Skin-specifically transgenic expression of biologically active human cytoxic T-lymphocyte associated Antigen4-Immunoglobulin (hCTLA4Ig) in mice using lentiviral vector

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• 作者：Wen Jiang (1)
  Xiao-yang Zhou (1)
  Lu-lu Wang (1)
  Qin Liu (1)
  Cang’e Liu (1)
  Yong Wang (1) yongw7528@gmail.com
  Hong Wei (1) weihong63528@163.com
• 关键词：Lentiviral vector – Transgenic – hCTLA4Ig – Skin grafting
• 刊名：Transgenic Research
• 出版年：2012
• 出版时间：June 2012
• 年：2012
• 卷：21
• 期：3
• 页码：579-591
• 全文大小：595.9 KB
• 参考文献：1. Desai MH, Herndon DN, Broemeling L et al (1990) Early burn wound excision significantly reduces blood loss. Ann Surg 211:753–759
  2. Guerder S, Flavell RA (1995) Co-stimulation in immune tolerance and autoimmunity. Int Rev Immunol 1995(13):135–146
  3. Guo L, Fujino M, Kimura H, Funeshima N, Kitazawa Y, Harihara Y, Tezuka K, Makuuchi M, Suzuki S, Li XK (2003) Simultaneous blockade of co-stimulatory signals, CD28 and ICOS, induced a stable tolerance in rat heart transplantation. Transpl Immunol 12(1):41–48
  4. Hofmann A, Kessler B, Ewerling S et al (2003) Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep 4:1054–1060
  5. Hogan B, Beddington R, Costantini F, Lacy B (1994) Manipulating the mouse embryo: a laboratory manual, second edition. Cold Spring Harbor Laboratory Press
  6. Jin YZ, Xie SS (2003) Bicistronic adenovirus-mediated gene transfer of CTLA4Ig gene and CD40Ig gene result in indefinite survival of islet xenograft. Transpl Proc 35:3165–3166
  7. Konigova R, Matouskova E, Broz L (2000) Burn wound coverage and burn wound closure. Acta Chir Plast 42:64–68
  8. Lari AR, Gang RK (2001) Expansion technique for skin grafts (Meek technique) in the treatment of severely burned patients. Burns 27:61–66
  9. Lenschow DJ, Walunas TL, Bluestone JA (1996) CD28/B7 system of T cell co-stimulation. Annu Rev Immunol 14:233–258
  10. Linsly PS, Wallence PM, Iohnson J (1992) Immuno suppression in vivo by a soluble form of the CTLA4 T cell activation molecule. Science 257:792–797
  11. Lois C, Hong EJ, Pease S, Brown EJ, Baltimore D (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295:868–872
  12. Lui VCH, Tam PKH, Leung MYK et al (2003) Mammary gland-specific secretion of biologically active immunosuppressive agent cytoxic-T-lymphocyte antigen 4 human immunoglobulin fusion protein (CTLA4Ig) in milk by transgenesis. J Immunol Method 277:171–183
  13. Luo GX, Wu J, Chen XW et al (2005) CTLA4Ig introduced by adenoviral vector locally to prolong the survival of xenogeneic skin grafts on rat burn wounds. J Trauma 59:1209–1215
  14. Myoshi H, Blomer U, Takahashi M, Gage FH, Verma IM (1998) Development of a self-inactivating lentivirus vector. J Virol 72:8150–8157
  15. Orgill DP (2009) Excision and skin grafting of thermal burns. New Engl J Med 360:893–901
  16. Pfeifer A (2004) Lentiviral transgenesis. Transgenic Res 13:513–522
  17. Pfeifer A, Ikawa M, Dayn Y, Verma IM (2002) Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc Natl Acad Sci USA 99:2140–2145
  18. Phelps CJ, Ball SF, Vaught TD, Vance AM, Mendicino M, Monahan JA, Walters AH, Wells KD, Dandro AS, Ramsoondar JJ, Cooper DK, Ayares DL (2009) Production and characterization of transgenic pigs expressing porcine CTLA4Ig. Xenotransplantation 16:477–485
  19. Stell D, Marshall H, Bradley JA, Bolton EM (2003) CTLA4-Ig abrogates the anti-globulin response and prolongs cardiac allograft survival after anti-CD2 treatment. Transpl Immunol 12(1):1–7
  20. Tiscornia G, Singer O, Verma IM (2006) Production and purification of lentiviral vectors. Nature Protocls 1:241–245
  21. Vandeput J, Nelissen M, Tanner JC, Boswick J (1995) A review of skin meshers. Burns 21:364–370
  22. Wang YF, Xu AG, Hua YB, Wu WX (2004) Effect of local CTLA4Ig gene transfection on acute rejection of small bowel allografts in rats. World J Gastroenterol 10(6):885–888
  23. Wang Y, Wang FC, Wei H, Ni Y, Wu J, Gao X (2005) Establishement of transgenic mouse line skin-specifically expressing hCTLA4Ig. J Genet Genomes 32:916–922
  24. Wang Y, Ni Y, Wei H, Wang FC, Ge L-P, Gao X (2006) Stable skin-specific overexpression of human CTLA4Ig in transgenic mice through seven generations. Acta Biochim Biophys Sin 38:171–178
  25. Wang YB, Ogawa Y, Kakudo N, Kusumoto K (2007) Survival and wound contraction of full-thickness skin grafts are associated with the degree of tissue edema of the graft bed in immediate excision and early wound excision and grafting in a rabbit model. J Burn Care Res 28:182–186
  26. Wang Y, Wei H, Yong N, Ge LP, Liu Q, Mao XL, Zhao YJ, Wu J (2008) Transgenic expression of cytoxic T lymphocyte associated antigen 4-immunoglobulin prolongs xenogeneic skin graft survival without extensive immunosuppression in rat burn wounds. J Trauma 65:154–162
  27. Wang Y, Song YT, Liu Q, Liu CE, Wang LL, Liu Y, Zhou XY, Wu J, Wei H (2010) Quantitative analysis of lentiviral transgene expression in mice over seven generations. Transgenic Res 19:775–784
  28. Weiner J, Yamada K, Ishikawa Y et al (2010) Prolonged survivial of GalT-KO swine skin on baboons. Xenotransplantation 17:147–152
  29. Whitelaw CBA, Radcliffe PA, Ritchie WA et al (2004) Efficient generation of transgenic pigs using equine anaemia virus (EIAV) derived vector. FEBS Lett 571:233–236
• 作者单位：1. Department of Laboratory Animal Science, College of Basic Medical Sciences, Third Military Medical University, Gao Tan-yan Street, Shapingba District, Chongqing, 400038 China
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Molecular Medicine
  Plant Genetics and Genomics
  Animal Genetics and Genomics
  Plant Sciences
  Human Genetics
  
