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

Am J Vet Res. 2002 Jul;63(7):942-7. doi: 10.2460/ajvr.2002.63.942.

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-1beta and tumor necrosis factor-alpha.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aqueous Humor / metabolism
  • Chemokine CCL5 / biosynthesis*
  • Chemokine CCL5 / genetics
  • Chemotaxis
  • Ciliary Body / cytology
  • Ciliary Body / metabolism*
  • Epithelial Cells / metabolism
  • Female
  • Horse Diseases / metabolism*
  • Horses
  • Male
  • RNA / chemistry
  • RNA / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sequence Analysis, DNA
  • Uveitis / metabolism
  • Uveitis / veterinary*

Substances

  • Chemokine CCL5
  • RNA