Skip to main content
SpringerLink
Log in

Application of Direct Immunofluorescence for Skin and Mucosal Biopsies: A Practical Review

  • Chapter
  • First Online:
Book cover Handbook of Practical Immunohistochemistry

Abstract

This is a practical overview of the use of the application of direct immunofluorescence, written in telegraphic style, based on the author’s personal experience, and supplemented by a recommended reference reading list, conceptual diagrams, and illustrative examples.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Institutional subscriptions

Abbreviations

BPAg1:

Bullous pemphigoid antigen 1

BPAg2:

Bullous pemphigoid antigen 2

DIF:

Direct immunofluorescence

EBA:

Epidermolysis bullosa acquisita

ELISA:

Enzyme-linked immunosorbent serum assay

kDa:

kilodaltons

OCT:

Optimal cutting temperature

References

  1. Michel B, Milner Y, David K. Preservation of tissue-fixed immunoglobulins in skin biopsies of patients with lupus erythematosus and bullous diseases – preliminary report. J Invest Dermatol. 1972;59(6):449–52.

    Article  PubMed  CAS  Google Scholar 

  2. Vodegel RM, de Jong MC, Meijer HJ, Weytingh MB, Pas HH, Jonkman MF. Enhanced diagnostic immunofluorescence using biopsies transported in saline. BMC Dermatol. 2004;4:10.

    Article  PubMed  Google Scholar 

  3. Lazarova Z, Yancey KB. Reactivity of autoantibodies from patients with defined subepidermal bullous diseases against 1 mol/L salt-split skin. Specificity, sensitivity, and practical considerations. J Am Acad Dermatol. 1996;35(31):398–403.

    Article  PubMed  CAS  Google Scholar 

  4. Vodegel RM, Jonkman MF, Pas HH, de Jong MC. U-serrated immunodeposition pattern differentiates type VII collagen targeting bullous diseases from other subepidermal bullous autoimmune diseases. Br J Dermatol. 2004;151(1):112–8.

    Article  PubMed  CAS  Google Scholar 

  5. Chan LS, Ahmed AR, Anhalt GJ, et al. The first international consensus on mucous membrane pemphigoid: definition, diagnostic criteria, pathogenic factors, medical treatment, and prognostic indicators. Arch Dermatol. 2002;138(3):370–9.

    Article  PubMed  Google Scholar 

  6. Chan LS. Human skin basement membrane in health and in autoimmune diseases. Front Biosci. 1997;2:d343–52.

    PubMed  CAS  Google Scholar 

  7. Zillikens D. Diagnosis of autoimmune bullous skin diseases. Clin Lab. 2008;54(11–12):491–503.

    PubMed  CAS  Google Scholar 

  8. Rose C, Weyers W, Denisjuk N, Hillen U, Zillikens D, Shimanovich I. Histopathology of anti-p200 pemphigoid. Am J Dermatopathol. 2007;29(2):119–24.

    Article  PubMed  Google Scholar 

  9. Mutasim DF, Bilic M, Hawayek LH, Pipitone MA, Sluzevich JC. Immunobullous diseases. J Am Acad Dermatol. 2005;52(6):1029–43.

    Article  PubMed  Google Scholar 

  10. Kasperkiewicz M, Zillikens D. The pathophysiology of bullous pemphigoid. Clin Rev Allergy Immunol. 2007;33(1–2):67–77.

    Article  PubMed  CAS  Google Scholar 

  11. Fleming TE, Korman NJ. Cicatricial pemphigoid. J Am Acad Dermatol. 2000;43(4):571–91.

    Article  PubMed  CAS  Google Scholar 

  12. Castro LA, Lundell RB, Krause PK, Gibson LE. Clinical experience in pemphigoid gestationis: report of 10 cases. J Am Acad Dermatol. 2006;55(5):823–8.

    Article  PubMed  Google Scholar 

  13. Jenkins RE, Hern S, Black MM. Clinical features and management of 87 patients with pemphigoid gestationis. Clin Exp Dermatol. 1999;24(4):255–9.

