Peripheral Nerve Transplantation: The Role of Chemical Acellularization in Eliminating Allograft Antigenicity

ABSTRACT

This study tests the hypothesis that a chemically acellularized peripheral nerve allograft is as immunologically inactive as a peripheral nerve isograft. Cellular and acellular sciatic nerves were transplanted from BALB/c into C57BL/6 mice. C57BL/6 sciatic nerves were also transplanted into C57BL/6 recipients as isograft controls. Fourteen days post-transplantation, recipient splenocytes were isolated, stimulated with donor alloantigens, and IL-2, IL-4, IL-5, and γ-IFN production was quantified using the ELISPOT technique. Cellular peripheral nerve allografts stimulated robust Th1 and Th2 systemic immune responses, whereas acellular peripheral nerve allografts elicited a response that is comparable to or lower than that quantified following peripheral nerve isograft transplantation. Chemical acellularization of peripheral nerve allografts dramatically reduces the cellular and humoral immunologic responses. These data indicate that chemically acellularized peripheral nerve constructs are relatively non-antigenic and may be a readily available source of nerve for peripheral nerve reconstruction.

KEYWORDS

Nerve - immunology - transplantation

REFERENCES

• 1 Terzis J K, Sun D D, Thanos P K. Historical and basic science review: past, present, and future of nerve repair.  J Reconstr Microsurg. 1997;  13 215-225
• 2 Watchmaker G P, Mackinnon S E. Advances in peripheral nerve repair.  Clin Plast Surg. 1997;  24 63-73
• 3 Evans P J, Midha R, Mackinnon S E. The peripheral nerve allograft: a comprehensive review of regeneration and neuroimmunology.  Prog Neurobiol. 1994;  43 187-233
• 4 Mackinnon S E, Dellon A L. Clinical nerve reconstruction with a bioabsorbable polyglycolic acid tube.  Plast Reconstr Surg. 1990;  85 419-424
• 5 Weber R A, Breidenbach W C, Brown R E, Jabaley M E, Mass D P. A randomized prospective study of polyglycolic acid conduits for digital nerve reconstruction in humans.  Plast Reconstr Surg. 2000;  106 1036-1045
• 6 Walton R L, Brown R E, Matory Jr W E, Borah G L, Dolph J L. Autogenous vein graft repair of digital nerve defects in the finger: a retrospective clinical study.  Plast Reconstr Surg. 1989;  84 944-949
• 7 Chiu D T, Lovelace R E, Yu L T et al.. Comparative electrophysiologic evaluation of nerve grafts and autogenous vein grafts as nerve conduits: an experimental study.  J Reconstr Microsurg. 1988;  4 303-309
• 8 Bunge M B, Clark M B, Dean A C, Eldridge C F, Bunge R P. Schwann cell function depends upon axonal signals and basal lamina components.  Ann N Y Acad Sci. 1990;  580 281-287
• 9 Rodriguez F J, Verdu E, Ceballos D, Navarro X. Nerve guides seeded with autologous Schwann cells improve nerve regeneration.  Exp Neurol. 2000;  161 571-584
• 10 Sondell M, Lundborg G, Kanje M. Regeneration of the rat sciatic nerve into allografts made acellular through chemical extraction.  Brain Res. 1998;  795 44-54
• 11 Argall K G, Armati P J, Pollard J D, Watson E, Bonner J. Interactions between CD4+ T-cells and rat Schwann cells in vitro. 1. Antigen presentation by Lewis rat Schwann cells to P2-specific CD4+ T- cell lines.  J Neuroimmunol. 1992;  40 1-18
• 12 Kingston A E, Bergsteinsdottir K, Jessen K R, Van der Meide P H, Colston M J, Mirsky R. Schwann cells co-cultured with stimulated T cells and antigen express major histocompatibility complex (MHC) class II determinants without interferon-gamma pretreatment: synergistic effects of interferon-gamma and tumor necrosis factor on MHC class II induction.  Eur J Immunol. 1989;  19 177-183
