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T lymphocyte mediated lysis of mitomycin C treated Tenon’s capsule fibroblasts

¹ Wound Healing Research Unit, Institute of Ophthalmology, Bath Street, London EC1V 9EL, UK
² Glaucoma Unit, Moorfields Eye Hospital, City Road, London EC1V 2PD, UK
³ Department of Clinical Immunology, Royal Free Hospital School of Medicine, Pond Street, London NW3, UK

Correspondence to:
Correspondence to:
J G Crowston
Hamilton Glaucoma Center, UCSD, 9500 Gilman Drive, La Jolla, CA 92093-0946, USA; jcrowston{at}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.

Keywords: apoptosis; glaucoma; T cells; Tenon’s fibroblasts; wound healing

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