    Article  PubMed  CAS  Google Scholar 

  14. Chi CC, Wang SH, Charles-Holmes R, et al. Pemphigoid gestationis: early onset and blister formation are associated with adverse pregnancy outcomes. Br J Dermatol. 2009;160(6):1222–8.

    Article  PubMed  Google Scholar 

  15. Sitaru C, Dahnrich C, Probst C, et al. Enzyme-linked immunosorbent assay using multimers of the 16th non-collagenous domain of the BP180 antigen for sensitive and specific detection of pemphigoid autoantibodies. Exp Dermatol. 2007;16(9):770–7.

    Article  PubMed  CAS  Google Scholar 

  16. Kirtschig G, Marinkovich MP, Burgeson RE, Yancey KB. Anti-basement membrane autoantibodies in patients with anti-epiligrin cicatricial pemphigoid bind the alpha subunit of laminin 5. J Invest Dermatol. 1995;105(4):543–8.

    Article  PubMed  CAS  Google Scholar 

  17. Egan CA, Lazarova Z, Darling TN, Yee C, Yancey KB. Anti-epiligrin cicatricial pemphigoid: clinical findings, immunopathogenesis, and significant associations. Medicine (Baltimore). 2003;82(3):177–86.

    Google Scholar 

  18. Rose C, Schmidt E, Kerstan A, et al. Histopathology of anti-laminin 5 mucous membrane pemphigoid. J Am Acad Dermatol. 2009;61(3):433–40.

    Article  PubMed  CAS  Google Scholar 

  19. Pas HH. Immunoblot assay in differential diagnosis of autoimmune blistering skin diseases. Clin Dermatol. 2001;19(5):622–30.

    Article  PubMed  CAS  Google Scholar 

  20. Mihai S, Sitaru C. Immunopathology and molecular diagnosis of autoimmune bullous diseases. J Cell Mol Med. 2007;11(3):462–81.

    Article  PubMed  CAS  Google Scholar 

  21. Remington J, Chen M, Burnett J, Woodley DT. Autoimmunity to type VII collagen: epidermolysis bullosa acquisita. Curr Dir Autoimmun. 2008;10:195–205.

    Article  PubMed  CAS  Google Scholar 

  22. Chen M, Chan LS, Cai X, O’Toole EA, Sample JC, Woodley DT. Development of an ELISA for rapid detection of anti-type VII collagen autoantibodies in epidermolysis bullosa acquisita. J Invest Dermatol. 1997;108(1):68–72.

    Article  PubMed  CAS  Google Scholar 

  23. Kasperkiewicz M, Meier M, Zillikens D, Schmidt E. Linear IgA disease: successful application of immunoadsorption and review of the literature. Dermatology. 2010;220(3):259–63.

    Article  PubMed  CAS  Google Scholar 

  24. Waldman MA, Black DR, Callen JP. Vancomycin-induced linear IgA bullous disease presenting as toxic epidermal necrolysis. Clin Exp Dermatol. 2004;29(6):633–6.

    Article  PubMed  CAS  Google Scholar 

  25. Gruschwitz M, Keller J, Hornstein OP. Deposits of immunoglobulins at the dermo-epidermal junction in chronic light-exposed skin: what is the value of the lupus band test? Clin Exp Dermatol. 1988;13(5):303–8.

    Article  PubMed  CAS  Google Scholar 

  26. Kontos AP, Jirsari M, Jacobsen G, Fivenson DP. Immunoglobulin M predominance in cutaneous lupus erythematosus. J Cutan Pathol. 2005;32(5):352–5.

    Article  PubMed  Google Scholar 

  27. Crowson AN, Magro C. The cutaneous pathology of lupus erythematosus: a review. J Cutan Pathol. 2001;28(1):1–23.

    Article  PubMed  CAS  Google Scholar 

  28. Gammon WR, Woodley DT, Dole KC, Briggaman RA. Evidence that anti-basement membrane zone antibodies in bullous eruption of systemic lupus erythematosus recognize epidermolysis bullosa acquisita autoantigen. J Invest Dermatol. 1985;84(6):472–6.

