J Korean Neurosurg Soc.  2015 Aug;58(2):101-106. 10.3340/jkns.2015.58.2.101.

T Lymphocyte Subsets and Cytokines in Rats Transplanted with Adipose-Derived Mesenchymal Stem Cells and Acellular Nerve for Repairing the Nerve Defects

• Jiang LF
• Chen O
• Chu TG
• Ding J
• Yu Q

• KMID: 2191316
• DOI: http://doi.org/10.3340/jkns.2015.58.2.101

Abstract

OBJECTIVE
The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects.
METHODS
ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft.
RESULTS
At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of CD3+, CD4+, and CD8+ subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of CD3+, CD4+, and CD8+ subsets and the serum concentration of IL-2, TNF-alpha, and IFN-gamma in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of CD3+, CD4+, and CD8+ subsets and the serum concentration of IL-2, TNF-alpha, and IFN-gamma were significantly reduced in group C (p<0.05).
CONCLUSION
The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.

Keyword

Neural transplantation; Immunity; T cell subsets; Cytokines

MeSH Terms

Allografts
Animals
Autografts
Cytokines*
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Interferon-gamma
Interleukin-2
Mesenchymal Stromal Cells*
Orbit
Rats*
Rats, Sprague-Dawley
Rats, Wistar
Sciatic Nerve
T-Lymphocyte Subsets*
Transplants
Tumor Necrosis Factor-alpha
Cytokines
Interferon-gamma
Interleukin-2
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 Repairing sciatic nerve defects with artificial nerve.

• 

  Fig. 2 The expression of CD44 (A), CD45 (B), and CD90 (C) of ADSCs determined using flow cytometry. ADSCs : adipose-derived mesenchymal stem cells.

• 

  Fig. 3 Longitudinal section of acellular nerve displayed an intact three-dimensional structure (scanning electron microscopy, 5000×). IL-2 : interleukin-2, TNF-α : tumor necrosis factor-α, IFN-γ : interferon-γ.

• 

  Fig. 4 The proportion of CD3+, CD4+, and CD8+ subsets and the serum concentration of IL-2, IFN-γ, and TNF-α. IL-2 : interleukin-2, TNF-α : tumor necrosis factor-α, IFN-γ : interferon-γ.

Reference

1. Bain JR, Mackinnon SE, Hudson AR, Falk RE, Falk JA, Hunter DA. The peripheral nerve allograft : an assessment of regeneration across nerve allografts in rats immunosuppressed with cyclosporin A. Plast Reconstr Surg. 1988; 82:1052–1066. PMID: 3264409.

2. Comoli P, Ginevri F, Maccario R, Avanzini MA, Marconi M, Groff A, et al. Human mesenchymal stem cells inhibit antibody production induced in vitro by allostimulation. Nephrol Dial Transplant. 2008; 23:1196–1202. PMID: 18029377.

3. de Ruiter GC, Spinner RJ, Yaszemski MJ, Windebank AJ, Malessy MJ. Nerve tubes for peripheral nerve repair. Neurosurg Clin N Am. 2009; 20:91–105. viiPMID: 19064182.

4. Gao X, Wang Y, Chen J, Peng J. The role of peripheral nerve ECM components in the tissue engineering nerve construction. Rev Neurosci. 2013; 24:443–453. PMID: 23907421.

5. Hong SJ, Traktuev DO, March KL. Therapeutic potential of adipose-derived stem cells in vascular growth and tissue repair. Curr Opin Organ Transplant. 2010; 15:86–91. PMID: 19949335.

6. Hudson TW, Zawko S, Deister C, Lundy S, Hu CY, Lee K, et al. Optimized acellular nerve graft is immunologically tolerated and supports regeneration. Tissue Eng. 2004; 10:1641–1651. PMID: 15684673.

7. Jiang L, Zhu JK, Liu XL, Xiang P, Hu J, Yu WH. Differentiation of rat adipose tissue-derived stem cells into Schwann-like cells in vitro. Neuroreport. 2008; 19:1015–1019. PMID: 18580571.

8. Kang HG, Zhang D, Degauque N, Mariat C, Alexopoulos S, Zheng XX. Effects of cyclosporine on transplant tolerance : the role of IL-2. Am J Transplant. 2007; 7:1907–1916. PMID: 17617853.

9. Kingham PJ, Kalbermatten DF, Mahay D, Armstrong SJ, Wiberg M, Terenghi G. Adipose-derived stem cells differentiate into a Schwann cell phenotype and promote neurite outgrowth in vitro. Exp Neurol. 2007; 207:267–274. PMID: 17761164.

