title: Transgenic expression of human cytoxic T-lymphocyte associated antigen4-immunoglobulin (hCTLA4Ig) by porcine skin for xenogeneic skin grafting.
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.
authors.affiliation: Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Shapingba District, Chongqing, 400038, China, yongw7528@gmail.com.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Hua-Qiang
authors.initials: HQ
authors.lastname: Yang
authors.forename: Wen
authors.initials: W
authors.lastname: Jiang
authors.forename: Na-Na
authors.initials: NN
authors.lastname: Fan
authors.forename: Ben-Tian
authors.initials: BT
authors.lastname: Zhao
authors.forename: Zhen
authors.initials: Z
authors.lastname: Ou-Yang
authors.forename: Zhao-Ming
authors.initials: ZM
authors.lastname: Liu
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhao
authors.forename: Dong-Shan
authors.initials: DS
authors.lastname: Yang
authors.forename: Xiao-Yang
authors.initials: XY
authors.lastname: Zhou
authors.forename: Hai-Tao
authors.initials: HT
authors.lastname: Shang
authors.forename: Lu-Lu
authors.initials: LL
authors.lastname: Wang
authors.forename: Peng-Ying
authors.initials: PY
authors.lastname: Xiang
authors.forename: Liang-Peng
authors.initials: LP
authors.lastname: Ge
authors.forename: Hong
authors.initials: H
authors.lastname: Wei
authors.forename: Liang-Xue
authors.initials: LX
authors.lastname: Lai
mesh_terms: D000069594:Abatacept / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D006085:Graft Survival; D006801:Humans; D007633:Keratins / Q000235:genetics; D051379:Mice; D053652:Nuclear Transfer Techniques; D011401:Promoter Regions, Genetic; D051381:Rats; D016038:Skin Transplantation; D013552:Swine / Q000235:genetics; D014183:Transplantation, Heterologous
chemical_list: D007633:Keratins; D000069594:Abatacept