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Molecular Medicine

, Volume 8, Issue 9, pp 509–520 | Cite as

Role of Macrophage Migration Inhibitory Factor (MIF) in Peripheral Nerve Regeneration: Anti-MIF Antibody Induces Delay of Nerve Regeneration and the Apoptosis of Schwann Cells

  • Yasuhiko Nishio
  • Jun Nishihira
  • Teruo Ishibashi
  • Hiroyuki Kato
  • Akio Minami
Original Articles

Abstract

Background

Macrophage migration inhibitory factor (MIF) is a pluripotent cytokine involved in inflammation and immune responses as well as in cell growth. Although we previously demonstrated the presence of MIF in peripheral nerves, and MIF mRNA expression was up-regulated after axotomy, the role of MIF in nerve injury and regeneration has not been evaluated.

Materials and Methods

To examine the potential role of MIF in nerve regeneration, we locally administered an anti-MIF polyclonal antibody into regenerating rat sciatic nerves using the silicone chamber model. The effect of the anti-MIF antibody on nerve regeneration was evaluated using an axonal reflex test. In addition, we carried out a terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) assay and immunohistochemical analysis of the damaged nerve segments with regard to apoptosis-related proteins such as p53 to evaluate the effects of anti-MIF antibodies on apoptosis during the regeneration process.

Results

The regeneration length of the nerve in the anti-MIF antibody-treated group was significantly shorter than that in the non-immune rabbit IgG-treated group at weeks 2, 4 and 6 after surgery. TUNEL assay showed that a large number of apoptotic cells, mostly Schwann cells, were observed in the intratubal and distal nerve segments at weeks 4 and 6 after surgery by the anti-MIF antibody treatment. Consistent with these results, Ki-67-positive cells were significantly decreased by the anti-MIF antibody treatment. Immunohistochemical analyses revealed that p53 and, to a lesser extent, Fas were more up-regulated in the anti-MIF antibody-treated nerves than in the controls.

Conclusion

Taken together, these results suggest that MIF plays an important role in acceleration of peripheral nerve regeneration and in prevention of Schwann cell apoptosis, mainly through overcoming the apoptotic effect of p53.

Notes

Acknowledgments

We are grateful to S. Tone for her technical assistance, and to Y. Mizue, of the Sapporo Immunodiagnostic Laboratory, for preparing the anti-rat MIF antibody.

