Human HLA-Ev (147) Expression in Transgenic Animals.

Area of Science:

• Immunology
• Genetics
• Transplantation Biology

Background:

• Previous work established S147C for HLA-E substitution in xenotransplantation.
• This study focuses on codon exchange for HLA-Ev (147) to enhance its utility.

Purpose of the Study:

• To establish a functional, codon-optimized HLA-Ev (147) gene for xenotransplantation.
• To generate and characterize transgenic mice expressing the modified gene.

Main Methods:

• Constructed a new plasmid with codon-exchanged HLA-Ev (147), IRES, and human beta 2-microglobulin in a pCXN2 vector.
• Assessed natural killer cell and macrophage cytotoxicity in pig endothelial cells transfected with the construct.
• Produced transgenic mice via microinjection and confirmed gene expression in various organs.

Main Results:

• Successfully transfected pig endothelial cells, confirming molecule expression.
• Demonstrated that the expressed molecules effectively regulated natural killer cell and macrophage cytotoxicity.
• Generated multiple lines of transgenic mice with confirmed mRNA expression of HLA-Ev (147) across organs, notably high in the heart and lungs.

Conclusions:

• Codon optimization of HLA-Ev (147) for mammalian systems is crucial for developing effective transgenic animals for xenotransplantation.
• The developed transgenic model provides a valuable tool for further xenotransplantation research.