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Letters to the Editor
Sex-dependent membranopathy in stored human red blood cells
Over the past years, concern has been raised over the impact of red blood cell (RBC) alterations, related to pro- longed hypothermic storage, on patient safety. Multiple reviews and works focus on presentation of a full list of changes characteristic for RBC storage lesions including alterations in RBC metabolomics, cytosol structure and membrane organization.1–6 We recently showed that a decrease in RBC deformability was observed as a relative-
ly late consequence of packed RBC (pRBC) storage, while the analysis of vesiculation observed on the RBC mem- branes in nanoscale with the application of atomic force microscopy (AFM) supported by analysis of biochemical parameters allowed for the detection of early changes.7 However, these results, as well as those of other groups were mainly obtained from pRBC of male donors, or the sex of donors was not specified.5,8 Recently, it has also been suggested that RBC from females are less prone to storage lesion and age slower than male erythrocytes9,10 but these reports were mainly focused on metabolic and functional RBC analysis.
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Figure 1. Storage time-dependent changes in biochemical, morphological and functional red blood cell parameters. Biochemical parameters: (A) cholesterol, (B) triglycerides, (C) lactate dehydrogenase (LDH), (D) free iron of human packed red blood cells (pRBC), (E) glucose, (F) lactate; chosen protein expression: (G) CD47 and (H) phosphatidylserine (PS); (I) pRBC deformability; the blood count including: (J) mean corpuscular volume (MCV), (K) hematocrit (HCT), (L) mean cor- puscular hemoglobin concentration (MCHC). The blood was withdrawn from men (n=12) and women (n=12). Results (A to F) were obtained from pRBC super- natant with ABX Pentra 400 (Horiba Medical, Japan) are given as mean ± standard deviation (SD), data distribution is presented as box plots (mean value, mean ± SD, min-max whiskers). Results (G to L) presented as box plots (median, Q1, Q3, interquartile range, min-max whiskers). Q1, Q3 indicate 25th and 75th per- centiles, respectively. Results (G and H) were obtained using a mixture of mouse anti-human antibodies carrying fluorescent markers: CD45 – APC-Cy7 (1:200, BD cat. no. 348815), CD71 – APC (1:200, BD cat. no. 551374), CD47 – PE (1:200, BD cat. no. 556046) and annexin V – FITC (1:200, BD cat. no. 556547). Cellular analysis was performed with LSRII flow cytometer (BD Biosciences). RBC deformability was studied at 37°C as elongation index at shear rates of 20 Pa with the use of ektacytometer RheoScanAnD 300 (RheoMeditech, Seoul, Korea) while parameters (J and L) with hematology analyzer Abc Vet (Horiba Medical, France). Data (A to L) normality was assessed using Shapiro-Wilk test. The significance of the differences between men and women in each week was evaluated by One-Way ANOVA with Tukey’s test. If data distribution was not normal, non-parametric Kruskal-Wallis test with the Dunn’s post hoc test if appropriate; *P<0.05, **P<0.01, ***P<0.001 ****P<0.0001 indicates significant difference between men and women in each week.
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