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Development of a modified glucan-specific limulus amebocyte lysate method which correlates murine pulmonary inflammation induced by floor dust collected from a water-damaged building.

Authors
Young-S; Cox-Ganser-JM; Shogren-ES; Wolfarth-MG; Li-S; Antonini-JM; Castranova-V; Park-J
Source
Toxicologist 2010 Mar; 114(1):157
NIOSHTIC No.
20036616
Abstract
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 ~20 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.
Keywords
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
Publication Date
20100301
Document Type
Abstract
Fiscal Year
2010
NTIS Accession No.
NTIS Price
Issue of Publication
1
ISSN
1096-6080
NIOSH Division
HELD
Priority Area
Manufacturing
Source Name
The Toxicologist. Society of Toxicology 49th Annual Meeting and ToxExpo, March 7-11, 2010, Salt Lake City, Utah
State
WV; UT
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