References

  1. 1.
    Holmes W, Young JZ. (1942) Nerve regeneration after immediate and delayed suture. J Anat 77: 63–96.PubMedPubMedCentralGoogle Scholar
  2. 2.
    Perry VH, Brown MC. (1992) Role of macrophages in peripheral nerve degeneration and repair. Bioessays 14: 401–406.CrossRefPubMedGoogle Scholar
  3. 3.
    David JR. (1966) Delayed hypersensitivity in vitro: its mediation by cell free substances forms by lymphoid cell-antigen interaction. Proc. Natl. Acad. Sci. USA 56: 72–77.CrossRefPubMedGoogle Scholar
  4. 4.
    Bloom BR, Bennett B. (1966) Mechanism of a reaction in vitro associated with delayed-type hypersensitivity. Science 153: 80–82.CrossRefGoogle Scholar
  5. 5.
    Bernhagen J, Calandra T, Mitchell RA, et al. (1993) MIF is a pituitary-derived cytokine that potentiates lethal endotoxemia. Nature 365: 756–759.CrossRefPubMedGoogle Scholar
  6. 6.
    Calandra T, Bernhagen J, Metz CN, et al. (1995) MIF as a glucocorticoid-induced modulator of cytokine production. Nature 377: 68–71.CrossRefPubMedGoogle Scholar
  7. 7.
    Bacher M, Metz CN, Calandra T, et al. (1996) An essential regulatory role for macrophage migration inhibitory factor in T-cell activation. Proc. Natl. Acad. Sci. USA 93: 7849–7854.CrossRefPubMedGoogle Scholar
  8. 8.
    Calandra T, Bernhagen J, Mitchell RA, Bucala R. (1994) The macrophage is an important and previously unrecognized source of macrophage migration inhibitory factor. J. Exp. Med. 179: 1895–1902.CrossRefPubMedGoogle Scholar
  9. 9.
    Bucala R. (1996) MIF re-discovered: pituitary hormone and glucocorticoid-induced regulator of cytokine production. FASEB J. 7: 19–24.Google Scholar
  10. 10.
    Wistow GJ, Shaughnessy MP, Lee DC, et al. (1993) A macrophage migration inhibitory factor is expressed in the differentiating cells of the eye lens. Proc. Natl. Acad. Sci. USA 90: 1272–1275.CrossRefPubMedGoogle Scholar
  11. 11.
    Suzuki H, Kanagawa H, Nishihira J. (1996) Evidence for the presence of macrophage migration inhibitory factor in murine reproductive organs and early embryos. Immunol. Lett. 51: 141–147.CrossRefPubMedGoogle Scholar
  12. 12.
    Mitchell RA, Metz CN, Peng T, Bucala, R. (1999) Sustained mitogen-activated protein kinase (MAPK) and cytoplasmic phospholipase A2 activation by macrophage migration inhibitory factor (MIF): regulatory role in cell proliferation and glucocorticoid action. J. Biol. Chem. 274: 18100–18106.CrossRefPubMedGoogle Scholar
  13. 13.
    Chesney J, Metz C, Bacher M, et al. (1999) An essential role for macrophage migration inhibitory factor (MIF) in angiogenesis and the growth of a murine lymphoma. Mol. Med. 5: 181–191.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Shimizu T, Abe R, Nakamura H, et al. (1999) High expression of macrophage migration inhibitory factor in human melanoma cells and its role in tumor cell growth and angiogenesis. Biochem. Biophys. Res. Commun. 264: 751–758.CrossRefPubMedGoogle Scholar
  15. 15.
    Ogawa H, Nishihira J, Sato Y, et al. (2000) An antibody for macrophage migration inhibitory factor suppresses tumor growth and inhibits tumor-associated angiogenesis. Cytokine 12: 309–314.CrossRefPubMedGoogle Scholar
  16. 16.
    Abe R, Shimizu T, Ohkawara A, Nishihira J. (2000) Enhancement of macrophage migration inhibitory factor (MIF) expression in injured epidermis and cultured fibroblasts. Biochim. Biophys. Acta 1500: 1–9.CrossRefPubMedGoogle Scholar
  17. 17.
    Matsuda A, Tagawa Y, Matsuda H, Nishihira J. (1997) Expression of macrophage migration inhibitory factor in corneal wound healing in rats. Invest. Ophthalmol. Vis. Sci. 38: 1555–1562.PubMedGoogle Scholar
  18. 18.
    Nishio Y, Minami A, Kato H, et al. (1999) Identification of macrophage migration inhibitory factor (MIF) in rat peripheral nerves: its possible involvement in nerve regeneration. Biochim. Biophys. Acta 1453: 74–82.CrossRefPubMedGoogle Scholar
  19. 19.
    Nishihira J, Kuriyama T, Sakai M, et al. (1995) The structure and physicochemical properties of rat liver macrophage migration inhibitory factor. Biochim. Biophys. Acta 1247: 159–162.CrossRefPubMedGoogle Scholar
  20. 20.
