Differentiation Between Vascularized Composite Allograft Acute Skin Rejection and Delayed Type Hypersensitivity Reactions Based on Cytokine Analysis

Background: Skin rejection in vascularized composite allotransplantation is a major hurdle for wider clinical adoption in VCA. Histopathological alterations during acute skin rejection mimic inflammatory skin processes and are difficult to differentiate. The aim of this study was to identify protein expression patterns characteristic for acute skin rejection in reference to inflammatory skin processes, such as delayed type hypersensitivity reactions. Methods: Limb transplantations were performed using a Brown-Norway to Lewis rat allotransplant model. For the induction of skin inflammation a contact hypersensitivity (CHS) and delayed type hypersensitivity (DTH) model in Lewis rats were established. CHS and DTH result in skin irritation and swelling comparable to grade I-II rejection in our rat allotransplant model. While the DTH reaction is limited to the plantar skin, the CHS model allows for comparison with rejection also in hairy skin. Skin biopsies were taken at defined time points, where a histologically identical inflammatory response in all study groups was present. Protein levels of 14 inflammatory cytokines were assessed by Luminex™. Results: Based on the multivariate linear discriminant analysis, IL-12p70 and TNF-α were the major discriminators between skin rejection and inflammatory skin reactions. Principal component analysis identified IL-1α, GM-CSF, IL-6 and IL-18 as key drivers of rejection at this time point. Conclusion: The novel diagnostic platform enables for early and specific diagnosis of rejection and differentiation toward inflammatory skin diseases in this transplant model.

[1]  G. Pierer,et al.  Insights from Computational Modeling in Inflammation and Acute Rejection in Limb Transplantation , 2014, PloS one.

[2]  P. Ye,et al.  Anti‐interleukin‐12/23p40 antibody attenuates chronic rejection of cardiac allografts partly via inhibition γδT cells , 2012, Clinical and experimental immunology.

[3]  Honghui Zhou,et al.  Therapeutic targeting of the IL-12/23 pathways: generation and characterization of ustekinumab , 2011, Nature Biotechnology.

[4]  P. Ye,et al.  Treatment With Interleukin-12/23p40 Antibody Attenuates Acute Cardiac Allograft Rejection , 2011, Transplantation.

[5]  R. Margreiter,et al.  Molecular Markers and Targeted Therapy of Skin Rejection in Composite Tissue Allotransplantation , 2010, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[6]  Jean Kanitakis.  The challenge of dermatopathological diagnosis of composite tissue allograft rejection: a review , 2008, Journal of cutaneous pathology.

[7]  A. Kirk,et al.  The Banff 2007 Working Classification of Skin‐Containing Composite Tissue Allograft Pathology , 2008, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[8]  F. Schuind,et al.  Atypical Acute Rejection After Hand Transplantation , 2008, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[9]  S. Nakae,et al.  IL-1beta, but not IL-1alpha, is required for antigen-specific T cell activation and the induction of local inflammation in the delayed-type hypersensitivity responses. , 2006, International immunology.

[10]  P. Tak,et al.  Pharmacokinetics of IL-18 binding protein in healthy volunteers and subjects with rheumatoid arthritis or plaque psoriasis , 2006, European Journal of Drug Metabolism and Pharmacokinetics.

[11]  J. Kong,et al.  Regulation of IL-1 Family Cytokines IL-1α, IL-1 Receptor Antagonist, and IL-18 by 1,25-Dihydroxyvitamin D3 in Primary Keratinocytes1 , 2006, The Journal of Immunology.

[12]  F. Bérard,et al.  The role of CD4+ and CD8+ T cells in contact hypersensitivity and allergic contact dermatitis. , 2004, European journal of dermatology : EJD.

[13]  A. Mamelak,et al.  Cytokine knockouts in contact hypersensitivity research. , 2003, Cytokine & growth factor reviews.

[14]  M. Gerstein,et al.  Comparing protein abundance and mRNA expression levels on a genomic scale , 2003, Genome Biology.

[15]  G. Burmester,et al.  Anakinra, a recombinant human interleukin-1 receptor antagonist (r-metHuIL-1ra), in patients with rheumatoid arthritis: A large, international, multicenter, placebo-controlled trial. , 2003, Arthritis and rheumatism.

[16]  R. Groves,et al.  Increased migration of murine keratinocytes under hypoxia is mediated by induction of urokinase plasminogen activator. , 2002, The Journal of investigative dermatology.

[17]  D. Sauder,et al.  Contribution of Langerhans Cell-Derived IL-18 to Contact Hypersensitivity , 2002, The Journal of Immunology.

[18]  S. Nakae,et al.  IL-1α, but not IL-1β, is required for contact-allergen-specific T cell activation during the sensitization phase in contact hypersensitivity , 2001 .

[19]  Y. Ohta,et al.  Expression of IL-18 in psoriasis , 2001, Archives of Dermatological Research.

[20]  Jean Kanitakis,et al.  Sequential histological and immunohistochemical study of the skin of the first human hand allograft. , 2000, Transplantation.

[21]  A. Zipfel,et al.  TNF-α and Its Receptors Mediate Graft Rejection and Loss after Liver Transplantation , 2000 .

[22]  R. Karr,et al.  Mice deficient in IL-1beta manifest impaired contact hypersensitivity to trinitrochlorobenzone , 1996, The Journal of experimental medicine.

[23]  S. Kondo,et al.  Interleukin-1 receptor antagonist suppresses contact hypersensitivity. , 1995, The Journal of investigative dermatology.

[24]  L. Bubnova,et al.  Interleukin-1 beta and tumor necrosis factor-alpha serum levels in renal allograft recipients. , 1992, Transplantation proceedings.

[25]  G. Trinchieri,et al.  Human dermal mast cells contain and release tumor necrosis factor alpha, which induces endothelial leukocyte adhesion molecule 1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. Chia,et al.  The role of tumor necrosis factor in allograft rejection. I. Evidence that elevated levels of tumor necrosis factor-alpha predict rejection following orthotopic liver transplantation. , 1990, Transplantation.

[27]  S. Nakae,et al.  IL-1 alpha, but not IL-1 beta, is required for contact-allergen-specific T cell activation during the sensitization phase in contact hypersensitivity. , 2001, International immunology.

[28]  A. Zipfel,et al.  TNF-alpha and its receptors mediate graft rejection and loss after liver transplantation. , 2000, Clinical chemistry and laboratory medicine.

[29]  C. Dinarello.  IL-18: A TH1-inducing, proinflammatory cytokine and new member of the IL-1 family. , 1999, The Journal of allergy and clinical immunology.

[30]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[31]  A. Gaber,et al.  Elevated tumor necrosis factor levels are predictive for pancreas allograft transplant rejection. , 1993, Transplantation proceedings.