Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Oct;92(4):1831–1839. doi: 10.1172/JCI116774

Epitope mapping of type VII collagen. Identification of discrete peptide sequences recognized by sera from patients with acquired epidermolysis bullosa.

J C Lapiere 1, D T Woodley 1, M G Parente 1, T Iwasaki 1, K C Wynn 1, A M Christiano 1, J Uitto 1
PMCID: PMC288347  PMID: 7691888

Abstract

Epidermolysis bullosa acquisita (EBA) is an acquired blistering skin disease characterized by the presence of IgG autoantibodies that recognize type VII (anchoring fibril) collagen. In this study, we have mapped the antigenic epitopes within the type VII collagen alpha chain by Western immunoblotting analysis with sera from 19 patients with EBA, using bacterial collagenase- or pepsin-resistant portions of type VII collagen and a panel of 12 recombinant fusion proteins corresponding to approximately 80% of the primary sequence of the alpha 1 (VII) collagen polypeptide. These studies identified four major immunodominant epitopes localized within the amino-terminal, noncollagenous (NC-1) domain. In addition to EBA, sera from three patients with bullous systemic lupus erythematosus (BSLE) were tested. The pattern of epitopes recognized by these sera were similar to those noted with EBA, suggesting that the same epitopes could serve as autoantigens in both blistering conditions. In contrast, sera from healthy controls or from patients with unrelated blistering skin diseases did not react with type VII collagen epitopes. Collectively, the results indicate that the immunodominant epitopes in EBA and BSLE lie within the noncollagenous regions of type VII collagen. The precise role of the circulating autoantibodies in the pathogenesis of these blistering diseases remains to be elucidated. Conceivably, however, such antibodies could disrupt the assembly of type VII collagen into anchoring fibrils and/or interfere with their interactions with other extracellular matrix molecules within the cutaneous basement membrane zone.

Full text

PDF
1831

Images in this article

1833Image
on p.1833
1834Image
on p.1834
1834Image
on p.1834
1834Image
on p.1834
1834Image
on p.1834
1834Image
on p.1834
1834Image
on p.1834
1836Image
on p.1836
1836Image
on p.1836

