Evaluation of the sensitivity of red blood cell markers to detect photodynamic membrane damage.
BACKGROUND
Photodynamic dyes have been used to investigate pathogen reduction methods in RBC suspensions, but treatment with sensitizers and light can produce unwanted membrane damage during routine blood bank storage. This study compares the relative sensitivity of three assays for detecting RBC membrane damage.


METHODS
RBCs were treated with dimethylmethylene blue and red light-emitting diode light under four conditions differing in photodynamic stringency and subsequently stored under refrigerated conditions for up to 42 days. Hemolysis, potassium release, and the apoptosis marker annexin V binding were assayed immediately after phototreatment and following storage.


RESULTS
In terms of increasing sensitivity for detecting photodynamic RBC membrane damage was enhanced ion leakage >> hemolysis > annexin V binding.


CONCLUSIONS
Although very stringent photodynamic treatment conditions generate annexin V-positive RBCs, the assay will not detect more subtle membrane damage that is readily detected by other well-established methods. In addition, many RBCs destined for photo-induced hemolysis later in storage are annexin V-negative throughout the storage period, suggesting that the two measures are not directly related.
Wagner
Stephen J
SJ
Blood and Cell Therapy Development, American Red Cross, Biomedical Research and Development, Rockville, Maryland 20855, USA. wagners@usa.redcross.org
Skripchenko
Andrey
A
Thompson-Montgomery
Dedeene
D
Awatefe
Helen
H
D017304:Annexin A5 / Q000378:metabolism*; D015415:Biomarkers; D004910:Erythrocyte Membrane / Q000187:drug effects*; D004912:Erythrocytes / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D010778:Photochemotherapy; D011188:Potassium / Q000378:metabolism*; D012680:Sensitivity and Specificity
D017304:Annexin A5; D015415:Biomarkers; D011188:Potassium