Expression profiles of hydrophobic surfactant proteins in children with diffuse chronic lung disease

Abstract

Background: Abnormalities of the intracellular metabolism of the hydrophobic surfactant proteins SP-B and SP-C and their precursors may be causally linked to chronic childhood diffuse lung diseases. The profile of these proteins in the alveolar space is unknown in such subjects.

Genotype alone does not predict the clinical course ofSFTPCdeficiency in paediatric patients

“…[3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] (table 3). Empiric therapy included neonatal surfactant, corticosteroids (continuous and pulse therapy), colchicine, hydroxychloroquine, antibiotic therapy including azithromycin and whole lung lavage for initial alveolar proteinosis presentation.…”

“…SP-B and SP-C were determined as described [17,18]: proteins were separated on NuPage 10%; Bis-Tris gels (Novex X-cell Ii Mini Cell System; Novex, San Diego, CA, USA), transferred onto nitrocellulose membranes by Western blot in Nupage Blot modules (Novex) and incubated with antibodies as described [17]. For quantification, two standards were run on each gel; SP-B at 10 and 20 ng and SP-C at 10 and 25 ng.…”

Surfactant proteins in pediatric interstitial lung disease

“…SP-B and SP-C were determined as described previously (16,31). Briefly, SP-B and SP-C were separated on 10% Bis-Tris gels, identified by western blotting (using SP-C antibodies (generated in the rabbit against recombinant human SP-C, charge 22/96) and SP-B antibodies (generated in the rabbit against recombinant human SP-B, charge C 329); both gifts from Byk-Gulden, Konstanz, Germany) and quantified with the help of two standard runs on each gel (10 and 20 ng of SP-B, 10 and 25 ng of SP-C, using the Diana III chemiluminescence detection system and Advanced Image Data Analyzer software, version 4.04.032 (Raytest, Straubenhardt, Germany)) (Supplementary Figure S5 online).…”

“…From the previous studies, we know that there are qualitative differences in the expression profiles of SP-B and SP-C in the bronchoalveolar lavage fluids (BALF) of such children (16). However, not much is known regarding the quantitative levels of these proteins.…”

Surfactant Dysfunction in SP-A−/− and iNOS−/− Mice with Mycoplasma Infection

“…SP-B in cell-free BAL was detected as described by Griese and colleagues (25). For SP-B detection, the protein content each sample was determined with the BioRad Protein Assay Kit (BioRad, Richmond, CA) based on the method of Bradford (26).…”