T-bet expression in the iris and spleen parallels disease expression during endotoxin-induced uveitis

B. Li, P. Yang, L. Chu, H. Zhou, X. Huang, L. Zhu, A. Kijlstra

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

T lymphocytes have been implicated in the development of endotoxin-induced uveitis (EIU). T-bet is a Th1 cell-specific transcription factor that is involved in differentiation and effector functions. The aim of this study was to investigate kinetics of T-bet expression at the mRNA and protein levels during EIU using real-time PCR and whole-mount immunohistochemistry. Methods A single footpad injection of 200 ¿g of lipopolysaccharide (LPS) was administered to male Wistar rats in order to induce EIU. Clinical changes were followed by slit-lamp examination. The expression of T-bet mRNA in the spleen was evaluated 0, 8, 16, 24, 48, and 96 h after LPS injection using real-time PCR. Immunohistochemistry was performed on the iris whole-mounts as well as on frozen sections of the spleen to evaluate T-bet protein expression. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was performed on the iris whole-mounts to assay apoptotic cells. Results Uveitis was observed in all rats that received LPS. T-bet+ cells and TUNEL+ cells in the iris whole-mounts showed a similar pattern in cell number and distribution and both types of cells were observed at 8 h, significantly increased 24 h, and decreased 48 h after LPS injection. T-bet expression at both the mRNA and protein levels in spleen also paralleled ocular inflammation. It was weakly detectable after 0 h, increased after 8 h (index 1.3, T-bet+ cells OD 17.43±2.15), reached its peak after 24 h (index 4.00, OD 53.52±4.00), and decreased 48 h following LPS injection (index 1.38, OD 25.75±2.45). Conclusions The results show that T-bet expression in both the iris and the spleen, and in apoptotic cells in the iris parallel the severity of intraocular inflammation after systemic LPS administration. These results suggest that T-bet may play a significant role in the dynamics of EIU.
Original languageEnglish
Pages (from-to)407-413
JournalGraefes Archive for Clinical and Experimental Ophthalmology
Volume245
Issue number3
DOIs
Publication statusPublished - 2007

Keywords

  • transcription factor
  • ocular inflammation
  • lineage commitment
  • interferon-gamma
  • dendritic cells
  • ciliary body
  • rat
  • macrophages
  • injection
  • cytokines

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