Stable Skin‐specific Overexpression of Human CTLA4‐Ig in Transgenic Mice through Seven Generations

作者：Yong WANG; Yong NI; Hong WEI; Feng‐Chao WANG; Liang‐Peng GE; Xiang GAO

来源：Acta Biochimica et Biophysica Sinica, 2006, 38(3): 171-178.

DOI：10.1111/j.1745-7270.2006.00144.x

• cytotoxic T lymphocyte antigen 4‐Ig
• transgenic mice
• skin
• K14 promoter

Abstract\n\t\t\t\t\t Skin graft rejection is a typical cellular immune response, mainly mediated by T cells. Cytotoxic T lymphocyte associated antigen 4‐immunoglobin (CTLA4‐Ig) extends graft survival by blocking the T cell co‐stimulation pathway and inhibiting T cell activation. To investigate the efficacy of CTLA4‐Ig in prolonging skin graft survival, human CTLA4‐Ig (hCTLA4‐Ig) was engineered to overexpress in mouse skin by transgenesis using the K14 promoter. Reverse transcription‐polymerase chain reaction (
RT‐PCR) and Western blot assay indicated that the expression of CTLA4‐Ig remained skin‐specific and relatively constant compared to the internal control protein, AKT, through seven generations. The presence and concentration of the hCTLA4‐Ig protein in transgenic mouse sera was determined by enzyme‐linked immunosorbent assay (ELISA), and the results indicated that the serum CTLA4‐Ig concentration also remained constant through generations. Survival of transgenic mouse skins grafted onto rat wounds was remarkably prolonged compared to that of wild‐type skins from the same mouse strain, and remained comparable among all seven generations. This suggested that the bioactive hCTLA4‐Ig protein was stably expressed in transgenical mice through at least seven generations, which was consistent with the stable skin‐specific CTLA4‐Ig expression. The results demonstrated that the transgenic expression of hCTLA4‐Ig in skin driven by the K14 promoter remained constant through generations, and a transgenic line can be established to provide transgenic skin with extended survival reproducibly. Edited by 
Zhu‐Chuan ZHANG

• 单位
  西南大学; 南京大学