• 13 Argall K G, Armati P J, Pollard J D, Bonner J. Interactions between CD4+ T-cells and rat Schwann cells in vitro. 2. Cytotoxic effects of P2-specific CD4+ T-cell lines on Lewis rat Schwann cells.  J Neuroimmunol. 1992;  40 19-29
• 14 Stoll G, Muller H W. Nerve injury, axonal degeneration and neural regeneration: basic insights.  Brain Pathol. 1999;  9 313-325
• 15 Funakoshi H, Frisen J, Barbany G et al.. Differential expression of mRNAs for neurotrophins and their receptors after axotomy of the sciatic nerve.  J Cell Biol. 1993;  123 455-465
• 16 Dennis R G, Kuzon W M, Cederna P S. U.S. Serial No. 09/896,651, Method For Chemically Acellularizing a Biological Tissue Sample: Continuation in Part of U.S. Serial No. 09/709,890. U.S Patent No. 6,448,076, September 10. 2002
• 17 Matesic D, Lehmann P V, Heeger P S. High-resolution characterization of cytokine-producing alloreactivity in naive and allograft-primed mice.  Transplantation. 1998;  65 906-914
• 18 Kramer H. The metachromatic staining reaction.  J Histochem. 1955;  3 227-232
• 19 Chan S Y, DeBruyne L A, Goodman R E, Eichwald E J, Bishop D K. In vivo depletion of CD8+ T cells results in Th2 cytokine production and alternate mechanisms of allograft rejection.  Transplantation. 1995;  59 1155-1161
• 20 Bishop D K, Shelby J, Eichwald E J. Mobilization of T lymphocytes following cardiac transplantation: evidence the CD4-positive cells are required for cytotoxic T lymphocyte activation, inflammatory endothelial development, graft infiltration, and acute allograft rejection.  Transplantation. 1992;  53 849-857
• 21 Piccotti J R, Chan S Y, VanBuskirk A M, Eichwald E J, Bishop D. Are Th2 helper T lymphocytes beneficial, deleterious, or irrelevant in promoting allograft survival?.  Transplantation. 1997;  63 619-624
• 22 Bishop D K, ChanWood S, Eichwald E J, Orosz C G. Immunobiology of allograft rejection in the absence of IFNγ: CD8+ effector cells develop independently of CD4+ cells and CD40-CD40 ligand interactions.  J Immunol. 2001;  166 3248-3255
• 23 Bishop D K, Li W, Chan S Y, Ensley R D, Shelby J, Eichwald E J. Helper T lymphocyte unresponsiveness to cardiac allografts following transient depletion of CD4-positive cells.  Implications for cellular and humoral responses. Transplantation. 1994;  58 576-584
• 24 Fu S Y, Gordon T. The cellular and molecular basis of peripheral nerve regeneration.  Mol Neurobiol. 1997;  14 67-116
• 25 Piccotti J R, Chan S Y, Eichwald E J, Bishop D K. Differential effects of IL-12 receptor blockade with IL-12 p40 homodimer on the induction of CD4+ and CD8+ IFN-γ-producing cells.  J Immunol. 1997;  158 643-648
• 26 Krams S M, Falco D A, Villanueva J C et al.. Cytokine and T cell receptor gene expression at the site of allograft rejection.  Transplantation. 1992;  53 151-156
• 27 Martinez O M, Villanueva J C, Lake J, Roberts J P, Ascher N L, Krams S M. IL-2 and IL-5 gene expression in response to alloantigen in liver allograft recipients and in vitro.  Transplantation. 1993;  55 1159-1166
• 28 VanBuskirk A M, Wakely M E, Orosz C G. Transfusion of polarized TH2-like cell populations into SCID mouse cardiac allograft recipients results in acute allograft rejection.  Transplantation. 1996;  62 229-238
• 29 Strom T B, Roy-Chaudhury P, Manfro R et al.. The Th1/Th2 paradigm and the allograft response.  Curr Opin Immunol. 1996;  8 688-693
• 30 Baumgarth N. A two-phase model of B-cell activation.  Immunol Rev. 2000;  176 171-180
• 31 Haase S C, Rovak J M, Dennis R G, Kuzon W M, Cederna P S. Recovery of muscle contractile function following peripheral nerve gap repair with an acellularized peripheral nerve graft.  J Reconstr Microsurg. 2005;  , in press
• 32 Haase S C, Cederna P S, Dennis R G, Kuzon W M. Peripheral nerve reconstruction using acellular nerve grafts.  Surg Forum. 2000;  51 607-609