Leukocyte trafficking in experimental autoimmune uveitis: breakdown of blood-retinal barrier and upregulation of cellular adhesion molecules

Heping Xu, John Vincent Forrester, Janet Mary Liversidge, Isabel Joan Crane

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

PURPOSE. To clarify the order of events occurring in the breakdown of the blood-retinal barrier (BRB) in experimental autoimmune uveoretinitis (EAU) and in particular to study the relationships between increased vascular permeability, upregulation of endothelial cell adhesion molecules, and leukocyte adhesion and infiltration during EAU.

METHODs. B10.RIII mice were immunized with human interphotoreceptor retinoid binding protein (IRBP) peptide 161-180. Changes in the retinal microvasculature were examined on days 3, 6, 7, 8, 9, 10, 16, and 21 postimmunization (pi). Evans blue dye was administered intravenously to assess vascular permeability. Expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, P-selectin, E-selectin, and platelet endothelial cell adhesion molecule (PECAM)-1 was evaluated by in vivo administration of antibody and subsequent immunostaining of retinal whole-mounts. Lymphocytes from inguinal lymph nodes of normal and chicken ovalbumin (OVA)- or IRBP peptide-immunized mice at day 5, 6, 7, 8, and 15 pi were labeled in vitro with calcein-AM (C-AM) and infused intravenously into syngeneic recipient mice, which had been immunized with peptide at the same corresponding time point. Wholemount preparations of retinas were observed 24 hours later by confocal microscopy to determine the adhesion and infiltration of lymphocytes.

RESULTS. The first observation of an increase in vascular permeability occurred at day 7 pi and was restricted to focal areas of the retinal postcapillary venules of the inner vascular plexus. This progressively extended to the outer vascular plexus at day 9 pi. Specific adhesion of leukocytes to the endothelium of retinal venules of the inner vascular plexus was first observed at day 6 pi. Leukocyte extravasation into the retinal parenchyma from these vessels began at day 8 pi and extended to the outer vascular plexus at day 9 pi. The expression of adhesion molecules increased progressively during the development of EAU. In particular, the adhesion molecules ICAM-1, P-selectin, and E-selectin were expressed predominately in retinal venules, the sites of BRB breakdown, cell adhesion, and extravasation, from day 7 pi. The increases in expression of ICAM-1 and P-selectin were associated both spatially and temporally with breakdown of the BRB, cell adhesion, and extravasation. No increase in expression of P-selectin and ICAM-1 was observed in either the mesenteric vessels of EAU mice or the retinal vessels of OVA-immunized mice.

CONCLUSIONS. The sequence of events in EAU appears to be focal adhesion of leukocytes to discrete sites on postcapillary venules, followed by upregulation of adhesion molecules, especially ICAM-1 and P-selectin, and breakdown of the BRB, leading to transendothelial migration of leukocytes and recruitment of large numbers of cells to the retinal parenchyma. These changes occur over a short period of 6 to 9 days pi and initiate the process of tissue damage during the following 2 to 3 weeks.

Original languageEnglish
Pages (from-to)226-234
Number of pages8
JournalInvestigative Ophthalmology & Visual Science
Volume44
Issue number1
DOIs
Publication statusPublished - Jan 2003

Keywords

  • NECROSIS-FACTOR-ALPHA
  • MONOCLONAL-ANTIBODIES
  • PECAM-1 CD31
  • S-ANTIGEN
  • IN-VIVO
  • UVEORETINITIS
  • MIGRATION
  • LFA-1
  • PERMEABILITY
  • INFILTRATION

Cite this

Leukocyte trafficking in experimental autoimmune uveitis: breakdown of blood-retinal barrier and upregulation of cellular adhesion molecules. / Xu, Heping; Forrester, John Vincent; Liversidge, Janet Mary; Crane, Isabel Joan.

In: Investigative Ophthalmology & Visual Science, Vol. 44, No. 1, 01.2003, p. 226-234.

Research output: Contribution to journalArticle

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author = "Heping Xu and Forrester, {John Vincent} and Liversidge, {Janet Mary} and Crane, {Isabel Joan}",
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TY - JOUR

T1 - Leukocyte trafficking in experimental autoimmune uveitis: breakdown of blood-retinal barrier and upregulation of cellular adhesion molecules

