Immunochemical and Immunohistochemical Evaluation of Lung Permeability in Ventilated Newborn Rabbits

Abstract

These experiments were designed to quantify the vascular-to-alveolar leakage of albumin in the neonatal lung and to analyze the distribution of leaking airspaces in the lung parenchyma. Immediately after delivery, newborn rabbits with gestational age 27-29 days received an intravenous injection of human albumin as a marker and were ventilated for 15 min with standardized tidal volume (10 ml/kg). After the period of ventilation the lungs were either lavaged via the airways or fixed for histological studies. The median amount of albumin in lung lavage fluid, determined by immunodiffusion, was 4.8% of the injected dose after 27 days, 1.3% after 28 days, and 0.4% after 29 days of gestation; it was inversely correlated with the compliance of the respiratory system (r = –0.78; p < .001). Immunohistochemical examination of lung section revealed that the leak was not diffuse; even in animals with gestational age 27 days it involved only a median of 48% of total alveoli. The median amount of alveoli containing the label fell to 6% after 28 days and to 0% after 29 days gestation, correlating inversely with the compliance of the respiratory system (r = – 0.53; p < 0.01). We suggest that our experimental model is useful for histological demonstration of serum proteins leaking into the airspaces under experimental conditions and for evaluating the effect of therapeutic regiments on neonatal lung permeability.

Key Words:

• Immature lung
• lung development
• protein leakage
• artificial ventilation
• lung compliance
• immunodiffusion
• immunohistochemistry