Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells.

Alessandra Micera, Barbara Stampachiacchiere, Eduardo Maria Normando, Alessandro Lambiase, Sergio Bonini, Stefano Bonini

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

PURPOSE: To investigate if nerve growth factor (NGF) might modulate toll-like receptor (TLR) 4 and 9 expression in primary cultures of vernal keratoconjunctivitis (VKC)-derived conjunctival epithelial cells (VKC-ECs). METHODS: Primary cultures of ECs were established from VKC (n=7) and healthy-control (n=5) conjunctival specimens. Primary untouched and short-term NGF-exposed VKC-ECs were analyzed for molecular (relative real-time PCR) and biochemical (confocal and fluorescence-activated cell sorting analysis of TLR4 and TLR9 expression. Data were compared to their untreated as well as stimulated healthy-control counterparts. Conditioned media were analyzed for interferon (IFN)-gamma, interleukin (IL)-4, IL-10, and IL-12 p40 secretion. RESULTS: Primary untouched VKC-ECs showed TLR4 increase and TLR9 decrease compared to their healthy-control counterparts. NGF exposure resulted in a strong upregulation of TLR4 and a moderate upregulation of TLR9 in few passages VKC-ECs. Both TLR4 and TLR9 upregulation occurred in a dose-dependent fashion and were supported by biochemical analysis. NGF triggered a dose-response release of IL-10 in VKC-ECs conditioned media, an effect not detected for IL-4, IL-12 p40, and IFN-gamma. CONCLUSIONS: Our data indicate that NGF is able to induce TLR4/TLR9 overexpression in VKC-ECs. These cells exhibited poor IL-4, IL-12 p40, and IFN-gamma responses to NGF, while a significant IL-10 decreased secretion was detected. The different NGF-induced TLR response between VKC and healthy-control conjunctival ECs as well as the different cytokine response might reflect a different pattern of cell activation according to the state of VKC.

Original languageEnglish
Pages (from-to)2037-2044
Number of pages8
JournalMolecular Vision
Volume15
Publication statusPublished - 2009

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Toll-Like Receptor 9
Allergic Conjunctivitis
Toll-Like Receptor 4
Nerve Growth Factor
Epithelial Cells
Interleukin-12
Interleukin-4
Interleukin-10
Interferon-gamma
Up-Regulation
Conditioned Culture Medium
Toll-Like Receptors
Real-Time Polymerase Chain Reaction
Flow Cytometry
Cytokines

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Micera, A., Stampachiacchiere, B., Normando, E. M., Lambiase, A., Bonini, S., & Bonini, S. (2009). Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells. Molecular Vision, 15, 2037-2044.

Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells. / Micera, Alessandra; Stampachiacchiere, Barbara; Normando, Eduardo Maria; Lambiase, Alessandro; Bonini, Sergio; Bonini, Stefano.

In: Molecular Vision, Vol. 15, 2009, p. 2037-2044.

Research output: Contribution to journalArticle

Micera, A, Stampachiacchiere, B, Normando, EM, Lambiase, A, Bonini, S & Bonini, S 2009, 'Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells.', Molecular Vision, vol. 15, pp. 2037-2044.
Micera, Alessandra ; Stampachiacchiere, Barbara ; Normando, Eduardo Maria ; Lambiase, Alessandro ; Bonini, Sergio ; Bonini, Stefano. / Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells. In: Molecular Vision. 2009 ; Vol. 15. pp. 2037-2044.
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T1 - Nerve growth factor modulates toll-like receptor (TLR) 4 and 9 expression in cultured primary VKC conjunctival epithelial cells.

AU - Micera, Alessandra

AU - Stampachiacchiere, Barbara

AU - Normando, Eduardo Maria

AU - Lambiase, Alessandro

AU - Bonini, Sergio

AU - Bonini, Stefano

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N2 - PURPOSE: To investigate if nerve growth factor (NGF) might modulate toll-like receptor (TLR) 4 and 9 expression in primary cultures of vernal keratoconjunctivitis (VKC)-derived conjunctival epithelial cells (VKC-ECs). METHODS: Primary cultures of ECs were established from VKC (n=7) and healthy-control (n=5) conjunctival specimens. Primary untouched and short-term NGF-exposed VKC-ECs were analyzed for molecular (relative real-time PCR) and biochemical (confocal and fluorescence-activated cell sorting analysis of TLR4 and TLR9 expression. Data were compared to their untreated as well as stimulated healthy-control counterparts. Conditioned media were analyzed for interferon (IFN)-gamma, interleukin (IL)-4, IL-10, and IL-12 p40 secretion. RESULTS: Primary untouched VKC-ECs showed TLR4 increase and TLR9 decrease compared to their healthy-control counterparts. NGF exposure resulted in a strong upregulation of TLR4 and a moderate upregulation of TLR9 in few passages VKC-ECs. Both TLR4 and TLR9 upregulation occurred in a dose-dependent fashion and were supported by biochemical analysis. NGF triggered a dose-response release of IL-10 in VKC-ECs conditioned media, an effect not detected for IL-4, IL-12 p40, and IFN-gamma. CONCLUSIONS: Our data indicate that NGF is able to induce TLR4/TLR9 overexpression in VKC-ECs. These cells exhibited poor IL-4, IL-12 p40, and IFN-gamma responses to NGF, while a significant IL-10 decreased secretion was detected. The different NGF-induced TLR response between VKC and healthy-control conjunctival ECs as well as the different cytokine response might reflect a different pattern of cell activation according to the state of VKC.

AB - PURPOSE: To investigate if nerve growth factor (NGF) might modulate toll-like receptor (TLR) 4 and 9 expression in primary cultures of vernal keratoconjunctivitis (VKC)-derived conjunctival epithelial cells (VKC-ECs). METHODS: Primary cultures of ECs were established from VKC (n=7) and healthy-control (n=5) conjunctival specimens. Primary untouched and short-term NGF-exposed VKC-ECs were analyzed for molecular (relative real-time PCR) and biochemical (confocal and fluorescence-activated cell sorting analysis of TLR4 and TLR9 expression. Data were compared to their untreated as well as stimulated healthy-control counterparts. Conditioned media were analyzed for interferon (IFN)-gamma, interleukin (IL)-4, IL-10, and IL-12 p40 secretion. RESULTS: Primary untouched VKC-ECs showed TLR4 increase and TLR9 decrease compared to their healthy-control counterparts. NGF exposure resulted in a strong upregulation of TLR4 and a moderate upregulation of TLR9 in few passages VKC-ECs. Both TLR4 and TLR9 upregulation occurred in a dose-dependent fashion and were supported by biochemical analysis. NGF triggered a dose-response release of IL-10 in VKC-ECs conditioned media, an effect not detected for IL-4, IL-12 p40, and IFN-gamma. CONCLUSIONS: Our data indicate that NGF is able to induce TLR4/TLR9 overexpression in VKC-ECs. These cells exhibited poor IL-4, IL-12 p40, and IFN-gamma responses to NGF, while a significant IL-10 decreased secretion was detected. The different NGF-induced TLR response between VKC and healthy-control conjunctival ECs as well as the different cytokine response might reflect a different pattern of cell activation according to the state of VKC.

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