| In the late 1970s, several proteins, most
notably bullous pemphigoid antigen-1, laminin-1, and type IV collagen, were shown to he
normal components of the dermoepidermal junction by the use of polyclonal antibodies
specific for these proteins and an indirect immunofluorescence staining technique.
In 1981, Hintner and colleagues demonstrated that these three different antibodies type
could be used to accurately define the level of skin cleavage present in EB skin,
thereby permitting proper diagnosis and classification. As will be discussed in greater
detail in chapter 3, this immunohistochemical technique, now
known as immunofluorescence antigenic mapping, continues to be used as a diagnostic
alternative to transmission electron microscopy. In the early
1980s, a variety of new antigens were detected in the dermoepidermal junction, using
monoclonal antibody technique. In December 1983, Goldsmith and Briggaman were the first to
use this technique to demonstrate altered staining of two as yet to be characterized
anchoring fibril-associated antigens, named AF1 and AF2, in RDEB skin. The following
month, Fine and colleagues reported on the semiquantitative alteration in staining of a
lamina densa-specific antigen, KF-1, in both RDEB and DDEB skin. Based on these collective
findings, both sets of monoclonal antibodies were then used as a means of diagnosing and
classifying DEB patients. In 1986, Heagerty, Eady, and colleagues demonstrated absence of
staining of RDEB skin with another antibasement membrane-specific monoclonal antibody, LH
7:2, which was later shown to recognize an epitope of type VII collagen. In the same year,
Heagerty and co-workers employed two other basement membrane-specific antibodies, GB3
(later shown to recognize laminin-5) and AA3, in the postnatal diagnosis of the Herlitz
subtype of generalized JEB. GB3, LH 7:2, and KF-1 monoclonal antibodies were subsequently
shown to he useful probes for the pre- natal diagnosis of Herlitz JEB and DEB subtypes,
respectively. In 1989, Fine and colleagues identified a new component of the anchoring
filament (now termed uncein) with the 19-DEJ-1 monoclonal antibody and then demonstrated
its lack of detection in any JEB skin, suggesting its superiority over GB3 as an
immunohistochemical probe for the detection of all JEB subtypes. In 1990, Schofield and
colleagues more precisely defined the sensitivity and specificity of GB3 as a diagnostic
marker of JEB. |
