Cytological characterization of apatitic calcium phosphate structures in bronchial epithelial tissue cultured from a child with cystic fibrosis (deltaF508).

Abstract

Patients with cystic fibrosis homozygous for the AF508 mutation have marked disturbances in ion exchanges in a variety of tissues. Alterations in intra- or extracellular levels of Ca2+ and calcifications have been observed in numerous tissues from such patients, although the nature and origin of such calcifications have yet to be elucidated. In this study, we investigated the formation of calcifications in the respiratory tract of a AF508 homozygous child and attempted to establish their origin. Samples of bronchial epithelium from this patient were subjected to cytophysiological analysis ex vivo and in vitro. The defect of targeting of the cystic fibrosis transmembrane conductance regulator (CFTR) to the apical plasma membrane of epithelial cells was verified. Cytochemical and ultrastructural analysis revealed the presence of crystalline aggregates in fine needles in the respiratory tract. Cytochemical analysis, X-ray spectrometry, and electron diffraction showed that these aggregates corresponded to crystals of calcium phosphate in an apatite-like structure. Ultrastructural study of primary cultures of bronchial epithelium showed the presence of calcium phosphate crystals in granules from Golgi apparatus and in mitochondria. These observations indicated that modifications of ionic exchanges due to a defect in targeting of CFTR AF508 to the apical plasma membrane led to the formation of crystals of calcium phosphate in the cytoplasm of pulmonary cells. These crystals could enhance inflammation of the lung in patients with cystic fibrosis.