The University of Southampton
University of Southampton Institutional Repository

Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression

Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression
Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression
Background and Aims: Previous studies suggest that low serum 25-hydroxyvitamin D [25(OH) D] levels are associated with reduced responsiveness to interferon and ribavirin therapy. We investigated the impact of vitamin D metabolites on HCV and cellular gene expression in cultured hepatoma cells.

Methods: HCV Replicon cell lines stably expressing luciferase reporter constructs (genotype 1b and 2a replicon) or JC1-Luc2a were incubated in the presence of vitamin D2, vitamin D3 or 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Presence of HCV was quantified by a luciferase reporter assay and immunoblot of the Core protein. Synergy of interferon-alpha A/D (IFN-α) and 1,25(OH)2D3 was evaluated using the Chou-Talalay method. Cellular gene expression by microarray analysis using Illumina Bead Chips and real-time quantitative PCR.

Results: Vitamin D2, D3 and 1,25(OH)2D3 each demonstrated anti- HCV activity at low micro molar concentrations. In vitro conversion from D3 to 25(OH)D3 was shown by LC/MS/MS. Combination indices of 1,25(OH)2D3 and IFN-α demonstrated a synergistic effect (0.23-0.46) and significantly reduced core expression by immunoblot. Differentially expressed genes were identified between Huh7.5.1 cells in the presence and absence of 1,25(OH)2D3 and HCV. Genes involved with classical effects of vitamin D metabolism and excretion were activated, along with genes linked to autophagy such as G-protein coupled receptor 37 (GPR37) and Hypoxiainducible factor 1-alpha (HIF1a). Additionally, additive effects of 1,25(OH)2D3 and IFN-α were seen on mRNA expression of chemokine motif ligand 20 (CCL20).

Conclusions: This study shows that vitamin D reduces HCV protein production in cell culture synergistically with IFN-α. Vitamin D also activates gene expression independently and additively with IFN-α and this may explain its ability to aid in the clearance of HCV in vivo.
Gutierrez, J.A.
c8bea97c-504e-4baa-b9a1-73451f558165
Jones, K.A.
2333d02e-c72b-4420-9b36-a554915b8e3d
Flores, R.
580e477b-714c-45eb-93f8-f77bc3fe41ba
Singhania, A.
9a5f2c6b-fc46-4223-bcae-de70bf14da6b
Woelk, C.H.
4d3af0fd-658f-4626-b3b5-49a6192bcf7d
Schooley, R.T.
a5fa4402-5e04-4f70-8a8d-92035524ff12
Wyles, D.L
c777d43c-1f22-40e3-81e0-803b76a8583b
Gutierrez, J.A.
c8bea97c-504e-4baa-b9a1-73451f558165
Jones, K.A.
2333d02e-c72b-4420-9b36-a554915b8e3d
Flores, R.
580e477b-714c-45eb-93f8-f77bc3fe41ba
Singhania, A.
9a5f2c6b-fc46-4223-bcae-de70bf14da6b
Woelk, C.H.
4d3af0fd-658f-4626-b3b5-49a6192bcf7d
Schooley, R.T.
a5fa4402-5e04-4f70-8a8d-92035524ff12
Wyles, D.L
c777d43c-1f22-40e3-81e0-803b76a8583b

Gutierrez, J.A., Jones, K.A., Flores, R., Singhania, A., Woelk, C.H., Schooley, R.T. and Wyles, D.L (2014) Vitamin D metabolites inhibit hepatitis C virus and modulate cellular gene expression. Journal of Virology & Antiviral Research, 3 (3). (doi:10.4172/2324-8955.1000129).

Record type: Article

Abstract

Background and Aims: Previous studies suggest that low serum 25-hydroxyvitamin D [25(OH) D] levels are associated with reduced responsiveness to interferon and ribavirin therapy. We investigated the impact of vitamin D metabolites on HCV and cellular gene expression in cultured hepatoma cells.

Methods: HCV Replicon cell lines stably expressing luciferase reporter constructs (genotype 1b and 2a replicon) or JC1-Luc2a were incubated in the presence of vitamin D2, vitamin D3 or 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Presence of HCV was quantified by a luciferase reporter assay and immunoblot of the Core protein. Synergy of interferon-alpha A/D (IFN-α) and 1,25(OH)2D3 was evaluated using the Chou-Talalay method. Cellular gene expression by microarray analysis using Illumina Bead Chips and real-time quantitative PCR.

Results: Vitamin D2, D3 and 1,25(OH)2D3 each demonstrated anti- HCV activity at low micro molar concentrations. In vitro conversion from D3 to 25(OH)D3 was shown by LC/MS/MS. Combination indices of 1,25(OH)2D3 and IFN-α demonstrated a synergistic effect (0.23-0.46) and significantly reduced core expression by immunoblot. Differentially expressed genes were identified between Huh7.5.1 cells in the presence and absence of 1,25(OH)2D3 and HCV. Genes involved with classical effects of vitamin D metabolism and excretion were activated, along with genes linked to autophagy such as G-protein coupled receptor 37 (GPR37) and Hypoxiainducible factor 1-alpha (HIF1a). Additionally, additive effects of 1,25(OH)2D3 and IFN-α were seen on mRNA expression of chemokine motif ligand 20 (CCL20).

Conclusions: This study shows that vitamin D reduces HCV protein production in cell culture synergistically with IFN-α. Vitamin D also activates gene expression independently and additively with IFN-α and this may explain its ability to aid in the clearance of HCV in vivo.

This record has no associated files available for download.

More information

Accepted/In Press date: 3 October 2014
e-pub ahead of print date: 2014
Published date: 6 October 2014
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 379212
URI: http://eprints.soton.ac.uk/id/eprint/379212
PURE UUID: 03b0c956-cc30-4271-b185-079b7b665c4a

Catalogue record

Date deposited: 18 Jul 2015 14:32
Last modified: 14 Mar 2024 20:35

Export record

Altmetrics

Contributors

Author: J.A. Gutierrez
Author: K.A. Jones
Author: R. Flores
Author: A. Singhania
Author: C.H. Woelk
Author: R.T. Schooley
Author: D.L Wyles

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×