Modeling transfusion reactions and predicting in vivo cell survival with kodecytes

The availability of suitable animal models is a limitation in research on transfusion reactions. KODE technology allows for the artificial attachment of incompatible blood group antigens, plus visualization and recovery constructs onto red blood cells (RBCs), making them potentially suitable to study both transfusion reactions and determine in vivo cell survival.

Function-spacer-lipid (FSL) constructs representing blood group A antigen (FSL-A) and biotin (FSL-biotin) together with FSL-GB3 as a benign antigen were used to create a range of murine kodecytes. Compatible and incompatible kodecytes were transfused into naive or anti-A hyperimmune mice. FSL-biotin constructs simultaneously included in the same RBC were visualized with avidin Alexa Fluor 488 and fluorescence microscopy to determine circulating cell survival or to demonstrate recovery of kodecytes.

Using fluorescence microscopy of blood films, biotin, and GB3+biotin kodecyte transfusions, all showed similar survival and were present in the circulation at more than 72 hours in naive and anti-A-immunized mice. A+ biotin kodecytes were also present in the circulation at more than 72 hours in naive mice but were mostly cleared within 6 minutes in anti-A-immunized mice. Avidin agarose beads in gel cards were used to demonstrate recovery of A+ biotin kodecytes from blood samples.

KODE technology not only enables the creation of artificial transfusion reactions in animal models, but also has the potential to be used clinically in man to determine 24-hour cell survival. The ability to recover the kodecytes for further analysis has valuable research and diagnostic potential.