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Transgenic expression of human cytoxic T-lymphocyte associated antigen4-Immunoglobulin (hCTLA4Ig) by porcine skin for xenogeneic skin grafting

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Abstract

Porcine skin is frequently used as a substitute of human skin to cover large wounds in clinic practice of wound care. In our previous work, we found that transgenic expression of human cytoxicT-lymphocyte associated antigen4-immunoglobulin (hCTLA4Ig) in murine skin graft remarkably prolonged its survival in xenogeneic wounds without extensive immunosuppression in recipients, suggesting that transgenic hCTLA4Ig expression in skin graft may be an effective and safe method to prolong xenogeneic skin graft survival. In this work, using a transgene construct containing hCTLA4Ig coding sequence under the drive of human Keratine 14 (k14) promoter, hCTLA4Ig transgenic pigs were generated by somatic nuclear transfer. The derived transgenic pigs were healthy and exhibited no signs of susceptibility to infection. The hCTLA4Ig transgene was stably transmitted through germline over generations, and thereby a transgenic pig colony was established. In the derived transgenic pigs, hCTLA4Ig expression in skin was shown to be genetically stable over generations, and detected in heart, kidney and corneal as well as in skin. Transgenic hCTLA4Ig protein in pigs exhibited expected biological activity as it suppressed human lymphocyte proliferation in human mixed lymphocyte culture to extents comparable to those of commercially purchased purified hCTLA4Ig protein. In skin grafting from pigs to rats, transgenic porcine skin grafts exhibited remarkably prolonged survival compared to the wild-type skin grafts derived from the same pig strain (13.33 ± 3.64 vs. 6.25 ± 2.49 days, P < 0.01), further indicating that the transgenic hCTLA4Ig protein was biologically active and capable of extending porcine skin graft survival in xenogeneic wounds. The transgenic pigs generated in this work can be used as a reproducible resource to provide porcine skin grafts with extended survival for wound coverage, and also as donors to investigate the impacts of hCTLA4Ig on xenotransplantation of other organs (heart, kidney and corneal) due to the ectopic transgenic hCTLA4Ig expression.

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Acknowledgments

This work was supported by grants from National 863 Project of China (2012AA020504), Natural Science Fund of China (NSFC, 31171280), National 973 Project of China (2011CB944102), Chongqing Natural Science Fund(cstc2011jjA10049), International Science and Technology Cooperation Program of China (S2013ZR0398), Advancing Research in Basic Science of Chongqing (cstc2013jcyjC80001) and Chongqing Agriculture Development Fund (12402).

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Authors and Affiliations

  1. Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Shapingba District, Chongqing, 400038, China

    Yong Wang, Wen Jiang, Xiao-Yang Zhou, Hai-Tao Shang, Lu-Lu Wang, Peng-Ying Xiang & Hong Wei

  2. Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, Guangdong, China

    Hua-Qiang Yang, Na-Na Fan, Ben-Tian Zhao, Zhen Ou-Yang, Zhao-Ming Liu, Yu Zhao, Dong-Shan Yang & Liang-Xue Lai

  3. Chongqing Academy of Animal Sciences, Chongqing, 402460, Rongchang County, China

    Liang-Peng Ge

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  1. Yong Wang
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  2. Hua-Qiang Yang
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  3. Wen Jiang
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  12. Lu-Lu Wang
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  14. Liang-Peng Ge
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  15. Hong Wei
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  16. Liang-Xue Lai
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Corresponding authors

Correspondence to Hong Wei or Liang-Xue Lai.

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Yong Wang and Hua-Qiang Yang have contributed equally to this work.

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Wang, Y., Yang, HQ., Jiang, W. et al. Transgenic expression of human cytoxic T-lymphocyte associated antigen4-Immunoglobulin (hCTLA4Ig) by porcine skin for xenogeneic skin grafting. Transgenic Res 24, 199–211 (2015). https://doi.org/10.1007/s11248-014-9833-9

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  • DOI: https://doi.org/10.1007/s11248-014-9833-9

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