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Effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection

  • Cornea
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Abstract

Background

Corneal graft rejection is the major cause of corneal failures. Previous studies have shown that the CD25 monoclonal antibody can inhibit corneal graft rejection during the acute phase of rejection in rat models. In the current study, we evaluated the safety and efficacy of the topical Entranster™ vector in rat corneal applications and further investigated the effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection.

Methods

Fluorescence detection, clinical assessment, hematoxylin and eosin (HE), terminal deoxynucleotidyl transferase biotin-dUTP nick end labelling (TUNEL) and CD11b assays were used to evaluate the safety and efficacy of CD25siRNA gene transfer in normal SD rat corneas. Orthotopic corneal transplants were performed in alkali burned SD rats. Corneal recipients were divided into four groups that received different treatments. Clinical assessment, western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry, HE and transmission electron microscopy (TEM) were performed on all grafts.

Results

Low toxicity, no immunogenicity and high transfection efficiency were observed in rat corneas treated with the Entranster™ vector. Reduced endothelial cell apoptosis, inflammatory cell infiltration and graft neovascularisation were observed in the CD25siRNA treatment groups. The graft survival curves showed that CD25siRNA treatment significantly prolonged graft survival time, with better graft transparency and less graft oedema. Lower CD25 and higher IL-10 expression were detected in the CD25siRNA treatment groups during the study period, and a higher FOXP3 level was found in the CD25siRNA group than in the CD25siRNA-twice group on days 14 and 21 post-operation.

Conclusions

The Entranster™ vector is an effective vector for corneal gene therapy. CD25 siRNA gene transfer inhibits corneal graft rejection via upregulation of anti-inflammatory molecule expression.

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Acknowledgments

This study was supported by funding from the National Natural Science Foundation of China (81170822).

Conflict of interest

All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

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Authors and Affiliations

  1. Department of Ophthalmology, First Teaching Hospital, Chongqing University of Medical Sciences, Chongqing Eye Bank, Chongqing, China

    Qin Qin, Yunjie Shi, Qingqing Zhao, Dan Luo, Yuan Chen, Jing Wu & Min Zhao

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  1. Qin Qin
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  2. Yunjie Shi
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  3. Qingqing Zhao
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  4. Dan Luo
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  5. Yuan Chen
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  6. Jing Wu
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  7. Min Zhao
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Correspondence to Min Zhao.

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Qin, Q., Shi, Y., Zhao, Q. et al. Effects of CD25siRNA gene transfer on high-risk rat corneal graft rejection. Graefes Arch Clin Exp Ophthalmol 253, 1765–1776 (2015). https://doi.org/10.1007/s00417-015-3067-2

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  • DOI: https://doi.org/10.1007/s00417-015-3067-2

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