Applicability of different antibodies for immunohistochemical localizationof CFTR in sweat glands from healthy controls and from patients with cystic fibrosis
A. Claass et al., Applicability of different antibodies for immunohistochemical localizationof CFTR in sweat glands from healthy controls and from patients with cystic fibrosis, J HIST CYTO, 48(6), 2000, pp. 831-837
The hereditary disease cystic fibrosis (CF) is caused by mutations in the c
ystic fibrosis transmembrane conductance regulator (CFTR) gene. Understandi
ng of the consequences of CFTR gene mutations is derived chiefly from in vi
tro studies on heterologous cell cultures and on cells hyperexpressing CFTR
. Data from ex vivo studies on human tissue are scarce and contradictory, a
fact which is in part explained by secondary tissue destruction in most af
fected organs. The purpose of this study was to establish conditions under
which wild-type and mutated CFTR can be studied in affected human tissue. S
weat glands carry the basic defect underlying CF and are not affected by ti
ssue destruction and inflammation. Therefore, we used this tissue to test a
panel of eight different CFTR antibodies under various fixation techniques
. The antibodies were tested on skin biopsy sections from healthy controls,
from CF patients homozygous for the most common mutation, Delta F508, and
from patients carrying two nonsense mutations. Of the eight CFTR antibodies
, only three-M3A7, MATG 1104, and cc24-met the criteria necessary for immun
olocalization of CFTR in sweat glands. The labeling pattern in the CF sweat
glands was consistent with the postulated processing defect: of Delta F508
CFTR. The antibodies exhibited different sensitivities for detecting Delta
F508 CFTR.