AU - Xu, Heping

AU - Forrester, John Vincent

AU - Liversidge, Janet Mary

AU - Crane, Isabel Joan

PY - 2003/1

Y1 - 2003/1

N2 - PURPOSE. To clarify the order of events occurring in the breakdown of the blood-retinal barrier (BRB) in experimental autoimmune uveoretinitis (EAU) and in particular to study the relationships between increased vascular permeability, upregulation of endothelial cell adhesion molecules, and leukocyte adhesion and infiltration during EAU.METHODs. B10.RIII mice were immunized with human interphotoreceptor retinoid binding protein (IRBP) peptide 161-180. Changes in the retinal microvasculature were examined on days 3, 6, 7, 8, 9, 10, 16, and 21 postimmunization (pi). Evans blue dye was administered intravenously to assess vascular permeability. Expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, P-selectin, E-selectin, and platelet endothelial cell adhesion molecule (PECAM)-1 was evaluated by in vivo administration of antibody and subsequent immunostaining of retinal whole-mounts. Lymphocytes from inguinal lymph nodes of normal and chicken ovalbumin (OVA)- or IRBP peptide-immunized mice at day 5, 6, 7, 8, and 15 pi were labeled in vitro with calcein-AM (C-AM) and infused intravenously into syngeneic recipient mice, which had been immunized with peptide at the same corresponding time point. Wholemount preparations of retinas were observed 24 hours later by confocal microscopy to determine the adhesion and infiltration of lymphocytes.RESULTS. The first observation of an increase in vascular permeability occurred at day 7 pi and was restricted to focal areas of the retinal postcapillary venules of the inner vascular plexus. This progressively extended to the outer vascular plexus at day 9 pi. Specific adhesion of leukocytes to the endothelium of retinal venules of the inner vascular plexus was first observed at day 6 pi. Leukocyte extravasation into the retinal parenchyma from these vessels began at day 8 pi and extended to the outer vascular plexus at day 9 pi. The expression of adhesion molecules increased progressively during the development of EAU. In particular, the adhesion molecules ICAM-1, P-selectin, and E-selectin were expressed predominately in retinal venules, the sites of BRB breakdown, cell adhesion, and extravasation, from day 7 pi. The increases in expression of ICAM-1 and P-selectin were associated both spatially and temporally with breakdown of the BRB, cell adhesion, and extravasation. No increase in expression of P-selectin and ICAM-1 was observed in either the mesenteric vessels of EAU mice or the retinal vessels of OVA-immunized mice.CONCLUSIONS. The sequence of events in EAU appears to be focal adhesion of leukocytes to discrete sites on postcapillary venules, followed by upregulation of adhesion molecules, especially ICAM-1 and P-selectin, and breakdown of the BRB, leading to transendothelial migration of leukocytes and recruitment of large numbers of cells to the retinal parenchyma. These changes occur over a short period of 6 to 9 days pi and initiate the process of tissue damage during the following 2 to 3 weeks.

AB - PURPOSE. To clarify the order of events occurring in the breakdown of the blood-retinal barrier (BRB) in experimental autoimmune uveoretinitis (EAU) and in particular to study the relationships between increased vascular permeability, upregulation of endothelial cell adhesion molecules, and leukocyte adhesion and infiltration during EAU.METHODs. B10.RIII mice were immunized with human interphotoreceptor retinoid binding protein (IRBP) peptide 161-180. Changes in the retinal microvasculature were examined on days 3, 6, 7, 8, 9, 10, 16, and 21 postimmunization (pi). Evans blue dye was administered intravenously to assess vascular permeability. Expression of intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, P-selectin, E-selectin, and platelet endothelial cell adhesion molecule (PECAM)-1 was evaluated by in vivo administration of antibody and subsequent immunostaining of retinal whole-mounts. Lymphocytes from inguinal lymph nodes of normal and chicken ovalbumin (OVA)- or IRBP peptide-immunized mice at day 5, 6, 7, 8, and 15 pi were labeled in vitro with calcein-AM (C-AM) and infused intravenously into syngeneic recipient mice, which had been immunized with peptide at the same corresponding time point. Wholemount preparations of retinas were observed 24 hours later by confocal microscopy to determine the adhesion and infiltration of lymphocytes.RESULTS. The first observation of an increase in vascular permeability occurred at day 7 pi and was restricted to focal areas of the retinal postcapillary venules of the inner vascular plexus. This progressively extended to the outer vascular plexus at day 9 pi. Specific adhesion of leukocytes to the endothelium of retinal venules of the inner vascular plexus was first observed at day 6 pi. Leukocyte extravasation into the retinal parenchyma from these vessels began at day 8 pi and extended to the outer vascular plexus at day 9 pi. The expression of adhesion molecules increased progressively during the development of EAU. In particular, the adhesion molecules ICAM-1, P-selectin, and E-selectin were expressed predominately in retinal venules, the sites of BRB breakdown, cell adhesion, and extravasation, from day 7 pi. The increases in expression of ICAM-1 and P-selectin were associated both spatially and temporally with breakdown of the BRB, cell adhesion, and extravasation. No increase in expression of P-selectin and ICAM-1 was observed in either the mesenteric vessels of EAU mice or the retinal vessels of OVA-immunized mice.CONCLUSIONS. The sequence of events in EAU appears to be focal adhesion of leukocytes to discrete sites on postcapillary venules, followed by upregulation of adhesion molecules, especially ICAM-1 and P-selectin, and breakdown of the BRB, leading to transendothelial migration of leukocytes and recruitment of large numbers of cells to the retinal parenchyma. These changes occur over a short period of 6 to 9 days pi and initiate the process of tissue damage during the following 2 to 3 weeks.

KW - NECROSIS-FACTOR-ALPHA

KW - MONOCLONAL-ANTIBODIES

KW - PECAM-1 CD31

KW - S-ANTIGEN

KW - IN-VIVO

KW - UVEORETINITIS

KW - MIGRATION

KW - LFA-1

KW - PERMEABILITY

KW - INFILTRATION

U2 - 10.1167/iovs.01-1202

DO - 10.1167/iovs.01-1202

M3 - Article

VL - 44

SP - 226

EP - 234

JO - Investigative Ophthalmology & Visual Science

JF - Investigative Ophthalmology & Visual Science

SN - 0146-0404

IS - 1

ER -