10. Klyushnenkova E, Mosca JD, Zernetkina V, Majumdar MK, Beggs KJ, Simonetti DW, et al. T cell responses to allogeneic human mesenchymal stem cells : immunogenicity, tolerance, and suppression. J Biomed Sci. 2005; 12:47–57. PMID: 15864738.

11. Kuo YR, Chen CC, Goto S, Lee IT, Huang CW, Tsai CC, et al. Modulation of immune response and T-cell regulation by donor adipose-derived stem cells in a rodent hind-limb allotransplant model. Plast Reconstr Surg. 2011; 128:661e–672e.

12. Kvist M, Sondell M, Kanje M, Dahlin LB. Regeneration in, and properties of, extracted peripheral nerve allografts and xenografts. J Plast Surg Hand Surg. 2011; 45:122–128. PMID: 21682608.

13. Lin CS, Lin G, Lue TF. Allogeneic and xenogeneic transplantation of adipose-derived stem cells in immunocompetent recipients without immunosuppressants. Stem Cells Dev. 2012; 21:2770–2778. PMID: 22621212.

14. Liu GB, Cheng YX, Feng YK, Pang CJ, Li Q, Wang Y, et al. Adipose-derived stem cells promote peripheral nerve repair. Arch Med Sci. 2011; 7:592–596. PMID: 22291793.

15. Mackinnon SE, Doolabh VB, Novak CB, Trulock EP. Clinical outcome following nerve allograft transplantation. Plast Reconstr Surg. 2001; 107:1419–1429. PMID: 11335811.

16. Marconi S, Castiglione G, Turano E, Bissolotti G, Angiari S, Farinazzo A, et al. Human adipose-derived mesenchymal stem cells systemically injected promote peripheral nerve regeneration in the mouse model of sciatic crush. Tissue Eng Part A. 2012; 18:1264–1272. PMID: 22332955.

17. Rovak JM, Bishop DK, Boxer LK, Wood SC, Mungara AK, Cederna PS. Peripheral nerve transplantation : the role of chemical acellularization in eliminating allograft antigenicity. J Reconstr Microsurg. 2005; 21:207–213. PMID: 15880301.

18. Santiago LY, Clavijo-Alvarez J, Brayfield C, Rubin JP, Marra KG. Delivery of adipose-derived precursor cells for peripheral nerve repair. Cell Transplant. 2009; 18:145–158. PMID: 19499703.

19. Sioud M, Mobergslien A, Boudabous A, Fløisand Y. Evidence for the involvement of galectin-3 in mesenchymal stem cell suppression of allogeneic T-cell proliferation. Scand J Immunol. 2010; 71:267–274. PMID: 20384870.

20. 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. PMID: 9622591.

21. Sotiropoulou PA, Perez SA, Gritzapis AD, Baxevanis CN, Papamichail M. Interactions between human mesenchymal stem cells and natural killer cells. Stem Cells. 2006; 24:74–85. PMID: 16099998.

22. Sun XH, Che YQ, Tong XJ, Zhang LX, Feng Y, Xu AH, et al. Improving nerve regeneration of acellular nerve allografts seeded with SCs bridging the sciatic nerve defects of rat. Cell Mol Neurobiol. 2009; 29:347–353. PMID: 18987968.

23. Tung TH, Mohanakumar T, Mackinnon SE. TH1/TH2 cytokine profile of the immune response in limb component transplantation. Plast Reconstr Surg. 2005; 116:557–566. PMID: 16079692.

24. Velásquez SY, García LF, Opelz G, Alvarez CM, Süsal C. Release of soluble CD30 after allogeneic stimulation is mediated by memory T cells and regulated by IFN-γ and IL-2. Transplantation. 2013; 96:154–161. PMID: 23857000.

25. Wang Y, Zhao Z, Ren Z, Zhao B, Zhang L, Chen J, et al. Recellularized nerve allografts with differentiated mesenchymal stem cells promote peripheral nerve regeneration. Neurosci Lett. 2012; 514:96–101. PMID: 22405891.

26. Yu X, Jiang Y, Lu L, Gong X, Sun X, Xuan Z, et al. A crucial role of IL-17 and IFN-γ during acute rejection of peripheral nerve xenotransplantation in mice. PLoS One. 2012; 7:e34419. PMID: 22479627.

27. Zhang Y, Luo H, Zhang Z, Lu Y, Huang X, Yang L, et al. A nerve graft constructed with xenogeneic acellular nerve matrix and autologous adipose-derived mesenchymal stem cells. Biomaterials. 2010; 31:5312–5324. PMID: 20381139.

28. Zimmerer JM, Horne PH, Fiessinger LA, Fisher MG, Pham TA, Saklayen SL, et al. Cytotoxic effector function of CD4-independent, CD8(+) T cells is mediated by TNF-α/TNFR. Transplantation. 2012; 94:1103–1110. PMID: 23222736.