    Article  PubMed  CAS  Google Scholar 

  29. David-Bajar KM, Bennion SD, DeSpain JD, Golitz LE, Lee LA. Clinical, histologic, and immunofluorescent distinctions between subacute cutaneous lupus erythematosus and discoid lupus erythematosus. J Invest Dermatol. 1992;99(3):251–7.

    Article  PubMed  CAS  Google Scholar 

  30. Sarkany RP. The management of porphyria cutanea tarda. Clin Exp Dermatol. 2001;26(3):225–32.

    Article  PubMed  CAS  Google Scholar 

  31. Egbert BM, LeBoit PE, McCalmont T, Hu CH, Austin C. Caterpillar bodies: distinctive, basement membrane-containing structures in blisters of porphyria. Am J Dermatopathol. 1993;15(3):199–202.

    Article  PubMed  CAS  Google Scholar 

  32. Green JJ, Manders SM. Pseudoporphyria. J Am Acad Dermatol. 2001;44(1):100–8.

    Article  PubMed  CAS  Google Scholar 

  33. Groves RWH. Pemphigus: a brief review. Clin Med. 2009;9(4):371–5.

    Article  PubMed  Google Scholar 

  34. Ishii K, Amagai M, Hall RP, et al. Characterization of autoantibodies in pemphigus using antigen-specific enzyme-linked immunosorbent assays with baculovirus-expressed recombinant desmogleins. J Immunol. 1997;159(4):2010–7.

    PubMed  CAS  Google Scholar 

  35. Amagai M, Komai A, Hashimoto T, et al. Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus. Br J Dermatol. 1999;140(2):351–7.

    Article  PubMed  CAS  Google Scholar 

  36. Dasher D, Rubenstein D, Diaz LA. Pemphigus foliaceus. Curr Dir Autoimmun. 2008;10:182–94.

    Article  PubMed  CAS  Google Scholar 

  37. Amerian ML, Ahmed AR. Pemphigus erythematosus. Presentation of four cases and review of literature. J Am Acad Dermatol. 1984;10(2 Pt 1):215–22.

    Article  PubMed  CAS  Google Scholar 

  38. Robinson ND, Hashimoto T, Amagai M, Chan LS. The new pemphigus variants. J Am Acad Dermatol. 1999;40(5 Pt 1):649–71.

    Article  PubMed  CAS  Google Scholar 

  39. Duker I, Schaller J, Rose C, Zillikens D, Hashimoto T, Kunze J. Subcorneal pustular dermatosis-type IgA pemphigus with autoantibodies to desmocollins 1, 2, and 3. Arch Dermatol. 2009;145(10):1159–62.

    Article  PubMed  Google Scholar 

  40. Ishii N, IshidaYamamoto A, Hashimoto T. Immunolocalization of target autoantigens in IgA pemphigus. Clin Exp Dermatol. 2004;29(1):62–6.

    Article  PubMed  CAS  Google Scholar 

  41. Anhalt GJ. Paraneoplastic pemphigus. J Investig Dermatol Symp Proc. 2004;9(1):29–33.

    Article  PubMed  Google Scholar 

  42. Kaplan I, Hodak E, Ackerman L, Mimouni D, Anhalt GJ, Calderon S. Neoplasms associated with paraneoplastic pemphigus: a review with emphasis on non-hematologic malignancy and oral mucosal manifestations. Oral Oncol. 2004;40(6):553–62.

    Article  PubMed  Google Scholar 

  43. Probst C, Schlumberger W, Stocker W, et al. Development of ELISA for the specific determination of autoantibodies against envoplakin and periplakin in paraneoplastic pemphigus. Clin Chim Acta. 2009;410(1–2):13–8.

    Article  PubMed  CAS  Google Scholar 

  44. Madan V, August PJ. Exophytic plaques, blisters, and mouth ulcers. Pemphigus vegetans (PV), Neumann type. Arch Dermatol. 2009;145(6):715–20.

    Article  PubMed  Google Scholar 

  45. Ma DL, Fang K. Hallopeau type of pemphigus vegetans confined to the right foot: case report. Chin Med J. 2009;122(5):588–90.

