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Influence of combined treatment of low dose rapamycin and cyclosporin A on corneal allograft survival

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Abstract

Purpose

To analyze the immune modulatory effect of low-dose systemic treatment with rapamycin (Rapa) alone or in combination with cyclosporin A (CsA) in a high-responder corneal allograft model.

Methods

A total of 80 C57BL/6 mice received corneal grafts from BALB/c donors. Recipients were treated with either CsA 3 mg/kg/day or Rapa 0.5 mg/kg/day monotherapy or received combined treatment. Immunomodulatory treatment was started on the day of surgery, and continued for 14 days. The frequency of CD4+CD25+Foxp3+ T regulatory cells (Treg) in secondary lymphoid organs was measured by flow cytometry. Development of IFN-γ producing alloreactive T cells was estimated by Elispot. In addition, corneal samples were subjected to real-time RT-PCR analysis for cytokine transcription.

Results

Monotherapy with Rapa significantly delayed allograft rejection (13.4 ± 1.34 days, p = 0.03). However, the combination of both, low-dose Rapa and CsA prolonged corneal allograft survival at a significantly higher level (MST = 17.1 ± 1.37 days, p = 0.0001) than in the control group (MST = 11.2 ± 1.91 days). Rapa monotherapy increased the frequency of CD4+CD25+Foxp3+Treg in draining lymph nodes, whereas addition of CsA reduced Tregs. Monotherapy with Rapa as well as combined treatment prevented development of IFN-γ producing alloreactive T cells in spleen. Combined treatment resulted in down-regulation of intragraft CD3, IL-2, IFN-γ and IL-10 transcription (p = 0.028, p = 0.027, p = 0.028 and p = 0.027 respectively).

Conclusions

Combined treatment with low-dose CsA and Rapa resulted in superior graft survival, and effectively modulated mRNA expression of inflammation and infiltration markers.

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Authors and Affiliations

  1. Department of Ophthalmology, Charité University Medicine, 10117, Berlin, Germany

    Svetlana Stanojlovic & Uwe Pleyer

  2. Institute of Medical Immunology, Charité University Medicine, Berlin, Germany

    Stephan Schlickeiser, Christine Appelt, Katrin Vogt, Isabela Schmitt-Knosalla, Stefanie Haase & Birgit Sawitzki

  3. Regenerative Medicine Institute (REMEDI), National Centre for Biomedical Engineering Science (NCBES), National University of Ireland, Galway, Ireland

    Thomas Ritter

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  1. Svetlana Stanojlovic
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  2. Stephan Schlickeiser
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  3. Christine Appelt
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  5. Isabela Schmitt-Knosalla
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Correspondence to Uwe Pleyer.

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This work was supported by Deutsche Forschungsgemeinschaft (Pl 150/14-2) and Sonderforschungsbereich (SFB650 TP14).

Birgit Sawitzki and Uwe Pleyer made an equal contribution

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Stanojlovic, S., Schlickeiser, S., Appelt, C. et al. Influence of combined treatment of low dose rapamycin and cyclosporin A on corneal allograft survival. Graefes Arch Clin Exp Ophthalmol 248, 1447–1456 (2010). https://doi.org/10.1007/s00417-010-1420-z

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  • DOI: https://doi.org/10.1007/s00417-010-1420-z

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