Passenger leucocytes and inflammatory debris transferred from the donor limb to the recipient can induce allorecognition, which activates the host immune response. This is the first study to evaluate whether the transfer of this inflammatory burden can be reduced via post-preservation flush prior to revascularisation, and whether this is influenced by ischaemia.
Bilateral forelimbs from the same pig were procured and infused with preservation flush and stored on ice. Each limb from the same pig underwent a post-preservation intravascular flush with isotonic solution at either 2 or 6 h. Venous effluent underwent flow cytometry to phenotype leucocyte populations, with additional quantification of cytokines and cell-free DNA.
We identified large populations of viable leucocytes in the flush effluent (8.65 × 108 ± 3.10 × 108 cells at 2 h and 1.02 × 109 ± 2.63 × 108 at 6 h). This comprised T cells, B cells, NK cells and monocytes. Post-preservation flush yielded significant concentrations of pro-inflammatory cytokines including IL-6, IL-18, GM-CSF, IL-1β, IL1α and CXCL-8 and mitochondrial DNA. The regulatory cytokine, IL-10 was undetectable.
This study supports the finding that a post-preservation flush removes leucocytes and inflammatory components that are responsible for direct presentation. This study also gives an indication of how ischaemia impacts on the inflammatory burden transferred to the recipient upon reperfusion.
To read this article in full you will need to make a payment
Purchase one-time access:Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
One-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:Subscribe to Journal of Plastic, Reconstructive & Aesthetic Surgery
Already a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
- Principalization of plastic surgery.1986
- Science of composite tissue allotransplantation.Transplantation. 2008; 86: 627-635
- Hand transplantation versus hand prosthetics: pros and cons.Curr Surg Rep. 2016; 4: 8
- Immunopathology of rejection: do the rules of solid organ apply to vascularized composite allotransplantation?.Curr Opin Organ Transplant. 2015; 20: 596-601
- The International Registry on Hand and Composite Tissue allotransplantation.Clin Transpl. 2011; : 247-253
- Direct versus indirect allorecognition pathways: on the right track.Am J Transplant. 2009; 9: 655-656
- Immunotherapy in transplantation: principles and practice.Wiley-Blackwell, 2012
- Altered immunogenicity of donor lungs via removal of passenger leukocytes using ex vivo lung perfusion.Am J Transplant. 2016; 16: 33-43
- Restoration of immunogenicity to passenger cell-depleted kidney allografts by the addition of donor strain dendritic cells.J Exp Med. 1982; 155: 31-41
- Human hand allograft: report on first 6 months.Lancet. 1999; 353: 1315-1320
- First Polish forearm transplantation: report after 17 months.Transplant Proc. 2009; 41: 549-553
- First forearm transplantation: outcome at 3 years.Am J Transplant. 2007; 7: 1753-1762
- The first (double) hand transplantation in Turkey.Transplant Proc. 2011; 43: 3557-3560
- Technical implications in proximal forearm transplantation.Tech Hand Up Extrem Surg. 2013; 17: 228-231
- Prolonged ischemic time, delayed graft function, and graft and patient outcomes in live donor kidney transplant recipients.Am J Transplant. 2016;
- Establishment of a swine model for validation of perfusion measurement by dynamic contrast-enhanced magnetic resonance imaging.Biomed Res Int. 2014; 2014: 390506
- The immunology of the porcine skin and its value as a model for human skin.Mol Immunol. 2015; 66: 14-21
- Evaluation of a porcine whole-limb heterotopic autotransplantation model.Microsurgery. 2013; 33: 141-147
- Characterizing the early inflammatory contribution of the donor kidney following reperfusion.Nephrol Dial Transplant. 2017;
- Haematology and biochemistry reference values for sows kept under modern management conditions.Vet Q. 1994; 16: 127-130
- Augmentation of recipient adaptive alloimmunity by donor passenger lymphocytes within the transplant.Cell Rep. 2016; 15: 1214-1227
- NK cells contribute to the skin graft rejection promoted by CD4+ T cells activated through the indirect allorecognition pathway.Int Immunol. 2008; 20: 1343-1349
- Immune recognition and rejection of allogeneic skin grafts.Immunotherapy. 2011; 3: 757-770
- NK cells promote transplant tolerance by killing donor antigen-presenting cells.J Exp Med. 2006; 203: 1851-1858
- Activation of transplant immunity: effect of donor leukocytes on thyroid allograft rejection.Science. 1976; 191: 385-388
- Prolonged cold ischemic time results in increased acute rejection in a rat allotransplantation model.J Surg Res. 2010; 164: e299-e304
- Ischemia-reperfusion injury in vascularized composite allotransplantation.J Craniofac Surg. 2013; 24: 51-56
- The inflammasome NLRs in immunity, inflammation, and associated diseases.Annu Rev Immunol. 2011; 29: 707-735
- Cytokine-activated human vascular smooth muscle delays apoptosis of neutrophils: relevance of interactions between cyclo-oxygenase-2 and colony-stimulating factors.FASEB J. 2001; 15: 1813-1815
Published online: November 24, 2017
Accepted: November 7, 2017
Received: April 19, 2017
Authored by a member of BAPRAS
© 2017 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.
ScienceDirectAccess this article on ScienceDirect
- Comment on “Ex-vivo flush of the limb allograft reduces inflammatory burden prior to transplantation”Journal of Plastic, Reconstructive & Aesthetic SurgeryVol. 71Issue 2
- PreviewAlthough vascularised composite allotransplantation (VCA) is increasingly being performed and for a widening range of indications – some now consider transplantation to be the standard of care for mid-forearm amputations2 – more widespread application is limited by the risks associated with lifelong immunosuppression.