Over-expression of heme oxygenase-1 does not protect porcine endothelial cells from human xenoantibodies and complement-mediated lysis

  • Chi Zhang (张 弛)
  • Lu Wang (王 璐)
  • Shan Zhong (钟 山)
  • Xiao-xiao Wang (王筱啸)
  • Ying Xiang (向 莹)
  • Shi Chen (陈 实)
  • Gang Chen (陈 刚)
Article

Summary

Accommodated organs can survive in the presence of anti-organ antibodies and complement. Heme oxygenase-1 (HO-1) is essential to ensure accommodation in concordant xenotransplant models. However, whether induction of HO-1 over-expression could protect porcine endothelial cells (PECs) against human xenoantibodies and complement-mediated lysis and induce an in vitro accommodation is still unknown. The SV40-immortalized porcine aorta-derived endothelial cell line (iPEC) was pre-incubated with 20, 50, or 80 μmol/L of cobalt-protoporphyrins IX (CoPPIX) for 24 h, and the HO-1 expression in iPECs was analyzed by using Western blotting. CoPPIX-treated or untreated iPECs were incubated with normal human AB sera, and complement-dependent cytotoxicity (CDC) was measured by both flow cytometry and lactate dehydrogenase (LDH) release assay. In vitro treatment with CoPPIX significantly increased the expression of HO-1 in iPECs in a dose-dependent manner. Over-expression of HO-1 was successfully achieved by incubation of iPECs with either 50 or 80 μmol/L of CoPPIX. However, HO-1 over-expression did not show any protective effects on iPECs against normal human sera-mediated cell lysis. In conclusion, induction of HO-1 over-expression alone is not enough to protect PECs from human xenoantibodies and complement-mediated humoral injury. Additionally, use of other protective strategies is needed to achieve accommodation in pig-to-primate xenotransplantation.

Key words

heme oxygenase-1 porcine endothelial cells complement-dependent cytotoxicity xenotransplantation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Chauveau C, Remy S, Royer PJ, et al. Heme oxygenase-1 expression inhibits dendritic cell maturation and proinflammatory function but conserves IL-10 expression. Blood, 2005,106 (5):1694–1702PubMedCrossRefGoogle Scholar
  2. 2.
    Kinderlerer AR, Pombo Gregoire I, Hamdulay SS, et al. Heme oxygenase-1 expression enhances vascular endothelial resistance to complement-mediated injury through induction of decay-accelerating factor: a role for increased bilirubin and ferritin. Blood, 2009,113(7): 1598–1607PubMedCrossRefGoogle Scholar
  3. 3.
    Duckers HJ, Boehm M, True AL, et al. Heme oxygenase-1 protects against vascular constriction and proliferation. Nat Med, 2001,7(6):693–698PubMedCrossRefGoogle Scholar
  4. 4.
    Soares MP, Lin Y, Sato K, et al. Accommodation. Immunol Today, 1999,20(10):434–437PubMedCrossRefGoogle Scholar
  5. 5.
    Soares MP, Lin Y, Anrather J, et al. Expression of heme oxygenase-1 can determine cardiac xenograft survival. Nat Med, 1998,4(9):1073–1077PubMedCrossRefGoogle Scholar
  6. 6.
    Dorling A, Stocker C, Tsao T, et al. In vitro accommodation of immortalized porcine endothelial cells: resistance to complement-mediated lysis and down-regulation of VCAM expression induced by low concentrations of polyclonal human IgG antipig antibodies. Transplantation, 1996,62(8):1127–1136PubMedCrossRefGoogle Scholar
  7. 7.
    Dalmasso AP, He T, Benson BA. Inhibition of complement-mediated porcine endothelial cell cytotoxicity by human IgM natural antibody. Transplant Proc, 1996,28(2):535PubMedGoogle Scholar
  8. 8.
    Zhu M, Zhang W, Liu F, et al. Resistance to anti-xenogeneic response by combining alpha-Gal silencing with HO-1 upregulation. Transpl Immunol, 2008,19(3–4):202–208PubMedCrossRefGoogle Scholar
  9. 9.
    Chen G, Sun H, Yang H, et al. The role of anti-non-Gal antibodies in the development of acute humoral xenograft rejection of hDAF transgenic porcine kidneys in baboons receiving anti-Gal antibody neutralization therapy. Transplantation, 2006,81(2):273–283PubMedCrossRefGoogle Scholar
  10. 10.
    Rozman P, Kosir A, Bohinjec M. Is the ABO incompatibility a risk factor in bone marrow transplantation? Transpl Immunol, 2005,14(3–4):159–169PubMedCrossRefGoogle Scholar
  11. 11.
    Takahashi K, Saito K, Takahara S, et al. Excellent long-term outcome of ABO-incompatible living donor kidney transplantation in Japan. Am J Transplant, 2004, 4(7):1089–1096PubMedCrossRefGoogle Scholar
  12. 12.
    Hanto DW, Fecteau AH, Alonso MH, et al. ABO-incompatible liver transplantation with no immunological graft losses using total plasma exchange, splenectomy, and quadruple immunosuppression: evidence for accommodation. Liver Transpl, 2003,9(1): 22–30PubMedCrossRefGoogle Scholar
  13. 13.
    Sato K, Balla J, Otterbein L, et al. Carbon monoxide generated by heme oxygenase-1 suppresses the rejection of mouse-to-rat cardiac transplants. J Immunol, 2001,166(6):4185–4194PubMedGoogle Scholar
  14. 14.
    Wang N, Lee JM, Tobiasch E, et al. Induction of xenograft accommodation by modulation of elicited antibody responses1 2. Transplantation, 2002,74(3): 334–445PubMedCrossRefGoogle Scholar
  15. 15.
    Salama AD, Delikouras A, Pusey CD, et al. Transplant accommodation in highly sensitized patients: a potential role for Bcl-xL and alloantibody. Am J Transplant, 2001,1(3):260–269PubMedCrossRefGoogle Scholar
  16. 16.
    Tu CF, Tai HC, Wu CP, et al. The in vitro protection of human decay accelerating factor and hDAF/heme oxygenase-1 transgenes in porcine aortic endothelial cells against sera of Formosan macaques. Transplant Proc, 2010,42(6):2138–2141PubMedCrossRefGoogle Scholar

Copyright information

© Huazhong University of Science and Technology and Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Chi Zhang (张 弛)
    • 1
    • 4
  • Lu Wang (王 璐)
    • 1
  • Shan Zhong (钟 山)
    • 1
  • Xiao-xiao Wang (王筱啸)
    • 1
  • Ying Xiang (向 莹)
    • 1
  • Shi Chen (陈 实)
    • 1
    • 2
    • 3
  • Gang Chen (陈 刚)
    • 1
    • 2
    • 3
  1. 1.Institute of Organ TransplantationHuazhong University of Science and TechnologyWuhanChina
  2. 2.Key Laboratory of Organ Transplantation, Ministry of EducationHuazhong University of Science and TechnologyWuhanChina
  3. 3.Key Laboratory of Organ Transplantation, Ministry of Public Health, Tongji Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  4. 4.Department of UrologyFujian Provincial HospitalFujianChina

Personalised recommendations