Expression and differential regulation of HLA-G isoforms in the retinal pigment epithelial cell line, ARPE-19

Signe Goul Svendsen; Maja Søberg Udsen; Marina Daouya; Tina Funck; Ching-Lien Wu; Edgardo D. Carosella; Joel LeMaoult; Thomas Vauvert F. Hviid; Carsten Faber; Mogens Holst Nissen

doi:10.1016/j.humimm.2017.04.007

Expression and differential regulation of HLA-G isoforms in the retinal pigment epithelial cell line, ARPE-19

Signe Goul Svendsen^*, Maja Søberg Udsen, Marina Daouya, Tina Funck, Ching-Lien Wu, Edgardo D. Carosella, Joel LeMaoult, Thomas Vauvert F. Hviid, Carsten Faber, Mogens Holst Nissen

^*Corresponding author for this work

7 Citations (Scopus)

Abstract

The purpose of this study was to examine if HLA-G is expressed in the retinal pigment epithelium (RPE) cells of the eye. The RPE comprises the outer most layer of the retina and as such defines the interface to the blood and contributes to the immune privilege in the posterior part of the eye. One way the RPE might be regulating the immune system could be by expressing the non-classical human leukocyte antigen (HLA) molecule, HLA-G. We therefore sought to define if the RPE cell line, ARPE-19, expressed HLA-G and analyse the regulation as a response to pro-inflammatory cytokines. This was done by digital droplet PCR, measuring the gene expression of HLA-G in total RNA. The protein expression was analysed by immunohistochemistry and by immunofluorescence followed by confocal microscopy and the expression of the HLA-G isoforms was explored by fragment analysis. In the current study, we show that HLA-G is expressed by ARPE-19 cells and is upregulated as a response to pro-inflammatory cytokines. Moreover, we are the first to describe a differential regulation of the HLA-G isoforms as a direct response to stimulation. These results might indicate that HLA-G can be part of the immune privilege of the posterior part of the eye, but further experiments on primary RPE cells are needed.

Original language	English
Journal	Human Immunology
Volume	78
Issue number	5-6
Pages (from-to)	414-420
Number of pages	7
ISSN	0198-8859
DOIs	https://doi.org/10.1016/j.humimm.2017.04.007
Publication status	Published - 1 May 2017

Keywords

ARPE-19
HLA-G
Isoforms
Retinal pigment epithelium
sHLA-G

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10.1016/j.humimm.2017.04.007

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title = "Expression and differential regulation of HLA-G isoforms in the retinal pigment epithelial cell line, ARPE-19",

abstract = "The purpose of this study was to examine if HLA-G is expressed in the retinal pigment epithelium (RPE) cells of the eye. The RPE comprises the outer most layer of the retina and as such defines the interface to the blood and contributes to the immune privilege in the posterior part of the eye. One way the RPE might be regulating the immune system could be by expressing the non-classical human leukocyte antigen (HLA) molecule, HLA-G. We therefore sought to define if the RPE cell line, ARPE-19, expressed HLA-G and analyse the regulation as a response to pro-inflammatory cytokines. This was done by digital droplet PCR, measuring the gene expression of HLA-G in total RNA. The protein expression was analysed by immunohistochemistry and by immunofluorescence followed by confocal microscopy and the expression of the HLA-G isoforms was explored by fragment analysis. In the current study, we show that HLA-G is expressed by ARPE-19 cells and is upregulated as a response to pro-inflammatory cytokines. Moreover, we are the first to describe a differential regulation of the HLA-G isoforms as a direct response to stimulation. These results might indicate that HLA-G can be part of the immune privilege of the posterior part of the eye, but further experiments on primary RPE cells are needed.",

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author = "Svendsen, {Signe Goul} and Udsen, {Maja S{\o}berg} and Marina Daouya and Tina Funck and Ching-Lien Wu and Carosella, {Edgardo D.} and Joel LeMaoult and Hviid, {Thomas Vauvert F.} and Carsten Faber and Nissen, {Mogens Holst}",

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AU - Svendsen, Signe Goul

AU - Udsen, Maja Søberg

AU - Daouya, Marina

AU - Funck, Tina

AU - Wu, Ching-Lien

AU - Carosella, Edgardo D.

AU - LeMaoult, Joel

AU - Hviid, Thomas Vauvert F.

AU - Faber, Carsten

AU - Nissen, Mogens Holst

PY - 2017/5/1

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N2 - The purpose of this study was to examine if HLA-G is expressed in the retinal pigment epithelium (RPE) cells of the eye. The RPE comprises the outer most layer of the retina and as such defines the interface to the blood and contributes to the immune privilege in the posterior part of the eye. One way the RPE might be regulating the immune system could be by expressing the non-classical human leukocyte antigen (HLA) molecule, HLA-G. We therefore sought to define if the RPE cell line, ARPE-19, expressed HLA-G and analyse the regulation as a response to pro-inflammatory cytokines. This was done by digital droplet PCR, measuring the gene expression of HLA-G in total RNA. The protein expression was analysed by immunohistochemistry and by immunofluorescence followed by confocal microscopy and the expression of the HLA-G isoforms was explored by fragment analysis. In the current study, we show that HLA-G is expressed by ARPE-19 cells and is upregulated as a response to pro-inflammatory cytokines. Moreover, we are the first to describe a differential regulation of the HLA-G isoforms as a direct response to stimulation. These results might indicate that HLA-G can be part of the immune privilege of the posterior part of the eye, but further experiments on primary RPE cells are needed.

AB - The purpose of this study was to examine if HLA-G is expressed in the retinal pigment epithelium (RPE) cells of the eye. The RPE comprises the outer most layer of the retina and as such defines the interface to the blood and contributes to the immune privilege in the posterior part of the eye. One way the RPE might be regulating the immune system could be by expressing the non-classical human leukocyte antigen (HLA) molecule, HLA-G. We therefore sought to define if the RPE cell line, ARPE-19, expressed HLA-G and analyse the regulation as a response to pro-inflammatory cytokines. This was done by digital droplet PCR, measuring the gene expression of HLA-G in total RNA. The protein expression was analysed by immunohistochemistry and by immunofluorescence followed by confocal microscopy and the expression of the HLA-G isoforms was explored by fragment analysis. In the current study, we show that HLA-G is expressed by ARPE-19 cells and is upregulated as a response to pro-inflammatory cytokines. Moreover, we are the first to describe a differential regulation of the HLA-G isoforms as a direct response to stimulation. These results might indicate that HLA-G can be part of the immune privilege of the posterior part of the eye, but further experiments on primary RPE cells are needed.

KW - ARPE-19

KW - HLA-G

KW - Isoforms

KW - Retinal pigment epithelium

KW - sHLA-G

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DO - 10.1016/j.humimm.2017.04.007

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IS - 5-6

ER -

Expression and differential regulation of HLA-G isoforms in the retinal pigment epithelial cell line, ARPE-19

Abstract

Keywords

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