Co-expression of the complement regulatory proteins human DAF and CD59 with an IRES-mediated dicistronic mammalian vector enhances their cell protective effects

  • Authors:
    • Li Xu
    • Huanjie Shao
    • Wenlan Wu
    • Zhijian Cao
    • Zhouzhou Zhao
    • Hui Liu
    • Dahe Jiang
    • Xin Mao
    • Wenxin Li
  • View Affiliations

  • Published online on: September 1, 2005     https://doi.org/10.3892/ijmm.16.3.409
  • Pages: 409-414
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The human complement regulatory proteins (hCRPs) decay accelerating factor (DAF/CD55) and protectin CD59 transfected into non-human cells could confer protection against human complement. The combination of DAF and CD59 would be an effective strategy to help overcome host complement-induced hyperacute rejection in xenotransplantation. We constructed a dicistronic mammalian expression vector pcDNA3-CD59IRESDAF by using the internal ribosomal entry sites (IRES) of the encephalomyocarditis virus (EMCV). RT-PCR, Western blotting and immunofluorescence microscopic analysis demonstrated that the EMCV IRES allowed for efficient co-expression of hCD59 and hDAF on the surface of NIH/3T3 cells transfected stably with pcDNA3-CD59IRESDAF. Human complement-mediated cytolysis assays showed that co-expressed DAF and CD59 proteins could provide more significant protection against complement-mediated cytolysis than either hCD59 or hDAF alone. These results suggest that IRES containing polycistronic vector should improve the efficiency and effectiveness of multi-gene delivery and that the construct pcDNA3-CD59IRESDAF vector has potential therapeutic value for effectively controlling complement activation and for preventing hyperacute rejection in clinical gene therapy.

Related Articles

Journal Cover

September 2005
Volume 16 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Xu L, Shao H, Wu W, Cao Z, Zhao Z, Liu H, Jiang D, Mao X and Li W: Co-expression of the complement regulatory proteins human DAF and CD59 with an IRES-mediated dicistronic mammalian vector enhances their cell protective effects. Int J Mol Med 16: 409-414, 2005.
APA
Xu, L., Shao, H., Wu, W., Cao, Z., Zhao, Z., Liu, H. ... Li, W. (2005). Co-expression of the complement regulatory proteins human DAF and CD59 with an IRES-mediated dicistronic mammalian vector enhances their cell protective effects. International Journal of Molecular Medicine, 16, 409-414. https://doi.org/10.3892/ijmm.16.3.409
MLA
Xu, L., Shao, H., Wu, W., Cao, Z., Zhao, Z., Liu, H., Jiang, D., Mao, X., Li, W."Co-expression of the complement regulatory proteins human DAF and CD59 with an IRES-mediated dicistronic mammalian vector enhances their cell protective effects". International Journal of Molecular Medicine 16.3 (2005): 409-414.
Chicago
Xu, L., Shao, H., Wu, W., Cao, Z., Zhao, Z., Liu, H., Jiang, D., Mao, X., Li, W."Co-expression of the complement regulatory proteins human DAF and CD59 with an IRES-mediated dicistronic mammalian vector enhances their cell protective effects". International Journal of Molecular Medicine 16, no. 3 (2005): 409-414. https://doi.org/10.3892/ijmm.16.3.409