Red blood cell distribution width (RDW) is a biomarker associated with a variety of clinical outcomes. While anemia and subclinical inflammation have been posed as underlying pathophysiology, it is unclear what mechanisms underlie these assocations. Hence, we aimed to unravel the mechanisms in silico using a large clinical dataset and validate our findings in vitro. We retrieved complete blood counts (CBC) from 1,403,663 measurements from the Utrecht Patient Oriented Database, to model RDW using gradient boosting regression. We performed (sex-stratified) analyses in patients with anemia, patients younger/older than 50 and validation across platforms and care settings. We then validated our hypothesis regarding oxidative stress using an in vitro approach. Only percentage microcytic (pMIC) and macrocytic (pMAC) erythrocytes and mean corpuscular volume were most important in modelling RDW (RMSE = 0.40, R2 = 0.96). Subgroup analyses and validation confirmed our findings. In vitro induction of oxidative stress underscored our results, namely increased RDW and decreased erythrocyte volume, yet no vesiculation was observed. We found that erythrocyte size, especially pMIC, is most informative in predicting RDW, but no role for anemia or inflammation. Oxidative stress affecting the size of the erythrocytes may play a role in the association between RDW and clinical outcomes.

Neutrophils have an important role in the immune response. These cells can be subjected to an impaired function and a shift in population depending on disease states. In sepsis, this shift is recognized and flagged by automated hematology analyzers, including the presence of band neutrophils, while these cells, although present, appear not to be detected in trauma patients. To better understand this suspected error in flagging, we set out to distinguish neutrophil populations of these two patient groups and compared these with controls. Different data-driven methods were used compared to standard algorithms used by the software of the analyzers. Using K-means clustering, we extracted neutrophils from raw hematology analyzer datafiles, and compared characteristics of these clusters between the patient groups. We observed an increased neutrophil size for both sepsis and trauma patients, but trauma patients had a smaller increase. Trauma patients also had a high proportion of cells with relatively high nuclear segmentation, which is contradictory with the presence of band neutrophils. This, in combination with the smaller size increase, might explain the inability to flag band neutrophils in trauma.

Red blood cell distribution width (RDW) is a biomarker associated with a variety of clinical outcomes. While anemia and subclinical inflammation have been posed as underlying pathophysiology, it is unclear what mechanisms underlie these associations.