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anhalt G. J., Kim S. C., Stanley J. R., Korman N. J., Jabs D. A., Kory M., Izumi H., Ratrie H., 3rd, Mutasim D., Ariss-Abdo L. Paraneoplastic pemphigus. An autoimmune mucocutaneous disease associated with neoplasia. N Engl J Med. 1990 Dec 20;323(25):1729–1735. doi: 10.1056/NEJM199012203232503. [DOI] [PubMed] [Google Scholar]
  2. Christiano A. M., Rosenbaum L. M., Chung-Honet L. C., Parente M. G., Woodley D. T., Pan T. C., Zhang R. Z., Chu M. L., Burgeson R. E., Uitto J. The large non-collagenous domain (NC-1) of type VII collagen is amino-terminal and chimeric. Homology to cartilage matrix protein, the type III domains of fibronectin and the A domains of von Willebrand factor. Hum Mol Genet. 1992 Oct;1(7):475–481. doi: 10.1093/hmg/1.7.475. [DOI] [PubMed] [Google Scholar]
  3. Fine J. D., Bauer E. A., Briggaman R. A., Carter D. M., Eady R. A., Esterly N. B., Holbrook K. A., Hurwitz S., Johnson L., Lin A. Revised clinical and laboratory criteria for subtypes of inherited epidermolysis bullosa. A consensus report by the Subcommittee on Diagnosis and Classification of the National Epidermolysis Bullosa Registry. J Am Acad Dermatol. 1991 Jan;24(1):119–135. doi: 10.1016/0190-9622(91)70021-s. [DOI] [PubMed] [Google Scholar]
  4. Fullerton S. H., Woodley D. T., Smoller B. R., Anhalt G. J. Paraneoplastic pemphigus with autoantibody deposition in bronchial epithelium after autologous bone marrow transplantation. JAMA. 1992 Mar 18;267(11):1500–1502. [PubMed] [Google Scholar]
  5. Gammon W. R., Briggaman R. A., Woodley D. T., Heald P. W., Wheeler C. E., Jr Epidermolysis bullosa acquisita--a pemphigoid-like disease. J Am Acad Dermatol. 1984 Nov;11(5 Pt 1):820–832. doi: 10.1016/s0190-9622(84)80459-4. [DOI] [PubMed] [Google Scholar]
  6. Gammon W. R., Inman A. O., 3rd, Wheeler C. E., Jr Differences in complement-dependent chemotactic activity generated by bullous pemphigoid and epidermolysis bullosa acquisita immune complexes: demonstration by leukocytic attachment and organ culture methods. J Invest Dermatol. 1984 Jul;83(1):57–61. doi: 10.1111/1523-1747.ep12261694. [DOI] [PubMed] [Google Scholar]
  7. Gammon W. R., Woodley D. T., Dole K. C., Briggaman R. A. Evidence that anti-basement membrane zone antibodies in bullous eruption of systemic lupus erythematosus recognize epidermolysis bullosa acquisita autoantigen. J Invest Dermatol. 1985 Jun;84(6):472–476. doi: 10.1111/1523-1747.ep12272402. [DOI] [PubMed] [Google Scholar]
  8. Guldner H. H., Netter H. J., Szostecki C., Lakomek H. J., Will H. Epitope mapping with a recombinant human 68-kDa (U1) ribonucleoprotein antigen reveals heterogeneous autoantibody profiles in human autoimmune sera. J Immunol. 1988 Jul 15;141(2):469–475. [PubMed] [Google Scholar]
  9. Kim Y. H., Woodley D. T., Wynn K. C., Giomi W., Bauer E. A. Recessive dystrophic epidermolysis bullosa phenotype is preserved in xenografts using SCID mice: development of an experimental in vivo model. J Invest Dermatol. 1992 Feb;98(2):191–197. doi: 10.1111/1523-1747.ep12555849. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Maul G. G., Jimenez S. A., Riggs E., Ziemnicka-Kotula D. Determination of an epitope of the diffuse systemic sclerosis marker antigen DNA topoisomerase I: sequence similarity with retroviral p30gag protein suggests a possible cause for autoimmunity in systemic sclerosis. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8492–8496. doi: 10.1073/pnas.86.21.8492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nieboer C., Boorsma D. M., Woerdeman M. J., Kalsbeek G. L. Epidermolysis bullosa acquisita. Immunofluorescence, electron microscopic and immunoelectron microscopic studies in four patients. Br J Dermatol. 1980 Apr;102(4):383–392. doi: 10.1111/j.1365-2133.1980.tb06550.x. [DOI] [PubMed] [Google Scholar]
  13. Paller A. S., Queen L. L., Woodley D. T., Gammon W. R., O'Keefe E. J., Briggaman R. A. A mouse monoclonal antibody against a newly discovered basement membrane component, the epidermolysis bullosa acquisita antigen. J Invest Dermatol. 1985 Mar;84(3):215–217. doi: 10.1111/1523-1747.ep12265158. [DOI] [PubMed] [Google Scholar]
  14. Parente M. G., Chung L. C., Ryynänen J., Woodley D. T., Wynn K. C., Bauer E. A., Mattei M. G., Chu M. L., Uitto J. Human type VII collagen: cDNA cloning and chromosomal mapping of the gene. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6931–6935. doi: 10.1073/pnas.88.16.6931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sakai L. Y., Keene D. R., Morris N. P., Burgeson R. E. Type VII collagen is a major structural component of anchoring fibrils. J Cell Biol. 1986 Oct;103(4):1577–1586. doi: 10.1083/jcb.103.4.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sinha A. A., Lopez M. T., McDevitt H. O. Autoimmune diseases: the failure of self tolerance. Science. 1990 Jun 15;248(4961):1380–1388. doi: 10.1126/science.1972595. [DOI] [PubMed] [Google Scholar]
  18. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  19. Tidman M. J., Eady R. A. Evaluation of anchoring fibrils and other components of the dermal-epidermal junction in dystrophic epidermolysis bullosa by a quantitative ultrastructural technique. J Invest Dermatol. 1985 May;84(5):374–377. doi: 10.1111/1523-1747.ep12265460. [DOI] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Uitto J., Bauer E. A., Moshell A. N. Symposium on epidermolysis bullosa: molecular biology and pathology of the cutaneous basement membrane zone. Jefferson Medical College, Philadelphia, Pennsylvania, October 4 and 5, 1991. J Invest Dermatol. 1992 Mar;98(3):391–395. doi: 10.1111/1523-1747.ep12499822. [DOI] [PubMed] [Google Scholar]
  22. Uitto J., Christiano A. M. Molecular genetics of the cutaneous basement membrane zone. Perspectives on epidermolysis bullosa and other blistering skin diseases. J Clin Invest. 1992 Sep;90(3):687–692. doi: 10.1172/JCI115938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Uitto J., Chung-Honet L. C., Christiano A. M. Molecular biology and pathology of type VII collagen. Exp Dermatol. 1992 Jul;1(1):2–11. doi: 10.1111/j.1600-0625.1992.tb00065.x. [DOI] [PubMed] [Google Scholar]
  24. Woodley D. T., Briggaman R. A., O'Keefe E. J., Inman A. O., Queen L. L., Gammon W. R. Identification of the skin basement-membrane autoantigen in epidermolysis bullosa acquisita. N Engl J Med. 1984 Apr 19;310(16):1007–1013. doi: 10.1056/NEJM198404193101602. [DOI] [PubMed] [Google Scholar]
  25. Woodley D. T., Burgeson R. E., Lunstrum G., Bruckner-Tuderman L., Reese M. J., Briggaman R. A. Epidermolysis bullosa acquisita antigen is the globular carboxyl terminus of type VII procollagen. J Clin Invest. 1988 Mar;81(3):683–687. doi: 10.1172/JCI113373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Woodley D. T., O'Keefe E. J., McDonald J. A., Reese M. J., Briggaman R. A., Gammon W. R. Specific affinity between fibronectin and the epidermolysis bullosa acquisita antigen. J Clin Invest. 1987 Jun;79(6):1826–1830. doi: 10.1172/JCI113024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wraith D. C., McDevitt H. O., Steinman L., Acha-Orbea H. T cell recognition as the target for immune intervention in autoimmune disease. Cell. 1989 Jun 2;57(5):709–715. doi: 10.1016/0092-8674(89)90786-1. [DOI] [PubMed] [Google Scholar]
  28. Yoshiike T., Woodley D. T., Briggaman R. A. Epidermolysis bullosa acquisita antigen: relationship between the collagenase-sensitive and -insensitive domains. J Invest Dermatol. 1988 Feb;90(2):127–133. doi: 10.1111/1523-1747.ep12462092. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES