A Human Whole Blood Model for Screening Potential Vesicant Antagonists
Abstract
A model employing low-cost, convenient, readily available tissue that consistently expresses a reproducible level of injury is required to screen for potential vesicant antagonists. We have developed a human whole blood, vesicant vapor exposure model. Blood obtained by venipuncture was diluted 40% with RPMI 1640 medium (Gibco/BRL, Grand Island, NY) and 5 ml aliquots dispensed into Costar trays (Allegience Health Care, Lee, MA). Half the samples were exposed for 12 min to 2-chloroethyl ethyl sulfide (CEES) vapor (1.5 mg/L/min), while the other half received carrier gas alone. Following such exposure, samples were incubated at 37 deg C, 5% CO2 for a total of 24 hr. Red blood cells were then lysed (Easy-Lyse(Trademark), Leinco Technologies, St. Louis, MO), and white blood cell viability was measured colorimetrically (ProCheck(Trademark) Cell Viability Assay, Intergen Co., Purchase, NY). CEES exposure decreased viability relative to controls (controls = 100% viability). The viabilities for six CEES exposure trials (2 subjects, 3 times each) ranged from 44.3%-58.2%, with group means (n = 3, by subject) of 52.6% and 48.7%, respectively. These group viabilities were not different from each other, but were significantly (p < .05) depressed compared to controls. This model induced a significant and reproducible level of CEES injury. It appears suitable for the rapid in vitro screening of combinations of potential vesicant antagonists, since the model can accommodate any buffer-soluble, antagonist combination up to a 40% blood displacement.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 2002
- Accession Number
- ADA405432
Entities
People
- David A. Dubose
- Michael D. Blaha
Organizations
- United States Army Medical Research and Development Command