistochemical localization of LF in human donor tissue. Antibody reaction for human LF was specific and western blotting showed its presence in the cornea, iris and RPE tissues. A faint reaction for the retina was observed but was likely due to contamination from other ocular tissues. Multiple commercially available antibodies for murine LF cross-reacted with TF, so no reliable results were obtained for murine western blot. Conclusion: LF is expressed in multiple eye tissues of humans and mice. This widespread expression and multifunctional activity of LF suggests that it may play an important role in protecting eye tissues from inflammation-associated diseases.",
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AU - Ferrington, Deborah A.

AU - Roehrich, Heidi

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AU - Terluk, Marcia R.

AU - Nelson, Elizabeth F.

AU - Montezuma, Sandra R.

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