Giant Multinucleated Cells Are Associated with Mastocytic Inflammatory Signature Equine Asthma

SIMPLE SUMMARY: Lung inflammation is commonly assessed in asthmatic horses by bronchoalveolar lavage fluid (BALF) cytology. Among the cell types commonly found in equine BALF samples, macrophages are the most abundant. The clinical significance of the abnormal cytological appearance of macrophages in samples from diseased horses is largely disregarded. The present work focuses on cytological alterations observed in macrophages during a chronic inflammatory disease such as equine asthma. Our data, although limited in number, support macrophage fusion (resulting in multinucleated giant cells) as a mechanism of interest in the classification of equine asthma phenotypes, as it was significantly associated with the inflammatory signature and chronicity of the disease. ABSTRACT: Equine asthma is currently diagnosed by the presence of increased neutrophil (>5%), mast cell (>2%), and/or eosinophil (>1%) differential cell count. Macrophages are normal resident cells within the alveoli. Their presence in BALF is considered normal, but the clinical implication of the presence of activated or fused macrophages (giant multinucleated cells, GMC) is currently overlooked. We aimed to assess the prevalence, cytological determinants, and clinical significance of increased GMC counts in BALF of 34 asthmatic horses compared to 10 controls. Counts were performed on 15 randomly selected high magnification fields per cytospin slide (40×), and expressed as GMC:single macrophage (GMC:M) ratio. Regression models were used for statistical analysis. GMC was frequently observed in both asthmatic and control horses, with an increased prevalence of equine asthma (p = 0.01). GMC:M ratio was significantly higher in severe vs. mild to moderate equine asthmatic and control horses. In asthmatic horses, an increased GMC:M ratio was significantly associated with BALF mastocytosis (p = 0.01), once adjusting for age and the presence and severity of clinical signs of the horses. Tachypnea was the only clinical sign that tended to be positively associated with GMC:M ratio after adjustment (p = 0.08). In conclusion, our data suggest that a relationship might exist between molecular mechanisms regulating GMC formation and mast cell recruitment in the equine lung. The same mechanisms could lead to tachypnea even in the absence of respiratory effort at rest. We suggest including GMC count in the basic cytological assessment of BALF samples to gain more insights into their role in equine asthma.