Generation of soluble human tumor necrosis factor-α receptor 1-Fc transgenic pig

Bumrae Cho, Ok Jae Koo, Jong-Ik Hwang, Hwajung Kim, Eun Mi Lee, Sunghoon Hurh, Sol Ji Park, Han Ro, Jaeseok Yang, Charles D. Surh, Anthony J. D'Apice, Byeong Chun Lee, Curie Ahn

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23 Citations (Scopus)

Abstract

Background. Acute humoral xenograft rejection (AHXR) is an important barrier to xenograft survival. Human tumor necrosis factor-α (hTNF-α) is one of the essential mediators of AHXR and induces activation of porcine endothelial cells (PECs), resulting in upregulation of major histocompatibility complex molecules, adhesion molecules, and proinflammatory chemokines. We investigated whether introduction of a soluble human tumor necrosis factor receptor I-Fc (shTNFRI-Fc) fusion gene can suppress activation of PECs and, more importantly, produced shTNFRI-Fc transgenic pigs. Methods. The shTNFRI-Fc gene expression vector was constructed and inserted into PECs. The inhibitory effects of shTNFRI-Fc were tested by luciferase assay, reverse-transcriptase polymerase chain reaction, and flow cytometry. A shTNFRI-Fc transgenic pig was generated by somatic cell nuclear transfer. The expression of shTNFRI-Fc in the transgenic pig was evaluated by PCR, western blot, enzyme-linked immunosorbent assay, and immunohistochemistry. The inhibitory effects of shTNFRI-Fc in the serum obtained from the transgenic pig were also tested. Results. In comparison with control green fluorescent protein, shTNFRI-Fc protein showed much stronger inhibitory effects on NF-κB activation in the HEK293-NF-κB-luciferase reporting cell line, expression of chemokines and adhesion molecules in PECs, and TNF-α-mediated cytotoxicity. We successfully generated shTNFRI-Fc transgenic pig. Sera obtained from the transgenic pig inhibited induction of chemokines, and E-selectin in PECs stimulated with Human TNF-α. CONCLUSIONS.: We have generated transgenic pigs producing shTNFRI-Fc protein that can inhibit TNF-α-mediated activation of PECs. Because TNF-α is an important mediator of xenograft rejection, the use of xenografts that can produce shTNFRI-Fc proteins de novo could be an effective approach in overcoming a considerable component of the xenograft rejection process, especially AHXR.

Original languageEnglish
Pages (from-to)139-147
Number of pages9
JournalTransplantation
Volume92
Issue number2
DOIs
Publication statusPublished - 2011 Jul 27

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Keywords

  • Acute humoral xenograft rejection
  • shTNFRI-Fc
  • TNF-α
  • Transgenic pig
  • Xenotransplantation

ASJC Scopus subject areas

  • Transplantation

Cite this

Cho, B., Koo, O. J., Hwang, J-I., Kim, H., Lee, E. M., Hurh, S., Park, S. J., Ro, H., Yang, J., Surh, C. D., D'Apice, A. J., Lee, B. C., & Ahn, C. (2011). Generation of soluble human tumor necrosis factor-α receptor 1-Fc transgenic pig. Transplantation, 92(2), 139-147. https://doi.org/10.1097/TP.0b013e3182215e7e