Rhesus D Antigenic Determinants on Residual Red Blood Cells in Apheresis and Buffy Coat Platelet Concentrates

The level of residual red blood cells (RBCs) in platelet concentrates (PCs) is of interest because of clinical concerns related to alloimmunization to RBC antigens in transfused patients. This work aims at characterizing and quantifying the levels of intact and fragmented RBCs in apheresis (AP-PCs) and buffy coat PCs (BC-PCs) to assess their potential risk for RhD antigen alloimmunization.

After staining with anti-CD41 (platelets) and anti-CD235a (RBCs) antibodies, the size and density of RhD antigen on intact and fragmented RBCs were analyzed by flow cytometry.

Residual RBC counts were 29 ± 22 × 10⁶/unit in AP-PCs and 121 ± 54 × 10⁶/unit in BC-PCs, which correspond to about 3 and 11 µL of RBCs by product, respectively. RhD expression was about 4 times higher on RBC particles in AP-PCs, and these particles contribute to 66 and 75% of the total antigenic load in BC-PCs and AP-PCs, respectively.

Processing methods influence the quantity and nature of contaminating residual RBCs and RBC-derived particles in PCs. The estimation of residual RBCs in these blood products is generally based on measurements of intact RBCs, which might underestimate the risk for alloim-munization in transfused patients. The question of whether these RBC-derived particles can produce an immune response and, thus, should then be taken into consideration for Rh immune prophylactic treatments, remains to be clarified.