Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro

Marina Mesquida; Faye Drawnel; Philippa J Lait; David A Copland; Madeleine L Stimpson; Victor Llorenç; Maite Sainz de la Maza; Alfredo Adan; Gabriella Widmer; Pamela Strassburger; Sascha Fauser; Andrew D Dick; Richard W J Lee; Blanca Molins

doi:10.1167/tvst.8.5.32

Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro

Marina Mesquida, Faye Drawnel, Philippa J Lait, David A Copland, Madeleine L Stimpson, Victor Llorenç, Maite Sainz de la Maza, Alfredo Adan, Gabriella Widmer, Pamela Strassburger, Sascha Fauser, Andrew D Dick, Richard W J Lee, Blanca Molins

Research output: Contribution to journal › Article (Academic Journal) › peer-review

44 Citations (Scopus)

206 Downloads (Pure)

Abstract

Purpose: Macular edema (ME) is a leading cause of visual loss in a range of retinal diseases and despite the use of antivascular endothelial growth factor (anti-VEGF) agents, its successful treatment remains a major clinical challenge. Based on the indirect clinical evidence that interleukin-6 (IL-6) is a key additional candidate mediator of ME, we interrogated the effect of IL-6 on blood-retinal barrier (BRB) integrity in vitro.

Methods: Human retinal pigment epithelial cell (ARPE-19) and human retinal microvascular endothelial cell (HRMEC) monolayers were used to mimic the outer and inner BRB, respectively. Their paracellular permeability was assessed by measuring the passive permeation of 40 kDa fluorescein isothiocyanate (FITC)-dextran across confluent cells in the presence of IL-6. Transendothelial/epithelial electrical resistance (TEER) then was measured and the distribution of the tight junction protein ZO-1 was assessed by immunofluorescence using confocal microscopy.

Results: Treatment with IL-6 for 48 hours significantly increased the diffusion rate of FITC-dextran, decreased TEER, and disrupted the distribution of ZO-1 in ARPE-19 cells, which constitutively express the IL-6 transmembrane receptor, and this was reversed with IL-6R blockade. In contrast, IL-6 did not affect the paracellular permeability, TEER, or ZO-1 distribution in HRMECs.

Conclusions: These in vitro data support the hypothesis that IL-6 reversibly disrupts the integrity of ARPE-19 cells, but it does not affect HRMECs.

Translational Relevance: IL-6 is a candidate therapeutic target in the treatment of outer BRB driven ME.

Original language	English
Article number	32
Number of pages	10
Journal	Translational vision science & technology
Volume	8
Issue number	5
DOIs	https://doi.org/10.1167/tvst.8.5.32
Publication status	Published - 28 Oct 2019

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Access to Document

10.1167/tvst.8.5.32

Full-text PDF (final published version)
This is the final published version of the article (version of record). It first appeared online via Association for Research in Vision and Ophthalmology at https://tvst.arvojournals.org/article.aspx?articleid=2753751&resultClick=1 Please refer to any applicable terms of use of the publisher.
Final published version, 2.43 MBLicence: CC BY

Handle.net

Persistent link

Cite this

Mesquida, M., Drawnel, F., Lait, P. J., Copland, D. A., Stimpson, M. L., Llorenç, V., Sainz de la Maza, M., Adan, A., Widmer, G., Strassburger, P., Fauser, S., Dick, A. D., Lee, R. W. J., & Molins, B. (2019). Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro. Translational vision science & technology, 8(5), Article 32. https://doi.org/10.1167/tvst.8.5.32

@article{d445b93d18304d62aeff6e8b096c3bdc,

title = "Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro",

abstract = "Purpose: Macular edema (ME) is a leading cause of visual loss in a range of retinal diseases and despite the use of antivascular endothelial growth factor (anti-VEGF) agents, its successful treatment remains a major clinical challenge. Based on the indirect clinical evidence that interleukin-6 (IL-6) is a key additional candidate mediator of ME, we interrogated the effect of IL-6 on blood-retinal barrier (BRB) integrity in vitro.Methods: Human retinal pigment epithelial cell (ARPE-19) and human retinal microvascular endothelial cell (HRMEC) monolayers were used to mimic the outer and inner BRB, respectively. Their paracellular permeability was assessed by measuring the passive permeation of 40 kDa fluorescein isothiocyanate (FITC)-dextran across confluent cells in the presence of IL-6. Transendothelial/epithelial electrical resistance (TEER) then was measured and the distribution of the tight junction protein ZO-1 was assessed by immunofluorescence using confocal microscopy.Results: Treatment with IL-6 for 48 hours significantly increased the diffusion rate of FITC-dextran, decreased TEER, and disrupted the distribution of ZO-1 in ARPE-19 cells, which constitutively express the IL-6 transmembrane receptor, and this was reversed with IL-6R blockade. In contrast, IL-6 did not affect the paracellular permeability, TEER, or ZO-1 distribution in HRMECs.Conclusions: These in vitro data support the hypothesis that IL-6 reversibly disrupts the integrity of ARPE-19 cells, but it does not affect HRMECs.Translational Relevance: IL-6 is a candidate therapeutic target in the treatment of outer BRB driven ME.",

