Regulation of T-lymphocyte CCL3 and CCL4 production by retinal pigment epithelial cells

Carol A. Wallace; Gill Moir; David F. G. Malone; Linda Duncan; Gayathri Devarajan; Isabel J. Crane

doi:10.1167/iovs.12-10602

Regulation of T-lymphocyte CCL3 and CCL4 production by retinal pigment epithelial cells

Carol A. Wallace, Gill Moir, David F. G. Malone, Linda Duncan, Gayathri Devarajan, Isabel J. Crane^*

^*Corresponding author for this work

University of Aberdeen

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

PURPOSE. Retinal pigment epithelial (RPE) cells have an important role in the immune suppression associated with the immune privilege of the eye. Some aspects of this remain unclear and this study aimed to determine how RPE cells could influence the production of chemokines by T lymphocytes.

METHODS. T lymphocytes, separated from peripheral blood of normal volunteers, and RPE cells, cultured from donor eyes, were cultured separately and together, either in contact or in transwells. Supernatants were analyzed for CCL3, CCL4, and soluble CD54 (sCD54) by ELISA. Blocking agents were used to determine which soluble mediators were involved.

RESULTS. Coculture of RPE cells with activated lymphocytes resulted in a reduction in CCL3 and CCL4 production by lymphocytes, primarily by soluble mediators. Soluble CD54 was markedly increased on coculture of lymphocytes with RPE cells. Soluble CD54 reduced CCL3 and CCL4 production by RPE cells, and inhibition of CCL3 and CCL4 on coculture with RPE cells was reduced by anti-CD54. Blocking prostaglandin E2 (PGE2) abrogated the inhibition of CCL4, but not CCL3, by RPE cells. Blocking TGF beta and nitric oxide production had no effect.

CONCLUSIONS. RPE cells are able to down-regulate high levels of CCL3 and CCL4 production by T lymphocytes by using the soluble mediators sCD54 and PGE2. Reducing this production of CCL3 and CCL4 will dampen down the cascade effect and recruitment of more inflammatory cells, protecting the retina from an excessive immune response. (Invest Ophthalmol Vis Sci. 2013; 54: 722-730) DOI:10.1167/iovs.12-10602

Original language	English
Pages (from-to)	722-730
Number of pages	9
Journal	Investigative Ophthalmology & Visual Science
Volume	54
Issue number	1
Early online date	8 Jan 2013
DOIs	https://doi.org/10.1167/iovs.12-10602
Publication status	Published - 28 Jan 2013

Keywords

intercellular-adhesion molecule-1
nitric-oxide synthase
experimental autoimmune enecephalomyelitis
function-associated antigen-1
gene-expression
inflammatory protein-1-alpha
endothelial-cells
immune privilege
beta-chemokines
soluble ICAM-1

Access to Document

10.1167/iovs.12-10602

Iain Fraser Cytometry Centre
Andrea Holme (Manager), Linda Duncan (Senior Application Scientist), Ailsa Laird (Technician) & Kate Burgoyne (Technician)
Institute of Medical Sciences
Research Facilities: Facility

Cite this

@article{f41d0911f14c4ecabef25feb894cc672,

title = "Regulation of T-lymphocyte CCL3 and CCL4 production by retinal pigment epithelial cells",

abstract = "PURPOSE. Retinal pigment epithelial (RPE) cells have an important role in the immune suppression associated with the immune privilege of the eye. Some aspects of this remain unclear and this study aimed to determine how RPE cells could influence the production of chemokines by T lymphocytes.METHODS. T lymphocytes, separated from peripheral blood of normal volunteers, and RPE cells, cultured from donor eyes, were cultured separately and together, either in contact or in transwells. Supernatants were analyzed for CCL3, CCL4, and soluble CD54 (sCD54) by ELISA. Blocking agents were used to determine which soluble mediators were involved.RESULTS. Coculture of RPE cells with activated lymphocytes resulted in a reduction in CCL3 and CCL4 production by lymphocytes, primarily by soluble mediators. Soluble CD54 was markedly increased on coculture of lymphocytes with RPE cells. Soluble CD54 reduced CCL3 and CCL4 production by RPE cells, and inhibition of CCL3 and CCL4 on coculture with RPE cells was reduced by anti-CD54. Blocking prostaglandin E2 (PGE2) abrogated the inhibition of CCL4, but not CCL3, by RPE cells. Blocking TGF beta and nitric oxide production had no effect.CONCLUSIONS. RPE cells are able to down-regulate high levels of CCL3 and CCL4 production by T lymphocytes by using the soluble mediators sCD54 and PGE2. Reducing this production of CCL3 and CCL4 will dampen down the cascade effect and recruitment of more inflammatory cells, protecting the retina from an excessive immune response. (Invest Ophthalmol Vis Sci. 2013; 54: 722-730) DOI:10.1167/iovs.12-10602",

keywords = "intercellular-adhesion molecule-1, nitric-oxide synthase, experimental autoimmune enecephalomyelitis, function-associated antigen-1, gene-expression, inflammatory protein-1-alpha, endothelial-cells, immune privilege, beta-chemokines, soluble ICAM-1",

author = "Wallace, {Carol A.} and Gill Moir and Malone, {David F. G.} and Linda Duncan and Gayathri Devarajan and Crane, {Isabel J.}",

year = "2013",

month = jan,

day = "28",

doi = "10.1167/iovs.12-10602",

language = "English",

volume = "54",

pages = "722--730",

journal = "Investigative Ophthalmology & Visual Science",

issn = "0146-0404",

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

number = "1",

}

TY - JOUR

T1 - Regulation of T-lymphocyte CCL3 and CCL4 production by retinal pigment epithelial cells

AU - Wallace, Carol A.

