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Updated: Aug 26, 2026

Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes
Published on: June 16, 2011
T lymphocyte mediated lysis of mitomycin C treated Tenon's capsule fibroblasts
J G Crowston1, L H Chang, J T Daniels
1Wound Healing Research Unit, Institute of Ophthalmology, Bath Street, London EC1V 9EL, UK. jcrowston@ucsd.edu
Aims:
To evaluate the effect of T cell co-culture on mitomycin C treated and untreated Tenon's capsule fibroblasts.
Methods:
IL-2 dependent allogeneic T cells were incubated over a monolayer of mitomycin C treated or control fibroblasts. Fibroblast numbers were evaluated by direct counts using phase contrast microscopy. To determine whether T cell mediated lysis was a consequence of MHC mismatch, co-culture experiments were repeated with autologous T cells. The effect of Fas receptor blockade was established by co-incubation with a Fas blocking (M3) antibody.
Results:
T cell co-culture resulted in a dramatic reduction in fibroblast survival compared to mitomycin C treatment alone (p = 0.032). T cell killing required fibroblast/lymphocyte cell to cell contact and was observed in both allogeneic and autologous co-culture experiments. Fas blocking antibodies did not significantly inhibit T cell killing (p = 0.39).
Conclusion:
T cells augment mitomycin C treated fibroblast death in vitro. Similar mechanisms may contribute to the cytotoxic effect of mitomycin C in vivo and account for the largely hypocellular drainage blebs that are observed clinically.
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