    Kanje M, Lundborg G, Edstrom A. (1988) A new method for studies of the effects of locally applied drugs on peripheral nerve regeneration in vivo. Brain Res. 439: 116–121.CrossRefPubMedGoogle Scholar
  21. 21.
    Kerns JM, Danielsen N, Holmquist B, et al. (1993) The influence of predegeneration on regeneration through peripheral nerve grafts in the rat. Exp. Neurol. 122: 28–36.CrossRefPubMedGoogle Scholar
  22. 22.
    Danielsson P, Dahlin L, Povlsen B. (1996) Tubulization increases axonal outgrowth of rat sciatic nerve after crush injury. Exp. Neurol. 139: 238–243.CrossRefPubMedGoogle Scholar
  23. 23.
    Ferri CC, Bisby MA. (1999) Improved survival of injured sciatic nerve Schwann cells in mice lacking the p75 receptor. Neurosci. Lett. 272: 191–194.CrossRefPubMedGoogle Scholar
  24. 24.
    Williams LR, Longo FM, Powell HC, et al. (1983) Spatial-temporal progress of peripheral nerve regeneration within a silicone chamber: parameters for a bioassay. J. Comp. Neurol. 218: 460–470.CrossRefPubMedGoogle Scholar
  25. 25.
    Korompilias AV, Chen LE, Seaber AV, Urbaniak JR. (1999) Interleukin-1 beta promotes functional recovery of crushed peripheral nerve. J. Orthop. Res. 17: 714–719.CrossRefPubMedGoogle Scholar
  26. 26.
    Bolin LM, Verity AN, Silver JE, et al. (1995) Interleukin-6 production by Schwann cells and induction in sciatic nerve injury. J. Neurochem. 64: 850–858.CrossRefPubMedGoogle Scholar
  27. 27.
    Bourde O, Kiefer R, Toyka KV, Hartung HP. (1996) Quantification of interleukin-6 mRNA in wallerian degeneration by competitive reverse transcription polymerase chain reaction. J. Neuroimmunol. 69: 135–140.PubMedGoogle Scholar
  28. 28.
    Chen LE, Seaber AV, Wong GH, Urbaniak JR. (1996) Tumor necrosis factor promotes motor functional recovery in crushed peripheral nerve. Neurochem. Int. 29: 197–203.CrossRefPubMedGoogle Scholar
  29. 29.
    Wagner R, Myers RR. (1996) Schwann cells produce tumor necrosis factor alpha: expression in injured and non-injured nerves. Neuroscience 73: 625–629.CrossRefPubMedGoogle Scholar
  30. 30.
    Lindholm D, Heumann R, Meyer M, Thoenen H. (1987) Interleukin-1 regulates synthesis of nerve growth factor in non-neuronal cells of rat sciatic nerve. Nature 330: 658–659.CrossRefPubMedGoogle Scholar
  31. 31.
    Rotshenker S, Aamar S, Barak V. (1992) Interleukin-1 activity in lesioned peripheral nerve. J. Neuroimmuno. l 39: 75–80.CrossRefGoogle Scholar
  32. 32.
    Grinspan JB, Marchionni MA, Reeves M, et al. (1996) Axonal interactions regulate Schwann cell apoptosis in developing peripheral nerve: neuregulin receptors and the role of neuregulins. J. Neurosci. 16: 6107–6118.CrossRefPubMedGoogle Scholar
  33. 33.
    Hudson JD, Shoaibi MA, Maestro R, et al. (1999) A proinflammatory cytokine inhibits p53 tumor suppressor activity. J. Exp. Med. 190: 1375–1382.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Mitchell RA, Liao H, Chesney J, et al. (2002) Macrophage migration inhibitory factor (MIF) sustains macrophage proinflammatory function by inhibiting p53: regulatory role in the innate immune response. Proc. Natl. Acad. Sci. USA 99: 345–350.CrossRefPubMedGoogle Scholar
  35. 35.
    Carter BD, Kaltschmidt C, Kaltschmidt B, et al. (1996) Selective activation of NF-kB by nerve growth factor through the neurotrophin receptor p75. Science 272: 542–545.CrossRefPubMedGoogle Scholar
  36. 36.
    Syroid DE, Maycox PR, Burrola PG, et al. (1996) Cell death in the Schwann cell lineage and its regulation by neuregulin. Proc. Natl. Acad. Sci. USA 93: 9229–9234.CrossRefPubMedGoogle Scholar
  37. 37.
    You S, Petrov T, Chung PH, Gordon T. (1997) The expression of the low affinity nerve growth factor receptor in long-term denervated Schwann cells. Glia 20: 87–100.CrossRefPubMedGoogle Scholar

Copyright information

© NSLIJ Research Institute 2002

Authors and Affiliations

  • Yasuhiko Nishio
    • 1
  • Jun Nishihira
    • 2
  • Teruo Ishibashi
    • 2
  • Hiroyuki Kato
    • 1
  • Akio Minami
    • 1
  1. 1.Department of Orthopaedic SurgeryHokkaido University Graduate School of MedicineSapporoJapan
  2. 2.Department of Molecular BiochemistryHokkaido University Graduate School of MedicineSapporoJapan

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