Multiphoton microscopy: an optical approach to understanding and resolving sulfur mustard lesions.
Sulfur mustard (SM; 2,2(')-dichloroethyl sulfide) is a percutaneous alkylating agent first used as a chemical weapon at Ypres, Belgium, in World War I. Despite its well-documented history, the primary lesions effecting dermal-epidermal separation and latent onset of incapacitating blisters remain poorly understood. By immunofluorescent imaging of human epidermal keratinocytes (HEK) and epidermal tissues exposed to SM (400 microM for 5 min), we have amassed unequivocal evidence that SM disrupts adhesion complex molecules, which are also disrupted by epidermolysis bullosa-type blistering diseases of the skin. Images of keratin 14 (K14) in control cells showed tentlike filament networks linking the HEK's basolateral anchoring sites to the dorsal surface of its nuclei. Images from 6-h postexposure profiles revealed early disruption (</=1 h) and progressive collapse of the K14 cytoskeleton. Collapse involved focal erosions, loss of functional asymmetry, and displacement of nuclei beneath a mat of jumbled filaments. In complementary studies, 1-h images showed statistically significant (p<0.01) decreases of 25 to 30% in emissions from labeled alpha(6)beta(4) integrin and laminin 5, plus disruption of their receptor-ligand organization. Results indicate that SM alkylation destabilizes dermal-epidermal attachments and potentiates vesication by disrupting adhesion complex molecules and associated signaling mechanisms required for their maintenance and repair.
Werrlein
Robert J
RJ
U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, Maryland 21010-5400, USA. Robert.Werrlein@amedd.army.mil
Madren-Whalley
Janna S
JS
D015815:Cell Adhesion Molecules / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D002619:Chemical Warfare Agents / Q000633:toxicity; D046508:Culture Techniques; D004817:Epidermis / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D021621:Imaging, Three-Dimensional / Q000379:methods*; D039161:Integrin alpha6beta4 / Q000378:metabolism / Q000648:ultrastructure; D053547:Keratin-14; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D007633:Keratins / Q000378:metabolism / Q000648:ultrastructure; D007797:Laminin / Q000378:metabolism / Q000648:ultrastructure; D008856:Microscopy, Fluorescence / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D009151:Mustard Gas / Q000633:toxicity*; D012867:Skin / Q000293:injuries / Q000378:metabolism / Q000473:pathology; D012871:Skin Diseases / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D014018:Tissue Distribution; D041622:Tomography, Optical / Q000379:methods*
D015815:Cell Adhesion Molecules; D002619:Chemical Warfare Agents; D039161:Integrin alpha6beta4; C507540:KRT14 protein, human; D053547:Keratin-14; D007797:Laminin; D007633:Keratins; D009151:Mustard Gas