S. Kumari et al., Identification of ocular cicatricial Pemphigoid antibody binding site(s) in human beta 4 integrin, INV OPHTH V, 42(2), 2001, pp. 379-385
PURPOSE. To identify specific site(s) on human beta4 molecule to which sera
from ocular cicatricial pemphigoid (OCP) patients bind and to determine it
s role in the process of blister formation.
METHODS. Clone the fragments representing the extracellular and intracellul
ar domain of beta4 molecule from normal human conjunctival mRNA into an exp
ression vector; map the region to which sera from OCP patients bind by West
ern blot analysis. Determine the role of the immunodominant region in patho
genesis by demonstrating the ability of the rabbit antibody to the immunodo
minant region to produce separation of basement membrane zone (BMZ) from th
e basal epithelial layer when incubated with normal human conjunctiva in an
in vitro organ culture model.
RESULTS. Majority of the OCP sera tested bound to the C-terminal end of the
intracellular domain IC3.0 of the human beta4 integrin. Further subcloning
of IC3.0 demonstrated that a smaller fragment extending from 1489 aa to 15
72 aa (IC3.4) was responsible for this binding. This region may have multip
le antibody binding sites. Antibody to human IC3.0 and IC3.4 produced in ra
bbit, resulted in BMZ separation, histologically identical with that observ
ed when normal human conjunctiva was cultured with OCP sera in an human con
junctival organ culture model.
CONCLUSIONS. These observations identify IC3.4 as the antibody binding site
for sera of OCP patients and suggest a possible role for it in blister for
mation. Indirectly it highlights certain important aspects of the structura
l and functional dynamics of the biology of the hemidesmosomes and basement
membranes.