Red cell supernatant effects on endothelial cell function and innate immune activation is influenced by donor age and sex

Abstract

Background and objectives Blood donor characteristics significantly affect the pre-storage and post-storage quality of red cell concentrates (RCCs). This study investigated the impact of donor age and sex on RCC characteristics and the effect of RCC supernatants on the vascular endothelium and monocytes.Materials and methods RCC units selected from four donor groups based on donor age and sex (male ≤ 30-years old (yo), male ≥ 60-yo, female ≤ 30-yo and female ≥ 60-yo) were stored at 1-6°C and tested on day 7, 21 and 42. RCC supernatants were analysed for the number, concentration and size of extracellular vesicles (EVs). Human vein endothelial cells (HUVECs) monolayers were incubated with RCC supernatants and assessed for expression of vascular cell adhesion molecule-1 (VCAM-1) and E-selectin by flow cytometry as well as endothelial permeability using a trans-epithelial-electrical-resistance assay. Monocytes and HUVECs were incubated with RCC supernatants and supernatants assayed for cytokines/chemokines. ResultsRCCs from female donors were found to have lower unit volume, haemoglobin content and haematocrit compared to RCCs from male donors (P < 0Á05). RCC supernatants from female ≥ 60-yo had a higher concentration of large EVs (≥200 nm) compared to both male ≤ 30-yo (P = 0Á039) and male ≥ 60-yo supernatants (P = 0Á019). Treatment with RCC supernatants from male ≤ 30-yo donors induced lower expression of VCAM-1, lower cytokine/chemokine release and showed the least effect on endothelial permeability.Conclusion Donor characteristics affect RCC quality parameters and influence the endothelial and immune modulation potential of RCC supernatants in vitro.

The Effect of Sex-Mismatched Red Blood Cell Transfusion on Endothelial Cell Activation in Critically Ill Patients

“…It can be hypothesized that in the critically ill patients the presence of an activated endothelium interacts with RBCs, which is in line with previous studies showing that RBC transfusion is associated with changes in ICAM-1 and syndecan-1 concentrations [19]. A recent study has also shown that blood donor age and sex can impact secretion of endothelial markers after incubating blood product supernatant with endothelial cells [20]. In terms of potential mediators of endothelial activation following RBC transfusion, an increase in levels of circulating inflammatory cytokines, such as interleukin-6 (IL-6) and necrosis factor-alpha (TNFα) [6,21], have been suggested to promote endothelial activation [22].…”

Platelet vesicles are potent inflammatory mediators in red blood cell products and washing reduces the inflammatory phenotype

“…The MTM used in this study was adapted from Branch et al and performed as previously described . One mL (2.0 × 10/mL) of the prepared PBMCs suspension was incubated for one hour at 37°C with 5% CO 2 to allow the monocytes to adhere to each of the coverslips that were placed in 35‐mm culture dishes.…”

“…Cryopreserved HUVECs (C2519A, Lonza) were purchased as pooled primary cells (≥500,000 cells/cryovial) and cultured . HUVECs at a concentration of 0.2 × 10 6 cells/mL were seeded into 12‐well flat‐bottom culture plates.…”

Red Blood Cell Donor Sex Associated Effects on Morbidity and Mortality in the Extremely Preterm Newborn

“…This is a potential predictive marker for transfusion associated adverse outcomes and has recently been shown to be increased with sex mismatched RBC transfusion exposure [ 69 ]. In vitro data supports the release of markers of endothelial activation following the incubation of endothelial cells with blood product supernatant [ 70 , 71 ]. Clinical data reports increased ICAM-1, a marker of endothelial activation, and syndecan-1, which is released with the loss of endothelial glycocalyx integrity, following RBC transfusion [ 72 ].…”