Red blood cell populations and membrane levels of peroxiredoxin 2 as candidate biomarkers to reveal blood doping.
BACKGROUND
Blood doping represents one main trend in doping strategies. Blood doping refers to the practice of boosting the number of red blood cells (RBCs) in the bloodstream in order to enhance athletic performance, by means of blood transfusions, administration of erythropoiesis-stimulating substances, blood substitutes, natural or artificial altitude facilities, and innovative gene therapies. While detection of recombinant EPO and homologous transfusion is already feasible through electrophoretic, mass spectrometry or flow cytometry-based approaches, no method is currently available to tackle doping strategies relying on autologous transfusions.


METHODS
We exploited an in vitro model of autologous transfusion through a 1:10 dilution of concentrated RBCs after 30 days of storage upon appropriate dilution in freshly withdrawn RBCs from the same donor. Western blot towards membrane Prdx2 and Percoll density gradients were exploited to assess their suitability as biomarkers of transfusion.


RESULTS
Membrane Prdx2 was visible in day 30 samples albeit not in day 0, while it was still visible in the 1:10 dilution of day 30 in day 0 RBCs. Cell gradients also highlighted changes in the profile of the RBC subpopulations upon dilution of stored RBCs in the fresh ones.


CONCLUSIONS
From this preliminary in vitro investigation it emerges that Prdx2 and RBC populations might be further tested as candidate biomarkers of blood doping through autologous transfusion, though it is yet to be assessed whether the kinetics in vivo of Prdx2 exposure in the membrane of transfused RBCs will endow a sufficient time-window to allow reliable anti-doping testing.
Department of Ecological and Biological Sciences, University of Tuscia, Largo dell'Universita snc., Viterbo, Italy.
Cristina
C
Marrocco
Valeria
V
Pallotta
Angelo
A
D'alessandro
Gilda
G
Alves
Lello
L
Zolla
D000328:Adult; D015415:Biomarkers / Q000032:analysis; D004300:Doping in Sports; D004910:Erythrocyte Membrane / Q000737:chemistry*; D004912:Erythrocytes; D005260:Female; D006801:Humans; D008297:Male; D054464:Peroxiredoxins / Q000032:analysis*
D015415:Biomarkers; D054464:Peroxiredoxins