Changes in connective tissue macromolecular components of Yucatan mini-pig skin following application of sulphur mustard vapour

Abstract

1 The aim of this study was to determine the nature of the macromolecular alterations in Yucatan mini-pig skin which occur following application of sulphur mustard vapour, with particular reference to laminin and type IV collagen.

2 The immunostaining of transfer blots from skin extracts run on SDS-PAGE gels revealed no evidence of cross-link ing of type IV collagen or laminin. Laminin was, however, found to be partially degraded as determined by the reso lution of 132 and 143 kDa fragments, possibly by the acti vation of proteases, following the application of sulphur mustard to pig skin. Type IV collagen was not subject to this form of degradation in the skin samples exposed to sulphur mustard.

3 Yucatan mini-pig skin was found to develop microblis ters after exposure to sulphur mustard vapour. The immunohistochemical studies of sulphur mustard exposed skin revealed that separation of the epidermis from the dermis was found to occur within the lamina lucida of the subepidermal basement membrane, supporting the con tention that cleavage of laminin networks occurs following mustard challenge. Immunohistochemical staining with anti-type IV collagen antibodies was restricted to the floor of the micro-blister lesions.

4 The results suggest that laminin may be a target for pro tease activation at the dermo-epidermal junction. This may account for the tendency of certain skin models to develop sulphur mustard-induced blistering. The Yucatan mini-pig may be valuable as a model to determine the effi cacy of prophylactic and therapeutic regimes.

Keywords

sulphur mustard pig skin electrophoresis collagen laminin immunoblotting

Get full access to this article

View all access options for this article.

References

Senter TP , Stimpson DH & Charles G. Comparison of two therapeutic regimes using the same topical corticoid for stable psoriasis. Western Journal of Medicine 1983; 139: 657-62.

Fox M. & Scott M. The genetic toxicology of nitrogen and sulphur mustard. Mutation Research 1980; 75: 131-68.

Papirmeister B. , Gross CL , Petrali JP & Hixson C. Pathology produced by sulphur mustard in human skin grafts on athymic nude mice. I. Gross and light microscopic changes. Journal of Toxicology (Occular and Cutaneous Toxicology) 1994; 3: 371-91.

Smulson ME Expression of cloned poly(ADP-Ribose)polymerase domains may alter mustard induced DNA repair. Proceedings of the 1989 Medical Defence Bioscience Review, 15-17 August 1989, US Army Medical Research and Development Command, Maryland, USA, pp. 367-71.

Miskin R. & Reich E. Plasminogen activator : Induction of synthesis by DNA damage . Cell 1980; 19: 217-24.

Singer KH , Hashimoto K. & Lazarus GS Antibody-induced proteinase activation: a proposed mechanism for pemphigus . Springer Seminars in Immunopathology 1981 ; 4: 17.

Sellers A. & Murphy G. Collagenolytic enzymes and their naturally occurring inhibitors. In: International Review of Connective Tissue Research. Hall D.A. & Jackson D.S. (eds.). Academic Press, New York and London, 1981. Volume (9), pages 151-90.

Gross CL , Meier HL , Papirmeister B. , Brinkley FB & Johnson JB Sulphur mustard lowers nicotinamide adenine dinucleotide concentrations in human skin grafted to athymic nude mice. Toxicology and Applied Pharmacology 1985; 81: 85-90.

Dannenberg AM , Rikimaru T. , Yano T. , Nakamura M. , Beck G. , Habicht GS , Rennie LL , Pula PJ & Schofield BH Sulphur mustard (SM) lesions in organ-cultured human skin: Markers of injury and inflammatory mediators. Proceedings of the 1989 Medical Defence Bioscience Review, 15-17 August 1989, US Army Medical Research and Development Command , Maryland, USA, pp. 41-4.

10.

Mitcheltree LW , Mershon MM , Wall HG , Pulliam JD & Manthei JH Microblister formation in vesicant-exposed pig skin. Journal of Toxicology (Occular and Cutaneous Toxicology) 1989; 8: 309-19.

