Expression of human beta-defensins in intraocular tissues

R J Haynes, J E McElveen, H S Dua, P J Tighe, J Liversidge

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

PURPOSE. Defensins are naturally occurring antimicrobial peptides. Recently the authors published evidence of defensin production by the human ocular surface. A study was undertaken to look for intraocular defensins that may account for unexplained antimicrobial activity of intraocular fluids.

METHODS. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on human postmortem ciliary body samples for beta defensins-1 (HBD-1) and beta defensin-2 (HBD-2), and alpha defensins 5 and 6. Induction of defensins by cytokines was analyzed in cultured human ciliary body epithelial (CBE) and retinal pigment epithelial (RPE) cells. Polyclonal antibodies were used to immunoblot aqueous and vitreous to detect HBD-1 and HBD-2 and to estimate their concentration.

RESULTS. RT-PCR revealed constitutive HBD-1 message in ciliary body. HBD-2 and alpha defensin 5 and 6 messages were absent. HBD-2 message was induced by cytokine stimulation of both CBE and RPE cells. Immunoblots of vitreous and aqueous stained positively for HBD-1 but not HBD-2. The estimated aqueous concentration of HBD-1 was less than 16 ng/ml.

CONCLUSIONS. This Study demonstrates that HBD-1 is constitutively present in the aqueous and vitreous. probably at sub-bacteriocidal concentrations. HBD-2, was absent from aqueous, but cytokine stimulation studies suggest that it may be generated in response to inflammatory cytokines during infections. HBD-2 has a wider antibacterial spectrum, is 10-fold more potent, and may play a more significant role in antimicrobial defense than HBD-1. The use of defensins therapeutically may be indicated; however, caution is required because defensins also promote cell proliferation and fibrin formation, which are 2 key elements in ocular scarring processes such as proliferative vitreoretinopathy.

Original languageEnglish
Pages (from-to)3026-3031
Number of pages6
JournalInvestigative Ophthalmology & Visual Science
Volume41
Publication statusPublished - 2000

Keywords

  • ANTIMICROBIAL PEPTIDE GENE
  • EPITHELIAL-CELLS
  • PROLIFERATIVE VITREORETINOPATHY
  • CATARACT-SURGERY
  • MESSENGER-RNA
  • BACTERIAL-CONTAMINATION
  • PLASMINOGEN-ACTIVATOR
  • HUMAN BETA-DEFENSIN-2
  • HUMAN-NEUTROPHILS
  • AQUEOUS-HUMOR

Cite this

Haynes, R. J., McElveen, J. E., Dua, H. S., Tighe, P. J., & Liversidge, J. (2000). Expression of human beta-defensins in intraocular tissues. Investigative Ophthalmology & Visual Science, 41, 3026-3031.

Expression of human beta-defensins in intraocular tissues. / Haynes, R J ; McElveen, J E ; Dua, H S ; Tighe, P J ; Liversidge, J .

In: Investigative Ophthalmology & Visual Science, Vol. 41, 2000, p. 3026-3031.

Research output: Contribution to journalArticle

Haynes, RJ, McElveen, JE, Dua, HS, Tighe, PJ & Liversidge, J 2000, 'Expression of human beta-defensins in intraocular tissues', Investigative Ophthalmology & Visual Science, vol. 41, pp. 3026-3031.
Haynes RJ, McElveen JE, Dua HS, Tighe PJ, Liversidge J. Expression of human beta-defensins in intraocular tissues. Investigative Ophthalmology & Visual Science. 2000;41:3026-3031.
Haynes, R J ; McElveen, J E ; Dua, H S ; Tighe, P J ; Liversidge, J . / Expression of human beta-defensins in intraocular tissues. In: Investigative Ophthalmology & Visual Science. 2000 ; Vol. 41. pp. 3026-3031.
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TY - JOUR

