ovalbumin and Cystic-Fibrosis

Topics: Amyloidosis; Animals; Antigens; Basement Membrane; Blood Group Antigens; Brain Chemistry; Carbohydrates; Cell Membrane; Collagen; Connective Tissue; Cystic Fibrosis; Diabetic Angiopathies; Enzymes; gamma-Globulins; Glycoproteins; Hormones; Humans; Mucins; Mucous Membrane; Neoplasms; Ovalbumin

Inflammatory airway diseases, such as asthma, cystic fibrosis (CF), and chronic obstructive pulmonary disease (COPD), are characterized by mucus hypersecretion and airway plugging. In both CF and asthma, enhanced expression of the Ca2+-activated Cl- channel TMEM16A is detected in mucus-producing club/goblet cells and airway smooth muscle. TMEM16A contributes to mucus hypersecretion and bronchoconstriction, which are both inhibited by blockers of TMEM16A, such as niflumic acid. Here we demonstrate that the FDA-approved drug niclosamide, a potent inhibitor of TMEM16A identified by high-throughput screening, is an inhibitor of both TMEM16A and TMEM16F. In asthmatic mice, niclosamide reduced mucus production and secretion, as well as bronchoconstriction, and showed additional antiinflammatory effects. Using transgenic asthmatic mice, we found evidence that TMEM16A and TMEM16F are required for normal mucus production/secretion, which may be due to their effects on intracellular Ca2+ signaling. TMEM16A and TMEM16F support exocytic release of mucus and inflammatory mediators, both of which are blocked by niclosamide. Thus, inhibition of mucus and cytokine release, bronchorelaxation, and reported antibacterial effects make niclosamide a potentially suitable drug for the treatment of inflammatory airway diseases, such as CF, asthma, and COPD.

Topics: Animals; Anoctamins; Anti-Inflammatory Agents; Asthma; Bronchi; Cell Line, Tumor; Cystic Fibrosis; Disease Models, Animal; Drug Repositioning; Goblet Cells; HEK293 Cells; Humans; Mice; Mice, Knockout; Mice, Transgenic; Mucus; Niclosamide; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Signal Transduction

Highly viscous mucus poses a big challenge for the delivery of particulates carrying therapeutics to patients with cystic fibrosis. In this study, surface modifying DNase I loaded particles using different excipients to achieve better lung deposition, higher enzyme stability or better biological activity had been exploited. For the purpose, controlled release microparticles (MP) were prepared by co-spray drying DNase I with the polymer poly-lactic-co-glycolic acid (PLGA) and the biocompatible lipid surfactant 1,2-dipalmitoyl-Sn-phosphatidyl choline (DPPC) using various hydrophilic excipients. The effect of the included modifiers on the particle morphology, size, zeta potential as well as enzyme encapsulation efficiency, biological activity and release had been evaluated. Powder aerosolisation performance and particle phagocytosis by murine macrophages were also investigated. The results showed that more than 80% of enzyme activity was recovered after MP preparation and that selected surface modifiers greatly increased the enzyme encapsulation efficiency. The particle morphology was greatly modified altering in turn the powders inhalation indices where dextran, ovalbumin and chitosan hydrochloride increased considerably the respirable fraction compared to the normal hydrophilic carriers lactose and PVP. Despite of the improved aerosolisation caused by chitosan hydrochloride, yet retardation of chitosan coated particles in artificial mucus samples discouraged its application. On the other hand, dextran and polyanions enhanced DNase I effect in reducing cystic fibrosis mucus viscosity. DPPC proved good ability to reduce particles phagocytic uptake even in the presence of the selected adjuvants. The prepared MP systems were biocompatible with lung epithelial cells. To conclude, controlled release DNase I loaded PLGA-MP with high inhalation indices and enhanced mucolytic activity on CF sputum could be obtained by surface modifying the particles with PGA or dextran.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Administration, Inhalation; Aerosols; Animals; Chitosan; Cystic Fibrosis; Delayed-Action Preparations; Deoxyribonuclease I; Dextrans; Drug Carriers; Drug Delivery Systems; Epithelial Cells; Excipients; Lactic Acid; Macrophages; Mice; Mucus; Ovalbumin; Polyglutamic Acid; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Powders; Viscosity

Topics: Complement C3; Cystic Fibrosis; Electrophoresis, Agar Gel; Fluorescent Antibody Technique; Humans; Hypersensitivity; Immunoglobulin A, Secretory; Immunoglobulin E; Ovalbumin; Precipitins; Saliva; Serum Albumin, Bovine; Sputum

Homogenates of cultivated skin fibroblasts derived from patients with cystic fibrosis had a higher level of UDP-galactose:ovalbumin galactosyl transferase activity compared to fibroblasts derived from control subjects. The activity in control subjects was 1.82 +/- 0.43 nmol galactose transferred/hr/mg protein, whereas the activity in fibroblasts of patients was 2.95 +/- 0.77. The difference was significant at P less than 0.01. Activity in the fibroblasts of obligate heterozygotes was 2.15 +/- 0.60. The difference between the activities in fibroblasts of heterozygotes and patients was significant at P less than 0.05. The activity in control fibroblasts could be enhanced by basic polypeptides like polylysine, polyarginine, histone, and protamine but not by neutral or acidic polypeptides. Fibroblasts from patients released significantly higher amounts of a soluble form of the enzyme activity into the culture medium than control fibroblasts.

Topics: Cells, Cultured; Culture Media; Cystic Fibrosis; Enzyme Activation; Fibroblasts; Galactosyltransferases; Heterozygote; Homozygote; Humans; Ovalbumin; Peptides; Skin; Uridine Diphosphate Galactose