• 出版者：Springer Netherlands
• ISSN：1573-9368

文摘

Xenogeneic skin, especially porcine skin, has already been used to cover large wounds in clinic practice of wound care. Our previous data showed that transgenic expression of human cytoxic T-lymphocyte associated antigen4-immunoglobulin (hCTLA4Ig) in murine skin graft remarkably prolonged its survival in xenogeneic burn wounds without extensive immunosuppression in recipients, suggesting that transgenic hCTLA4Ig expression in skin graft may be an effective and safe method to prolong its survival in xenogeneic wounds for coverage. Lentiviral transgenesis provides an extremely efficient and cost-effective method to produce transgenic animals. However, tissue-targeted transgenic expression of biologically functional protein by lentiviral transgenesis is rarely reported. In this work, a recombinant lentiviral vector (LV), named FKCW in this article, was constructed by inserting a skin-specific hCTLA4Ig expression cassette consisting of keratin 14 (K14) promoter, hCTLA4Ig coding sequence and an intronic fragment. Its efficacy for transgenesis and skin-specific expression of bio-active hCTLA4Ig protein was tested using mice as models. The LV FKCW was readily to be packaged and concentrated to high titres (1.287–6.254 × 109 TU/ml) by conventional lentivirus package system. Using eggs collected from only five mated females having been subjected to conventional super-ovulation treatment, 8 hCTLA4Ig transgenic founder mice were generated with the concentrated FKCW vector, and transgenic founder per injected and transferred egg was 6.3%, which was nearly 9-fold higher than that for DNA micro-injection with a similar transgene construct in our previous work. The lentiviral transgenic hCTLA4Ig exhibited strictly skin-specific expression at a level comparable to or even slightly higher than that of transgenic hCTLA4Ig delivered by micro-injection in a similar cassette. Lentiviral transgenic hCTLA4Ig protein remarkably suppressed human lymphocyte proliferation in vitro to a degree comparable to that of commercially purchased purified hCTLA4Ig protein with defined activity at similar concentrations. Besides, lentiviral hCTLA4Ig transgenic mouse skin grafted into rat burn wounds exhibited remarkably extended survival compared to wild-type skin of the same strain (13.8 ± 3.8 vs. 6.8 ± 3.0 days), indicating that lentiviral transgenic hCTLA4Ig did inhibit immune rejection against xenogeneic skin graft in vivo. These results laid down the foundation to further efficiently generate transgenic pigs skin-specifically expressing bio-active hCTLA4Ig by lentiviral transgenesis, and provided a demonstration that transgenic animals with tissue-targeted expression of biologically functional protein can be efficiently produced using LV.