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Samantha Herretes, Juan Murillo, Duncan Ross, Henry Barreras, Yaohong Tan, Ali Saeed, Astrid Gonzalez, Carolina Betancurt, Robert Levy, Victor Perez; Demonstration of Donor T cell Recruitment and Correlation with Inflammatory Cytokine Presence in Experimental Ocular Graft-versus-Host Disease. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4520.
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Graft-versus-host disease is a potentially blinding condition that may affect allogeneic hematopoietic stem cell transplant (HSCT) survivors. Alloreactive donor T cells are essential for the development of GVHD, but their direct role in local ocular GVHD has not been demonstrated. In this study, we examined immunological and ocular changes using a clinically relevant MHC-matched, minor antigen mis-matched HSCT transplantation model of systemic and ocular GVHD.
After high-dose TBI, C3H.SW (H2b) mice were transplanted with TCD-BM alone or together with T cells from either wild type B6 mice or CXCR6-GFP knockin mice, which express GFP regulated by the CXCR6 promoter (B6-CXCR6-/- GFP). GVHD was monitored weekly by clinical score composite and ocular surface fluorescein staining. In vivo fluorescent microscopy was used to measure the recruitment of B6-CXCR6-/- GFP donor T cells. At 6-7 weeks goblet cells were quantified using PAS staining and T cell phenotype was characterized by flow cytometry analysis of corneal cells suspension. Total RNA was prepared and the production of mediators of inflammation analyzed by qPCR.
Mice that received transplanted BM and T-cells from either wild type B6 or B6-CXCR6-/- GFP developed evidence of systemic GVHD by day 21 as opposed to control animals. Beginning at day 28, the recruitment of donor B6-CXCR6-/- GFP T cells correlated with the development of progressively increasing ocular surface disease in mice with GVHD. Interesting, relative expression of RNA of the T cell chemo-attractant CXCL10 and its receptor CXCR3, also increased in the corneas. Similarly, the relative expression of the inflammatory cytokines, IFN-γ TNF- α and IL-6 were significantly increased, correlating with a significantly decreased number of goblet cells in the conjunctiva of mice with GVHD but not controls.
C3H.SW mice that receive BM with T cells develop ocular GVHD characterized by donor T cell recruitment, increased expression of CXCL10/CXCR3, inflammatory cytokines and the development of corneal staining, ulceration and loss of goblet cells. These findings support a critical role for donor T cells in the immune responses occurring at the ocular surface. Understanding kinetics and pathways will provide targets for the prevention and treatment of ocular GVHD.
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