    PubMed  Google Scholar 

  46. Helander SD, Rogers 3rd RS. The sensitivity and specificity of direct immunofluorescence testing in disorders of mucous membranes. J Am Acad Dermatol. 1994;30(1):65–75.

    Article  PubMed  CAS  Google Scholar 

  47. Raghu AR, Nirmala NR, Sreekumaran N. Direct immunofluorescence in oral lichen planus and oral lichenoid reactions. Quintessence Int. 2002;33(3):234–9.

    PubMed  Google Scholar 

  48. Zillikens D, Caux F, Mascaro JM, et al. Autoantibodies in lichen planus pemphigoides react with a novel epitope within the C-terminal NC16A domain of BP180. J Invest Dermatol. 1999;113(1):117–21.

    Article  PubMed  CAS  Google Scholar 

  49. Cohen DM, Ben-Amitai D, Feinmesser M, Zvulunov A. Childhood lichen planus pemphigoides: a case report and review of the literature. Pediatr Dermatol. 2009;26(5):569–74.

    Article  PubMed  Google Scholar 

  50. Magro CM, Crowson AN. A clinical and histologic study of 37 cases of immunoglobulin A-associated vasculitis. Am J Dermatopathol. 1999;21(3):234–40.

    Article  PubMed  CAS  Google Scholar 

  51. Crowson AN, Mihm Jr MC, Magro CM. Cutaneous vasculitis: a review. J Cutan Pathol. 2003;30(3):161–73.

    Article  PubMed  Google Scholar 

  52. Magro CM, Crowson AN. The immunofluorescent profile of ­dermatomyositis: a comparative study with lupus erythematosus. J Cutan Pathol. 1997;24(9):543–52.

    Article  PubMed  CAS  Google Scholar 

  53. Zaenglein AL, Hafer L, Helm KF. Diagnosis of dermatitis herpetiformis by an avidin-biotin-peroxidase method. Arch Dermatol. 1995;131(5):571–3.

    Article  PubMed  CAS  Google Scholar 

  54. Chandler W, Zone J, Florell S. C4d immunohistochemical stain is a sensitive method to confirm immunoreactant deposition in formalin-fixed paraffin-embedded tissue in bullous pemphigoid. J Cutan Pathol. 2009;36(6):655–9.

    Article  PubMed  Google Scholar 

  55. Magro CM, Dyrsen ME. The use of C3d and C4d immunohistochemistry on formalin-fixed tissue as a diagnostic adjunct in the assessment of inflammatory skin disease. J Am Acad Dermatol. 2008;59(5):822–33.

    Article  PubMed  Google Scholar 

  56. Vodegel RM, de Jong MC, Pas HH, Jonkman MF. IgA-mediated epidermolysis bullosa acquisita: two cases and review of the literature. J Am Acad Dermatol. 2002 Dec;47(6):919–25. Review.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dermatopathology Section, Department of Laboratory Medicine, Geisinger Medical Center, 100 North Academy Avenue, Danville, PA, 17822, USA

    William B. Tyler

Authors
  1. William B. Tyler
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to William B. Tyler .

Editor information

Editors and Affiliations

  1. , Pathology and Laboratory Medicine, Geisinger Health System, 100 N. Academy Avenue, Danville, 17822, Pennsylvania, USA

    Fan Lin

  2. , Pathology and Laboratory Medicine, Geisinger Medical Center, 100 N. Academy Avenue, Danville, 17822, Pennsylvania, USA

    Jeffrey Prichard

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this chapter

Cite this chapter

Tyler, W.B. (2011). Application of Direct Immunofluorescence for Skin and Mucosal Biopsies: A Practical Review. In: Lin, F., Prichard, J. (eds) Handbook of Practical Immunohistochemistry. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-8062-5_31

Download citation

  • DOI: https://doi.org/10.1007/978-1-4419-8062-5_31

  • Published:

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4419-8061-8

  • Online ISBN: 978-1-4419-8062-5

  • eBook Packages: MedicineMedicine (R0)

Publish with us

Policies and ethics

Search

Navigation