Effect of Potential Vesicant Antagonist Drugs on White Blood Cell Metabolic Activity in Human Whole Blood Exposed to 2-Chloroethyl Ethyl Sulfide

摘要

Mustard agent affects several pathways that involve calcium poisoning and protein degradation, but it is not clear how these pathways affect vesicant-induced toxicity. This makes it difficult to develop a good screening test to evaluate chemicals which might be used to antagonize or mitigate mustard's actions in cells. We examined here a screening test that uses live human tissue (human whole blood) and describe how monitoring the tissue after exposure to potential drugs and a vesicant can be used to estimate the overall effect of a drug or combination of drugs on vesicant exposure. Antagonizing cell death pathways of calcium poisoning (zaldaride maleate CGS9343B); C) or protein degradation (Leupeptin; L) does not satisfactorily resolve vesicant-induced toxicity. However, antagonizing pathways simultaneously might be successful. Moreover, when used with an oxidizing agent (copper-based polyoxometalate; P) that may reduce vesicant toxicity, the effectiveness of these antagonists might be improved. This approach was evaluated using a simple human whole blood (HWB) in vitro system. Prior treatment (30 min) of heparinized (40 U/mL) HWB (n=9) without (buffer vehicle) or with C (13.6 microgram/mL), L (0.5 mg/mL) or P (5.0 microgram/mL) singly (N=3) or in combination (N=4) was tested for white blood cell metabolic activity (WBCMA; Intergen ProCheck R)) post (24 h of in vitro culture) carrier (air) gas or vesicant (2-chloroethyl ethyl sulfide; CEES; 18mg at 1.5 mL/min) exposure. With carrier gas, L and/or C, significantly enhanced the measured WBCMA, while P sustained WBCMA, when compared to HWB without drug(s). This salutary, in vitro effect explained why L significantly improved or C, P, C+L, C+P or L+P sustained WBCMA with CEES, when compared to HWB without drug(s). However, when drug(s) was present in both the CEES and carrier gas conditions, the WBCMA was significantly reduced by CEES exposure.