Quantitative microscopy in murine models of lung inflammation.

OBJECTIVE: To develop a quantitative means to measure lung inflammation using the murine models of chronic asthma and cystic fibrosis (CF). STUDY DESIGN: Translational-based medicine often utilizes animal models to study new and innovative therapeutics. In asthma and CF, the animal models focus on airway inflammation and remodeling. The asthma model is based on hypersensitivity-induced airway disease, whereas the CF model focuses on the inflammatory response to infection with Pseudomonas aeruginosa. Qualitative measures of inflammation and lung pathophysiology introduce significant variability and difficulty in interpreting interventional outcomes. The highly sensitive and reproducible quantitative computational program interfaced with Image Pro Microscopy to monitor changes in lung inflammation and lung pathophysiology. The software interfaces with image microscopy and automates the lung section review process. RESULTS: Results from this program recapitulated data obtained by manual point counting of inflammation, bronchoalveolar lavage differential, and histology. The data show a low coefficient of variation and high reproducibility between slides and sections. CONCLUSION: Utilization of this new microscopy program will enhance the quantitative means of establishing changes in lung structure and inflammation as a measure of therapeutic intervention with the ability of refining interpretation of in vivo models potentially short-circuiting translation into the clinical setting.

A role for CFTR in the elevation of glutathione levels in the lung by oral glutathione administration.

The cystic fibrosis transmembrane conductance regulator (CFTR) protein is the only known apical glutathione (GSH) transporter in the lung. The purpose of these studies was to determine whether oral GSH or glutathione disulfide (GSSG) treatment could increase lung epithelial lining fluid (ELF) GSH levels and whether CFTR plays a role in this process. The pharmacokinetic profile of an oral bolus dose of GSH (300 mg/kg) was determined in mice. Plasma, ELF, bronchoalveolar lavage (BAL) cells, and lung tissue were analyzed for GSH content. There was a rapid elevation in the GSH levels that peaked at 30 min in the plasma and 60 min in the lung, ELF, and BAL cells after oral GSH dosing. Oral GSH treatment produced a selective increase in the reduced and active form of GSH in all lung compartments examined. Oral GSSG treatment (300 mg/kg) resulted in a smaller increase of GSH levels. To evaluate the role of CFTR in this process, Cftr knockout (KO) mice and gut-corrected Cftr KO-transgenic (Tg) mice were given an oral bolus dose of GSH (300 mg/kg) and compared with wild-type mice for changes in GSH levels in plasma, lung, ELF, and BAL cells. There was a twofold increase in plasma, a twofold increase in lung, a fivefold increase in ELF, and a threefold increase in BAL cell GSH levels at 60 min in wild-type mice; however, GSH levels only increased by 40% in the plasma, 60% in the lung, 50% in the ELF, and twofold in the BAL cells within the gut-corrected Cftr KO-Tg mice. No change in GSH levels was observed in the uncorrected Cftr KO mice. These studies suggest that CFTR plays an important role in GSH uptake from the diet and transport processes in the lung.