To investigate indicators of neutrophil activation in the blood of healthy and asthma-affected horses and assess associations between corticosteroid treatment and these variables. 48 horses (14 with severe equine asthma [SEA], 21 with mild to moderate equine asthma [MEA], and 13 healthy controls). In a 3-part retrospective study, hematology analyzer data for horses included in previous studies were reviewed. Neutrophil size, neutrophil light absorbance (NLA), and myeloperoxidase (MPO) index were recorded. Data for each variable were compared among groups for the entire study sample (part 1). Changes in each variable were assessed for one subset of horses (5 SEA-affected and 6 controls) after treatment for 2 weeks with dexamethasone (0.06 mg/kg, PO, q 24 h; part 2) and for another subset (8 SEA-affected horses) after the same treatment and after a 1-week posttreatment washout period (part 3). All 3 variables were significantly greater for the SEA group, compared with the MEA and control groups in part 1. Following dexamethasone treatment, the control- and SEA-group NLA and MPO index significantly decreased and SEA-group neutrophil size significantly decreased in part 2; immediate posttreatment results for SEA-affected horses were similar in part 3, with significantly increased neutrophil size and nonsignificant increases in NLA and MPO index following washout. Results suggested horses with exacerbated SEA have larger neutrophils that contain more MPO, compared with neutrophils of MEA-affected and healthy control horses. The clinical value of these variables for the diagnosis of equine asthma was deemed limited owing to data overlap among groups.