May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Use of Chimeric Allografts to Elucidate the Role of Graft Epithelium versus Graft Stroma and Endothelium in Allosensitization.
Author Affiliations & Notes
  • D. R. Saban
    Ophthalmology, Schepens Eye Research Institute/Harvard Medical School, Boston, Massachusetts
  • S. K. Chauhan
    Ophthalmology, Schepens Eye Research Institute/Harvard Medical School, Boston, Massachusetts
  • J. El Annan
    Ophthalmology, Schepens Eye Research Institute/Harvard Medical School, Boston, Massachusetts
  • R. Dana
    Ophthalmology, Schepens Eye Research Institute/Harvard Medical School, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  D.R. Saban, None; S.K. Chauhan, None; J. El Annan, None; R. Dana, None.
  • Footnotes
    Support  5R01EY012963-08
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3719. doi:https://doi.org/
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      D. R. Saban, S. K. Chauhan, J. El Annan, R. Dana; Use of Chimeric Allografts to Elucidate the Role of Graft Epithelium versus Graft Stroma and Endothelium in Allosensitization.. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3719. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : All layers of a corneal allograft are not created equal. Graft epithelium is currently considered to bear the immunogenic load of a donor button, which is in part because it houses the majority of all corneal cells. However, this may not be exactly the case since epithelial rejection and/or replacement with host epithelial cells could diminish its immunogenicity. Thus, we hypothesized that graft stroma/endothelium also contributes significantly to alloimmunogenicity and is thereby important in allosensitization of the host.

Methods: : To address this, we relied on the orthotopic transplantation of chimeric corneal allografts, whereby C57BL/6 graft stroma/endothelium reconstituted with C3H/He graft epithelium (or vica versa) was placed onto a BALB/c host. Conventional C3H/He or C57BL/6 corneal allografts were placed onto other BALB/c hosts as well. Harvested T cells from host draining lymph nodes were co-cultured with BALB/c, C57BL/6, or C3H/He APC for 5 days. Allosensitization was subsequently measured by T cell proliferation via BrdU incorporation, IFN-γ secretion via ELISA, and CD4+ IFN-γ+ T cell frequencies via FACS analysis.

Results: : Conventional C57BL/6 allografts replaced with a C3H/He epithelium versus C3H/He stroma/endothelium resulted in higher T cell proliferation to C57BL/6 APC stimulation (increase of 40% versus 13% respectively, over the naïve control). Similarly, levels of culture supernatant IFN-γ to C57BL/6 APC stimulation increased by 22% versus 11% over the naïve control, respectively. Lastly, CD4+ IFN-γ+ T cell frequencies arising from C57BL/6 APC stimulation increased by 60% versus 17% over the naïve control, respectively. Similar data was also obtained throughout to C3H/He APC stimulation, when conventional C3H/He allografts were replaced with C57BL/6 epithelium versus C57BL/6 stroma/endothelium.

Keywords: cornea: basic science • transplantation • antigen presentation/processing 
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