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Updated: Dec 9, 2025
Murine Full-thickness Skin Transplantation
Published on: January 2, 2017
DNA-PKcs Inhibition Extends Allogeneic Skin Graft Survival
David K Harrison1,2, Zachary J Waldrip1,2, Lyle Burdine1,3
1Division of Surgical Research, University of Arkansas for Medical Sciences, Little Rock, AR.
Background:
Organ transplantation is life-saving and continued investigations into immunologic mechanisms that drive organ rejection are needed to improve immunosuppression therapies and prevent graft failure. DNA-dependent protein kinase catalytic subunit, DNA dependent-protein kinase catalytic subunit (DNA-PKcs), is a critical component of both the cellular and humoral immune responses. In this study, we investigate the contribution of DNA-PKcs to allogeneic skin graft rejection to potentially highlight a novel strategy for inhibiting transplant rejection.
Methods:
Fully MHC mismatched murine allogeneic skin graft studies were performed by transplanting skin from BalbC mice to C57bl6 mice and treating with either vehicle or the DNA-PKcs inhibitor NU7441. Graft rejection, cytokine production, immune cell infiltration, and donor-specific antibody formation were analyzed.
Results:
DNA-PKcs inhibition significantly reduced necrosis and extended graft survival compared with controls (mean survival 14 d versus 9 d, respectively). Inhibition reduced the production of the cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, TNF-α, and IFN-γ and the infiltration of CD3+ lymphocytes into grafts. Furthermore, DNA-PKcs inhibition reduced the number of CD19+ B cells and CD19+ CD138+ plasma cells coinciding with a significant reduction in donor-specific antibodies. At a molecular level, we determined that the immunosuppressive effects of DNA-PKcs inhibition were mediated, in part, via inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells signaling through reduced expression of the p65 subunit.
Conclusions:
Our data confirm that DNA-PKcs contributes to allogeneic graft rejection and highlight a novel immunologic function for DNA-PKcs in the regulation of nuclear factor kappa-light-chain-enhancer of activated B cells and concomitant cytokine production.
Insights
Inhibiting DNA-PKcs, a key immune response protein, significantly reduced skin graft rejection in mice. This finding offers a potential new strategy for improving transplant success by modulating immune responses.
Area of Science:
- Immunology
- Transplantation Biology
- Molecular Medicine
Background:
- Organ transplantation is vital but challenged by immune-mediated graft rejection.
- DNA-dependent protein kinase catalytic subunit (DNA-PKcs) plays a role in cellular and humoral immunity.
- Understanding DNA-PKcs's role in rejection is crucial for developing better immunosuppression therapies.
Purpose of the Study:
- To investigate the role of DNA-PKcs in allogeneic skin graft rejection.
- To evaluate the potential of DNA-PKcs inhibition as a strategy to prevent transplant rejection.
Main Methods:
- Murine allogeneic skin grafts (BalbC to C57bl6) were used.
- Mice were treated with either a DNA-PKcs inhibitor (NU7441) or a vehicle control.
- Analysis included graft rejection, cytokine profiles, immune cell infiltration, and antibody production.
Main Results:
- DNA-PKcs inhibition significantly prolonged skin graft survival (14 days vs. 9 days).
- Inhibition reduced key pro-inflammatory cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ) and immune cell infiltration (CD3+ lymphocytes).
- Reduced B cell and plasma cell populations correlated with decreased donor-specific antibodies, mediated partly by inhibiting NF-κB signaling.
Conclusions:
- DNA-PKcs inhibition effectively reduces allogeneic skin graft rejection.
- DNA-PKcs has a novel immunoregulatory role in controlling cytokine production and NF-κB signaling in transplant rejection.
