The early allergic response in small airways of human precision-cut lung slices

Abstract

To study the role of small airways in the early allergic response (EAR), the method of human precision-cut lung slices (PCLS) was developed and used to examine the bronchoconstriction elicited by passive sensitisation and allergen provocation.Viable human PCLS of 250-mm thickness containing airways v1.5 mm in outer diameter were prepared from lung lobes obtained from lung resection and taken into culture. According to the low release of lactate dehydrogenase and the constant ciliary beat frequency, human PCLS were viable for at least 3 days.Following overnight passive sensitisation with serum from allergic individuals, administration of grass-pollen extract or activating immunoglobulin E antibody resulted in immediate airway contraction that was quantified by videomicroscopy. The extent of the EAR increased with decreasing airway size (outer airway diameter), with the strongest response occurring in the terminal bronchioles.Histamine receptor antagonism was ineffective, and leukotriene or thromboxane receptor antagonism attenuated the early allergic response only in some cases. However, simultaneous blockade of leukotriene and thromboxane receptors almost completely prevented the early allergic response in the precision-cut lung slices from all individuals, suggesting such a dual treatment as a potential future asthma therapy.

Comparison Of Airway Responses In Sheep Of Different Age In Precision-cut Lung Slices (PCLS)

“…The microarchitecture of lung tissue, i.e. airways and vessels embedded in the parenchyma, remains intact and permit physiological measurements [23][24][25][26]. Even nerves remain functional and can be specifically activated [29].…”

“…Given the importance of small airways for asthma and COPD [24,25,32,33], the possibility to study these airways in PCLS is of clinical relevance. Small airways are defined by their inner diameter being less than 2 mm in human (airway generations .11) and less than 780 mm in rat airways [25,34].…”

“…PCLS were prepared from adult or newborn sheep lung as described in previous studies [14,17,[23][24][25] with some modifications. Briefly, the lungs were filled via the lobular bronchus with 1.5% low-melting-point agarose solution and put onto ice until the agarose had solidified.…”

“…PCLS can easily be prepared from different species and are already established for many species including rat, mouse, guinea pig, non-human primates and humans [14,17,[23][24][25][26]. PCLS represent a highly useful model to study bronchial and pulmonary vascular responses by videomicroscopy [23,27].…”

In Vitro Models of Chronic Obstructive Pulmonary Disease (COPD)

“…Unlike many continuous cell lines, lung slices retain the composite metabolic activity of the lung parenchyma and can be used in studies requiring bioactivation (Freeman & O'Neil 1984). Relatively large numbers of slices, up to 30 from one resection, can be generated from each donor (Wohlsen et al, 2003). However, large variation exists between sequential slices and also from donor to donor.…”