The Charles T. Campbell Eye Microbiology Lab
UPMCUniversity of Pittsburgh Schools of the Health Sciences
HomeContact InformationLab Diagnostic TestingAntibiotic SusceptibilityAntimicrobial TherapyCurrent ResearchPhotos

Ocular Microbiology and Immunology Group
Back to OMIG Main Page

2014 Agenda and Abstracts | < Previous | Next >

2014 OMIG Abstract 19

The Contribution of T cells to Corneal Angiogenic Privilege in Transplantation
Antonio Di Zazzo, Brinda Subbarayal, Thomas Dohlman, Takenori Inomata, Sunil Chauhan, Reza Dana
Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary,
Department of Ophthalmology, Harvard Medical School, Boston, MA

Purpose: To investigate the effect of T effector cells (Teffs) and/or regulatory T cells (Tregs) from high-risk (HR) or low-risk (LR) transplanted mice on vascular endothelial cell (VEC) proliferation.

Methods: Ipsilateral lymph nodes were collected from Balb/c mice 14 days after low-risk and high-risk corneal transplantation. CD4+CD25-effector T cells and CD4+CD25+ Tregs, sorted by magnetic beads and stimulated with anti-CD3, were cocultured in a 96-well plate with MILE SVEN 1 cells, a vascular endothelial cell line, in DMEM/FBS10%. VEC proliferation was measured using the BrdU incorporation assay 19 hours after coculturing.

Results: CD4+CD25- Teffs from high-risk and low-risk transplanted mice significantly promoted VEC proliferation compared to positive (VEGF-A 20ng/ml) and negative controls. VEC proliferation was significantly higher when cocultured with Teffs from HR than Teffs from LR and naïve mice (Teff HR 0.9 ± 0.03;Teff LR 0.7234 ± 0.1; VEGF-A 0.6 ± 0.03; Negative Control: 0.5± 0.02 p<0.0001). After culturing Teffs and VECs with CD4+CD25+ Tregs from transplanted mice VEC proliferation was significantly decreased compared to VECs cultured with Teffs alone (Teff 0.9 ± 0.03; Teff+Treg 0.6 ± 0.06; p<0.0005). Tregs alone had no effect on VEC proliferation.

Conclusions: Our data showed a direct induction of VEC proliferation mediated by CD4+CD25- effector T cells isolated from high-risk and low risk transplanted mice. Further, Tregs indirectly inhibited Teff cell-mediated VEC proliferation. Our results suggest that a balance between pro-aniogenic Teffs and anti-angiogeneic Tregs is essential to sustain angiogenic privilege in HR corneal transplantation.

Disclosure: N
Grant Support: NIH EY 012963

2014 Agenda and Abstracts | < Previous | Next >