Fig 1: Immune responses and biochemical analysis of organ functions following immunostimulation in a rabbit model.(A) Timeline of prime, boost, and analysis steps of the immunostimulation. (B) Antibody titers in rabbits after receiving immunostimulation with native human RhD-positive RBCs, engineered human RhD-positive RBCs, and human RhD-negative RBCs at different time points. (C) Antibody titers in rabbits during the 5th week of immunostimulation after injection in the four groups. (D) Biochemical analysis of liver function during the 5th week of immunostimulation after injection in the four groups. ALT (U/l), AST (U/l), ALP (U/l), TBIL (μM), DBILALT (μM). (E) Biochemical analysis of kidney function during the 5th week in the four groups after immunostimulation. CREA (μM), BUN (mM), UA (μM).
Fig 2: Rational surface crosslinking framework for the construction of universal RBCs.(A) Schematic illustration of the procedure for RBC surface engineering. (B) Synthetic scheme for the anchor molecule conjugate (BAM-HRP). (C) Mechanism of the formation of PSA-tyramine conjugate and enzymatically crosslinked framework. SEM images of (D) human native RBCs and (G) surface-engineered human RBCs in the presence of BAM-HRP solution, PSA-tyramine (10 mg/liter), and H2O2 solution. TEM images of (E) human native RBCs and (H) surface-engineered human RBCs. Confocal images of (F) native RBCs and (I) engineered RBCs. (J) Optical images of human RhD-positive RBCs in the corresponding anti-typing sera. (K) BAM-tyramine-PSA engineered RhD-positive RBCs were mixed with equal concentrations of their anti-typing sera. (L) Flow cytometry analysis of native RBCs (left) and surface-engineered human RBCs (right) with fluorescein isothiocyanate–labeled anti-RhD antibodies.
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