T1 - Expression of human beta-defensins in intraocular tissues

AU - Haynes, R J

AU - McElveen, J E

AU - Dua, H S

AU - Tighe, P J

AU - Liversidge, J

PY - 2000

Y1 - 2000

N2 - PURPOSE. Defensins are naturally occurring antimicrobial peptides. Recently the authors published evidence of defensin production by the human ocular surface. A study was undertaken to look for intraocular defensins that may account for unexplained antimicrobial activity of intraocular fluids.METHODS. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on human postmortem ciliary body samples for beta defensins-1 (HBD-1) and beta defensin-2 (HBD-2), and alpha defensins 5 and 6. Induction of defensins by cytokines was analyzed in cultured human ciliary body epithelial (CBE) and retinal pigment epithelial (RPE) cells. Polyclonal antibodies were used to immunoblot aqueous and vitreous to detect HBD-1 and HBD-2 and to estimate their concentration.RESULTS. RT-PCR revealed constitutive HBD-1 message in ciliary body. HBD-2 and alpha defensin 5 and 6 messages were absent. HBD-2 message was induced by cytokine stimulation of both CBE and RPE cells. Immunoblots of vitreous and aqueous stained positively for HBD-1 but not HBD-2. The estimated aqueous concentration of HBD-1 was less than 16 ng/ml.CONCLUSIONS. This Study demonstrates that HBD-1 is constitutively present in the aqueous and vitreous. probably at sub-bacteriocidal concentrations. HBD-2, was absent from aqueous, but cytokine stimulation studies suggest that it may be generated in response to inflammatory cytokines during infections. HBD-2 has a wider antibacterial spectrum, is 10-fold more potent, and may play a more significant role in antimicrobial defense than HBD-1. The use of defensins therapeutically may be indicated; however, caution is required because defensins also promote cell proliferation and fibrin formation, which are 2 key elements in ocular scarring processes such as proliferative vitreoretinopathy.

AB - PURPOSE. Defensins are naturally occurring antimicrobial peptides. Recently the authors published evidence of defensin production by the human ocular surface. A study was undertaken to look for intraocular defensins that may account for unexplained antimicrobial activity of intraocular fluids.METHODS. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on human postmortem ciliary body samples for beta defensins-1 (HBD-1) and beta defensin-2 (HBD-2), and alpha defensins 5 and 6. Induction of defensins by cytokines was analyzed in cultured human ciliary body epithelial (CBE) and retinal pigment epithelial (RPE) cells. Polyclonal antibodies were used to immunoblot aqueous and vitreous to detect HBD-1 and HBD-2 and to estimate their concentration.RESULTS. RT-PCR revealed constitutive HBD-1 message in ciliary body. HBD-2 and alpha defensin 5 and 6 messages were absent. HBD-2 message was induced by cytokine stimulation of both CBE and RPE cells. Immunoblots of vitreous and aqueous stained positively for HBD-1 but not HBD-2. The estimated aqueous concentration of HBD-1 was less than 16 ng/ml.CONCLUSIONS. This Study demonstrates that HBD-1 is constitutively present in the aqueous and vitreous. probably at sub-bacteriocidal concentrations. HBD-2, was absent from aqueous, but cytokine stimulation studies suggest that it may be generated in response to inflammatory cytokines during infections. HBD-2 has a wider antibacterial spectrum, is 10-fold more potent, and may play a more significant role in antimicrobial defense than HBD-1. The use of defensins therapeutically may be indicated; however, caution is required because defensins also promote cell proliferation and fibrin formation, which are 2 key elements in ocular scarring processes such as proliferative vitreoretinopathy.

KW - ANTIMICROBIAL PEPTIDE GENE

KW - EPITHELIAL-CELLS

KW - PROLIFERATIVE VITREORETINOPATHY

KW - CATARACT-SURGERY

KW - MESSENGER-RNA

KW - BACTERIAL-CONTAMINATION

KW - PLASMINOGEN-ACTIVATOR

KW - HUMAN BETA-DEFENSIN-2

KW - HUMAN-NEUTROPHILS

KW - AQUEOUS-HUMOR

M3 - Article

VL - 41

SP - 3026

EP - 3031

JO - Investigative Ophthalmology & Visual Science

JF - Investigative Ophthalmology & Visual Science

SN - 0146-0404

ER -