Evaluating ex vivo Toll-like receptor agonist response as a potential biomarker in equine gastric ulcer syndrome.

LPS-stimulated PBMC IL-1β release does not reliably detect or grade EGUS in horses.

This prospective cohort study investigated whether interleukin-1 beta (IL-1β) release from peripheral blood mononuclear cells (PBMCs) following ex vivo Toll-like receptor (TLR) stimulation could serve as an objective biomarker for Equine Gastric Ulcer Syndrome (EGUS) presence and severity. Seventy-seven horses underwent venous blood collection; PBMCs were isolated and stimulated with TLR2 (Pam3CSK4) and TLR4 (lipopolysaccharide, LPS) agonists to quantify IL-1β production. TLR2 stimulation failed entirely, with all samples below the lower limit of quantification. After excluding horses with lameness or systemic inflammation, EGUS prevalence was 92.2% in the cohort. Only 25% of horses demonstrated LPS-induced IL-1β responses. Statistical analyses—including Fisher's exact test, Cochrane-Armitage trend testing, and ROC curve analysis—revealed no significant association between IL-1β response and EGUS status (OR 0.47; p=0.6). Diagnostic performance was poor across all EGUS categories: overall EGUS (AUROC 0.39), Equine Squamous Gastric Disease (AUROC 0.30), and Equine Glandular Gastric Disease (AUROC 0.56). The authors conclude that this LPS-induced PBMC IL-1β release protocol does not differentiate EGUS-positive from EGUS-negative horses. The search for reliable, objective biomarkers of EGUS-associated pain and disease severity continues, with gastroscopy remaining the definitive diagnostic tool. Future biomarker research may need to explore alternative immunological pathways or novel analytes.

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