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Expression of a chemokine by ciliary body epithelium in horses with naturally occurring recurrent uveitis and in cultured ciliary body epithelial cells

Brian C. GilgerComparative Ophthalmology Research Laboratories, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606.

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 DVM, MS
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Ping YangDuke University Eye Center, Department of Ophthalmology, College of Medicine, Duke University, Durham, NC 27705.

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 MD, PhD
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Jacklyn H. SalmonComparative Ophthalmology Research Laboratories, Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606.

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 BS
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Glenn J. JaffeDuke University Eye Center, Department of Ophthalmology, College of Medicine, Duke University, Durham, NC 27705

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Janice B. AllenDepartment of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606.

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 PhD

Abstract

Objective—To determine whether a chemokine (RANTES)-like protein expressed by ciliary epithelium plays a role in uveitis.

Sample Population—3 clinically normal horses intradermal, 5 eyes from 5 horses with recurrent uveitis, and 10 normal eyes from 5 age- and sex-matched horses.

Procedure—Cross-reactivity and sensitivity of recombinant human (rh)-regulated upon activation, normal T-cell expressed and secreted (RANTES) protein were evaluated in horses by use of intradermal hypersensitivity reactions and a chemotaxis assay. Aqueous humor and ciliary body of eyes from clinically normal horses and horses with uveitis were examined for RANTES expression by use of an ELISA and reverse transcription-polymerase chain reaction (RT-PCR). Expression of RANTES mRNA and protein content of primary cultures of equine ciliary pigmented epithelial cells (RT-PCR) and culture supernatant (ELISA) were measured 6 or 24 hours, respectively, after cultures were stimulated with interleukin-1β and tumor necrosis factor-α.

Results—Strong reactions to intradermal hypersensitivity testing and significant chemotaxis of equine leukocytes to rh-RANTES wereas observed. Aqueous humor of eyes from horses with uveitis contained increased concentrations of rh-RANTES-like protein (mean ± SD, 45.9 ± 31.7 pg/ml), compared with aqueous humor from clinically normal horses (0 pg/ml). Ciliary body from horses with uveitis expressed RANTES mRNA, whereas ciliary body from clinically normal horses had low mRNA expression. Stimulated ciliary pigmented epithelial cells expressed increased amounts of rh-RANTES-like protein (506.1 ± 298.3 pg/ml) and mRNA, compared with unstimulated samples.

Conclusions and Clinical Relevance—Ciliary epithelium may play a role in recruitment and activation of leukocytes through expression of RANTES. (Am J Vet Res 2002;63:942–947)

Abstract

Objective—To determine whether a chemokine (RANTES)-like protein expressed by ciliary epithelium plays a role in uveitis.

Sample Population—3 clinically normal horses intradermal, 5 eyes from 5 horses with recurrent uveitis, and 10 normal eyes from 5 age- and sex-matched horses.

Procedure—Cross-reactivity and sensitivity of recombinant human (rh)-regulated upon activation, normal T-cell expressed and secreted (RANTES) protein were evaluated in horses by use of intradermal hypersensitivity reactions and a chemotaxis assay. Aqueous humor and ciliary body of eyes from clinically normal horses and horses with uveitis were examined for RANTES expression by use of an ELISA and reverse transcription-polymerase chain reaction (RT-PCR). Expression of RANTES mRNA and protein content of primary cultures of equine ciliary pigmented epithelial cells (RT-PCR) and culture supernatant (ELISA) were measured 6 or 24 hours, respectively, after cultures were stimulated with interleukin-1β and tumor necrosis factor-α.

Results—Strong reactions to intradermal hypersensitivity testing and significant chemotaxis of equine leukocytes to rh-RANTES wereas observed. Aqueous humor of eyes from horses with uveitis contained increased concentrations of rh-RANTES-like protein (mean ± SD, 45.9 ± 31.7 pg/ml), compared with aqueous humor from clinically normal horses (0 pg/ml). Ciliary body from horses with uveitis expressed RANTES mRNA, whereas ciliary body from clinically normal horses had low mRNA expression. Stimulated ciliary pigmented epithelial cells expressed increased amounts of rh-RANTES-like protein (506.1 ± 298.3 pg/ml) and mRNA, compared with unstimulated samples.

Conclusions and Clinical Relevance—Ciliary epithelium may play a role in recruitment and activation of leukocytes through expression of RANTES. (Am J Vet Res 2002;63:942–947)