Advertisement

Indirect Immunofluorescence Microscopy

  • Gilles F. H. DiercksEmail author
  • Hendri H. Pas
Chapter

Abstract

The purpose of indirect immunofluorescence microscopy is to detect circulating antibodies in patient’s serum. For this purpose, an adequate substrate is necessary to visualize these antibodies. Monkey esophagus is the most widely used substrate for detecting circulating autoantibodies in patients with autoimmune bullous diseases. In all variants of pemphigus, antibodies show an epithelial cell surface pattern, resulting from present autoantibodies against the desmosomal molecules desmoglein 1 and/or 3. This pattern is also called chicken wire or honeycomb pattern. In pemphigoid, a linear deposition along the epithelial basement membrane can be observed, caused by autoantibodies against hemidesmosomes or their connecting proteins underneath.

Human salt-split skin is a valuable substrate in the diagnosis of subepidermal autoimmune bullous diseases. Important antigens in the roof of salt-split skin are type XVII collagen (BP180) and BP230, whereas laminin 332, p200, and type IV collagen are situated in the floor of the blister. This implies that bullous pemphigoid, mucous membrane pemphigoid, pemphigoid gestationis, and lichen planus pemphigoides show staining of IgG on the epidermal side of the blister. On the other hand, anti-laminin 332 pemphigoid, anti-p200 pemphigoid, epidermolysis bullosa acquisita, and bullous SLE show staining on the dermal side.

Other less used, but valuable substrates in some instances, are rat bladder and knock-out skin.

Keywords

Immunofluorescence Pemphigus Pemphigoid Dermatitis herpetiformis 

References

  1. 1.
    Beutner EH, Jordon RE. Demonstration of skin antibodies in sera of pemphigus vulgaris patients by indirect immunofluorescent staining. Proc Soc Exp Biol Med. 1964;117:505–10.CrossRefPubMedGoogle Scholar
  2. 2.
    Feibelman C, Stolzner G, Provost TT. Pemphigus vulgaris. Superior sensitivity of monkey esophagus in the determination of pemphigus antibody. Arch Dermatol. 1981;117:561–2.CrossRefPubMedGoogle Scholar
  3. 3.
    Goldblatt F, Gordon TP. Antibodies to blood group antigens mimic pemphigus staining patterns: a useful reminder. Autoimmunity. 2002;35:93–6.CrossRefPubMedGoogle Scholar
  4. 4.
    Jordon RE, Beutner EH, Witebsky E, Blumental G, Hale WL, Lever WF. Basement zone antibodies in bullous pemphigoid. JAMA. 1967;200:751–6.CrossRefPubMedGoogle Scholar
  5. 5.
    Bean SF. Cicatricial pemphigoid. Immunofluorescent studies. Arch Dermatol. 1974;110(4):552–5.Google Scholar
  6. 6.
    Terra JB, Jonkman MF, Diercks GF, Pas HH. Low sensitivity of type VII collagen enzyme-linked immunosorbent assay in epidermolysis bullosa acquisita: serration pattern analysis on skin biopsy is required for diagnosis. Br J Dermatol. 2013;169:164–7.CrossRefPubMedGoogle Scholar
  7. 7.
    Jaremko WM, Beutner EH, Kumar V, Kipping H, Condry P, Zeid MY, Kauffmann CL, Tatakis DN, Chorzelski TP. Chronic ulcerative stomatitis associated with a specific immunologic marker. J Am Acad Dermatol. 1990;22:215–20.CrossRefPubMedGoogle Scholar
  8. 8.
    Chorzelski TP, Beutner EH, Sulej J, Tchorzewska H, Jablonska S, Kumar V, Kapuscinska A. IgA anti-endomysium antibody. A new immunological marker of dermatitis herpetiformis and coeliac disease. Br J Dermatol. 1984;111:395–402.CrossRefPubMedGoogle Scholar
  9. 9.
    Woodley D, Sauder D, Talley MJ, Silver M, Grotendorst G, Qwarnstrom E. Localization of basement membrane components after dermal-epidermal junction separation. J Invest Dermatol. 1983;81:149–53.CrossRefPubMedGoogle Scholar
  10. 10.
    Poot AM, Diercks GF, Kramer D, Schepens I, Klunder G, Hashimoto T, Borradori L, Jonkman MF, Pas HH. Laboratory diagnosis of paraneoplastic pemphigus. Br J Dermatol. 2013;169:1016–24.CrossRefPubMedGoogle Scholar
  11. 11.
    Vodegel RM, de Jong MC, Pas HH, Yancey KB, Jonkman MF. Anti-epiligrin cicatricial pemphigoid and epidermolysis bullosa acquisita: differentiation by use of indirect immunofluorescence microscopy. J Am Acad Dermatol. 2003;48:542–7.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2016

Authors and Affiliations

  1. 1.Departments of Pathology and Medical BiologyCenter for Blistering Diseases, University Medical Center Groningen, University of GroningenGroningenthe Netherlands
  2. 2.Department of DermatologyCenter for Blistering Diseases, University Medical Center Groningen, University of GroningenGroningenthe Netherlands

Personalised recommendations