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Related Experiment Videos

Expression of functional decay-accelerating factor (CD55) in transgenic mice protects against human

B J van Denderen1, M J Pearse, M Katerelos

  • 1Immunology Research Centre, St. Vincent's Hospital, Fitzroy, Victoria, Australia.

Transplantation
|February 27, 1996
PubMed
Summary

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Transgenic mice expressing human CD55 demonstrate reduced complement C3 deposition and lysis from human serum. This creates a valuable model for studying complement regulation and xenotransplantation.

Area of Science:

  • Immunology
  • Genetics
  • Transplantation Biology

Background:

  • Complement system plays a crucial role in innate immunity and rejection of foreign tissues.
  • CD55 (DAF) is a key complement regulatory protein that prevents complement-mediated damage.
  • Developing models to study human complement regulation in vivo is essential for advancing xenotransplantation.

Purpose of the Study:

  • To generate and characterize transgenic mice expressing human CD55.
  • To evaluate the functional capacity of transgenic human CD55 in preventing human complement activation.
  • To assess the potential of these mice as a model for xenotransplantation research.

Main Methods:

  • Generation of transgenic mice via microinjection of a human CD55-minigene.

Related Experiment Videos

  • PCR analysis for transgene integration.
  • Flow cytometry for CD55 expression on peripheral blood leukocytes (PBLs).
  • Immunohistochemistry on various tissues.
  • Assessment of C3 deposition on splenocytes using flow cytometry.
  • Evaluation of human serum-mediated lysis using LDH release assay.
  • Langendorff perfusion of hearts with human plasma.

    • Main Results:

    • Transgenic mice successfully expressed human CD55 on PBLs and in various tissues.
    • Transgenic splenocytes showed significantly reduced C3 deposition (approx. 65%) upon incubation with human serum.
    • Human serum-mediated lysis of transgenic cells was significantly reduced.
    • Deposition of human C3c was greatly reduced in perfused transgenic hearts.
    • No increased survival time was observed in perfused hearts.

    Conclusions:

    • Transgenic mice expressing human CD55 exhibit functional protection against human complement-mediated attack.
    • These mice serve as a valuable preclinical model for studying complement regulation and xenotransplantation.
    • Further research is needed to optimize CD55 expression and address other barriers in xenotransplantation.