AU - Moir, Gill

AU - Malone, David F. G.

AU - Duncan, Linda

AU - Devarajan, Gayathri

AU - Crane, Isabel J.

PY - 2013/1/28

Y1 - 2013/1/28

N2 - PURPOSE. Retinal pigment epithelial (RPE) cells have an important role in the immune suppression associated with the immune privilege of the eye. Some aspects of this remain unclear and this study aimed to determine how RPE cells could influence the production of chemokines by T lymphocytes.METHODS. T lymphocytes, separated from peripheral blood of normal volunteers, and RPE cells, cultured from donor eyes, were cultured separately and together, either in contact or in transwells. Supernatants were analyzed for CCL3, CCL4, and soluble CD54 (sCD54) by ELISA. Blocking agents were used to determine which soluble mediators were involved.RESULTS. Coculture of RPE cells with activated lymphocytes resulted in a reduction in CCL3 and CCL4 production by lymphocytes, primarily by soluble mediators. Soluble CD54 was markedly increased on coculture of lymphocytes with RPE cells. Soluble CD54 reduced CCL3 and CCL4 production by RPE cells, and inhibition of CCL3 and CCL4 on coculture with RPE cells was reduced by anti-CD54. Blocking prostaglandin E2 (PGE2) abrogated the inhibition of CCL4, but not CCL3, by RPE cells. Blocking TGF beta and nitric oxide production had no effect.CONCLUSIONS. RPE cells are able to down-regulate high levels of CCL3 and CCL4 production by T lymphocytes by using the soluble mediators sCD54 and PGE2. Reducing this production of CCL3 and CCL4 will dampen down the cascade effect and recruitment of more inflammatory cells, protecting the retina from an excessive immune response. (Invest Ophthalmol Vis Sci. 2013; 54: 722-730) DOI:10.1167/iovs.12-10602

AB - PURPOSE. Retinal pigment epithelial (RPE) cells have an important role in the immune suppression associated with the immune privilege of the eye. Some aspects of this remain unclear and this study aimed to determine how RPE cells could influence the production of chemokines by T lymphocytes.METHODS. T lymphocytes, separated from peripheral blood of normal volunteers, and RPE cells, cultured from donor eyes, were cultured separately and together, either in contact or in transwells. Supernatants were analyzed for CCL3, CCL4, and soluble CD54 (sCD54) by ELISA. Blocking agents were used to determine which soluble mediators were involved.RESULTS. Coculture of RPE cells with activated lymphocytes resulted in a reduction in CCL3 and CCL4 production by lymphocytes, primarily by soluble mediators. Soluble CD54 was markedly increased on coculture of lymphocytes with RPE cells. Soluble CD54 reduced CCL3 and CCL4 production by RPE cells, and inhibition of CCL3 and CCL4 on coculture with RPE cells was reduced by anti-CD54. Blocking prostaglandin E2 (PGE2) abrogated the inhibition of CCL4, but not CCL3, by RPE cells. Blocking TGF beta and nitric oxide production had no effect.CONCLUSIONS. RPE cells are able to down-regulate high levels of CCL3 and CCL4 production by T lymphocytes by using the soluble mediators sCD54 and PGE2. Reducing this production of CCL3 and CCL4 will dampen down the cascade effect and recruitment of more inflammatory cells, protecting the retina from an excessive immune response. (Invest Ophthalmol Vis Sci. 2013; 54: 722-730) DOI:10.1167/iovs.12-10602

KW - intercellular-adhesion molecule-1

KW - nitric-oxide synthase

KW - experimental autoimmune enecephalomyelitis

KW - function-associated antigen-1

KW - gene-expression

KW - inflammatory protein-1-alpha

KW - endothelial-cells

KW - immune privilege

KW - beta-chemokines

KW - soluble ICAM-1

U2 - 10.1167/iovs.12-10602

DO - 10.1167/iovs.12-10602

M3 - Article

SN - 0146-0404

VL - 54

SP - 722

EP - 730

JO - Investigative Ophthalmology & Visual Science

JF - Investigative Ophthalmology & Visual Science

IS - 1

ER -

Regulation of T-lymphocyte CCL3 and CCL4 production by retinal pigment epithelial cells

Abstract

Keywords

Access to Document

Fingerprint

Equipment

Iain Fraser Cytometry Centre

Cite this