author = "Marina Mesquida and Faye Drawnel and Lait, \{Philippa J\} and Copland, \{David A\} and Stimpson, \{Madeleine L\} and Victor Lloren{\c c} and \{Sainz de la Maza\}, Maite and Alfredo Adan and Gabriella Widmer and Pamela Strassburger and Sascha Fauser and Dick, \{Andrew D\} and Lee, \{Richard W J\} and Blanca Molins",

year = "2019",

month = oct,

day = "28",

doi = "10.1167/tvst.8.5.32",

language = "English",

volume = "8",

journal = "Translational vision science \& technology",

issn = "2164-2591",

publisher = "Association for Research in Vision and Ophthalmology",

number = "5",

}

Mesquida, M, Drawnel, F, Lait, PJ , Copland, DA, Stimpson, ML, Llorenç, V, Sainz de la Maza, M, Adan, A, Widmer, G, Strassburger, P, Fauser, S, Dick, AD, Lee, RWJ & Molins, B 2019, 'Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro', Translational vision science & technology, vol. 8, no. 5, 32. https://doi.org/10.1167/tvst.8.5.32

TY - JOUR

T1 - Modelling Macular Edema

T2 - The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro

AU - Mesquida, Marina

AU - Drawnel, Faye

AU - Lait, Philippa J

AU - Copland, David A

AU - Stimpson, Madeleine L

AU - Llorenç, Victor

AU - Sainz de la Maza, Maite

AU - Adan, Alfredo

AU - Widmer, Gabriella

AU - Strassburger, Pamela

AU - Fauser, Sascha

AU - Dick, Andrew D

AU - Lee, Richard W J

AU - Molins, Blanca

PY - 2019/10/28

Y1 - 2019/10/28

N2 - Purpose: Macular edema (ME) is a leading cause of visual loss in a range of retinal diseases and despite the use of antivascular endothelial growth factor (anti-VEGF) agents, its successful treatment remains a major clinical challenge. Based on the indirect clinical evidence that interleukin-6 (IL-6) is a key additional candidate mediator of ME, we interrogated the effect of IL-6 on blood-retinal barrier (BRB) integrity in vitro.Methods: Human retinal pigment epithelial cell (ARPE-19) and human retinal microvascular endothelial cell (HRMEC) monolayers were used to mimic the outer and inner BRB, respectively. Their paracellular permeability was assessed by measuring the passive permeation of 40 kDa fluorescein isothiocyanate (FITC)-dextran across confluent cells in the presence of IL-6. Transendothelial/epithelial electrical resistance (TEER) then was measured and the distribution of the tight junction protein ZO-1 was assessed by immunofluorescence using confocal microscopy.Results: Treatment with IL-6 for 48 hours significantly increased the diffusion rate of FITC-dextran, decreased TEER, and disrupted the distribution of ZO-1 in ARPE-19 cells, which constitutively express the IL-6 transmembrane receptor, and this was reversed with IL-6R blockade. In contrast, IL-6 did not affect the paracellular permeability, TEER, or ZO-1 distribution in HRMECs.Conclusions: These in vitro data support the hypothesis that IL-6 reversibly disrupts the integrity of ARPE-19 cells, but it does not affect HRMECs.Translational Relevance: IL-6 is a candidate therapeutic target in the treatment of outer BRB driven ME.

AB - Purpose: Macular edema (ME) is a leading cause of visual loss in a range of retinal diseases and despite the use of antivascular endothelial growth factor (anti-VEGF) agents, its successful treatment remains a major clinical challenge. Based on the indirect clinical evidence that interleukin-6 (IL-6) is a key additional candidate mediator of ME, we interrogated the effect of IL-6 on blood-retinal barrier (BRB) integrity in vitro.Methods: Human retinal pigment epithelial cell (ARPE-19) and human retinal microvascular endothelial cell (HRMEC) monolayers were used to mimic the outer and inner BRB, respectively. Their paracellular permeability was assessed by measuring the passive permeation of 40 kDa fluorescein isothiocyanate (FITC)-dextran across confluent cells in the presence of IL-6. Transendothelial/epithelial electrical resistance (TEER) then was measured and the distribution of the tight junction protein ZO-1 was assessed by immunofluorescence using confocal microscopy.Results: Treatment with IL-6 for 48 hours significantly increased the diffusion rate of FITC-dextran, decreased TEER, and disrupted the distribution of ZO-1 in ARPE-19 cells, which constitutively express the IL-6 transmembrane receptor, and this was reversed with IL-6R blockade. In contrast, IL-6 did not affect the paracellular permeability, TEER, or ZO-1 distribution in HRMECs.Conclusions: These in vitro data support the hypothesis that IL-6 reversibly disrupts the integrity of ARPE-19 cells, but it does not affect HRMECs.Translational Relevance: IL-6 is a candidate therapeutic target in the treatment of outer BRB driven ME.

U2 - 10.1167/tvst.8.5.32

DO - 10.1167/tvst.8.5.32

M3 - Article (Academic Journal)

C2 - 31667008

SN - 2164-2591

VL - 8

JO - Translational vision science & technology

JF - Translational vision science & technology

IS - 5

M1 - 32

ER -

Modelling Macular Edema: The Effect of IL-6 and IL-6R Blockade on Human Blood-Retinal Barrier Integrity In Vitro

Abstract

UN SDGs

Access to Document

Handle.net

Fingerprint

Cite this