11.

Marlow DD , Mershon MM , Mitcheltree LW , Petrali JP & Jaax GP Sulphur mustard-induced skin injury in hairless guinea-pigs. Journal of Toxicology (Occular and Cutaneous Toxicology) 1990; 9: 179-92.

12.

Bartek MJ , Labudde JA & Maibach HI Skin permeability in vivo: Comparison in rat, rabbit, pig and man . Journal of Investigative Dermatology 1972 ; 58: 114-23.

13.

Tryggvason K. , Hoyhtya M. & Salo T. Proteolytic degradation of extracellular matrix in tumour invasion. Biochima et Biophysica Acta 1987; 907: 191-217.

14.

Pirie A. The action of mustard gas on ox cornea collagen. Biochemical Journal 1947; 41: 185-90.

15.

Miller EJ & Gay S. Collagen : An overview. Methods in Enzymology 1982; 82: 3-32.

16.

Miller EJ & Rhodes RK Preparation and characterisation of the different types of collagen. Methods in Enzymology 1982; 82: 33-64.

17.

Fuji T. & Kuhn K. Isolation and characterisation of pepsin-treated type III collagen from calf skin. Hoppe-Seyler's Zeitschrift fuer Physiologische Chemie 1971; 356: S1793-1801.

18.

Hascall VC & Kimura JH Proteoglycans: Isolation and characterisation. Methods in Enzymology 1982; 82: 769-99.

19.

Bradford M. A practical and sensitive method for the quantitation of microgram quantities of protein utilising the principle of dye binding . Annals of Biochemistry 1976; 72: 248-52.

20.

Timpl R. , Rohde H. , Risteli L. , Ott U. , Robey PG & Martin GR Laminin. Methods in Enzymology 1982; 82: 831-37.

21.

Kleinman HK , Cannon FB & Laurie GW Biological activities of laminin. Journal of Cellular Biochemistry 1985; 27: 317-25.

22.

Bosman FT , Cleutjens J. , Beek C. & Havenith M. Basement membrane heterogeneity. Histochemical Journal 1989; 21: 629-33.

23.

Katz SI The epidermal basement membrane zone: structure, ontogeny and role in disease . Journal of the American Academy of Dermatology 1984; 11: 1025-37.

24.

Abrahamson D. Recent studies on the structure and pathology of basement membranes. Journal of Pathology 1986; 149: 257-78.

25.

Odermatt E. , Furthmayer H. , Timpl R. , Rohde M. , Ott V. & Engel J. Shape, domain structure, and flexibility of laminin. In: New Trends in Basement Membrane Research, Kuehn K , Schoene H-H & Rimpl R (eds), Raven Press, New York, p. 79, 1982.

26.

Foidart JM , Bere EW , Gullino M. , Brown K. , Martin GR , Yaar M. & Katz SI Nature, localisation and function of type IV collagen and laminin in basement membranes. Clinical Research 1979; 27: 446A.

27.

Mershon MM , Mitcheltree LW , Petrali JP , Braue EH & Wade JV Hairless Guinea pig bioassay model for vesicant vapour exposures. Fundamental and Applied Toxicology 1990; 15: 622-30.

28.

Vogt RF , Dannenberg AM , Schofield BH , Hynes NA & Papirmeister B. Pathogenesis of skin lesions caused by sulfur mustard. Fundamental and Applied Toxicology 1984; 4: S71-S83.

29.

Marlow DD , Mersham MM , Mitcheltree LW , Petrali JP & Jaax GC Evaluation of euthymic hairless guinea pigs as an animal model for vesicant injury. Proceedings of the 1989 Medical Defence Bioscience Review, US Army Medical Research and Development Command, 15-17 August 1989, Maryland, USA, pp. 561-8, 1989.

30.

Mol Mae , De Vries R. & Kluivers AW Effects of nicotinamide on biochemical changes and microblistering induced by sulphur mustard in human skin organ cultures. Toxicology and Applied Pharmacology 1991; 107: 439-49.