Discriminating complement-mediated acute transfusion reaction for type O+ red blood cells transfused into a B+ recipient with the complement hemolysis using human erythrocytes (CHUHE) assay

Transfusion. 2016 Jul;56(7):1845-8. doi: 10.1111/trf.13592. Epub 2016 Mar 20.


Background: A patient with B+ sickle cell disease received 3 units of red blood cells (RBCs) from two O+ donors and developed fever and hypotension after the first unit, consistent with an acute transfusion reaction (ATR). Anti-B titers in plasma from each O+ donor were markedly elevated and nondiscriminatory. In order to evaluate the potential for the transfused units to produce complement-mediated hemolysis of B+ RBCs, hemolytic complement testing was performed.

Study design and methods: Plasma from each donor was diluted in veronal buffer and incubated with B+ RBCs, and free hemoglobin was measured by spectrophotometer in the complement hemolysis using human erythrocytes (CHUHE) assay. Peptide inhibitor of complement C1 (PIC1) was used to confirm antibody-initiated complement pathway activation.

Results: A 96-fold difference (p = 0.014) in hemolysis was measured between plasma samples from the two O+ donors using the CHUHE assay. The extremely high degree of hemolysis produced by the one plasma was inhibited by PIC1 in a dose-dependent manner.

Conclusion: These results indicate that hemolytic complement testing with the CHUHE assay can be used to assess the risk of antibody-initiated, complement-mediated hemolysis from a transfusion beyond what can be achieved with antibody titers alone.

MeSH terms

  • ABO Blood-Group System / immunology*
  • Adolescent
  • Anemia, Sickle Cell / therapy
  • Antibodies
  • Blood Group Incompatibility
  • Complement Activation / immunology*
  • Female
  • Hemolysis / immunology*
  • Humans
  • Middle Aged
  • Risk Assessment
  • Transfusion Reaction / immunology*


  • ABO Blood-Group System
  • Antibodies