1C3-beta-glucan, a major cell wall component of fungi, induces pulmonary inflammation. There is inconsistency in correlation between the levels of glucan measured by current detection methods and the respiratory inflammation observed in people or laboratory animals exposed to fungi. There is a need for a method that better assesses the inflammatory potential of 1C3-beta-glucans in environmental samples. We used the glucan-specific (G-specific) Limulus Amebocyte Lysate (LAL) method after extraction with dimethyl sulfoxide (DMSO), sodium hydroxide (NaOH), or water to analyze the glucan content in floor dusts from a water-damaged building. C3HeB/FeJ mice, an endotoxin-sensitive strain, were treated with different dusts (2.5 mg/kg of body weight), or saline (vehicle control) by pharyngeal aspiration. At 18 hr after aspiration, bronchoalveolar lavage (BAL) was performed, and lung inflammation and injury were assessed by measuring: (1) neutrophil (PMN) infiltration, (2) inflammatory cytokines (IL-6, IL-10, MCP-1, IFN-y, TNF-alpha, and IL12- p70) levels and (3) albumin and lactate dehydrogenase in recovered BAL fluid. Both DMSO and NaOH extaction increased the detection of glucan by approximately20 fold compared to water extraction. Only the DMSO extraction method showed a statistically significant correlation (p<0.05) between 1C3-beta-glucan levels and albumin, total BAL cells, PMNs recovered, TNF-alpha, MCP-1, and IL-6. As expected, significant positive correlations (p<0.05) were also found comparing endotoxin levels and BAL cell numbers, PMNs recovered, IL-6, IFN-y and MCP-1. In conclusion, DMSO extraction for glucan analysis may prove useful in understanding the impact of environmental contamination of glucans on lung disease.
Biological-effects; Cell-biology; Cell-damage; Cell-function; Cellular-reactions; Dust-exposure; Dust-particles; Endotoxins; Environmental-exposure; Environmental-pollution; Exposure-assessment; Exposure-levels; Exposure-methods; Fungal-diseases; Fungal-infections; Fungi; Inhalation-studies; Laboratory-animals; Laboratory-testing; Lung-cells; Lung-disorders; Lung-function; Lung-irritants; Particle-aerodynamics; Particulate-dust; Pulmonary-disorders; Pulmonary-system-disorders; Respiratory-hypersensitivity; Respiratory-infections; Respiratory-irritants; Statistical-analysis; Toxic-effects
The Toxicologist. Society of Toxicology 49th Annual Meeting and ToxExpo, March 7-11, 2010, Salt Lake City, Utah