Cooled neutrophils become deposited in the microcirculation after infusion: A potential mechanism for microvascular disruption following tissue hypothermia

Abstract

1. We examined whether changes in rigidity and adhesiveness of neutrophils exposed to cooling and rewarming observed in vitro might impair microvascular perfusion in vivo. Neutrophils from donor rats were fluorescently (calcein-AM) or radioactively (Indium-111) labelled, incubated at 10 or 37 °C in vitro, and infused into recipients. Changes in transit rate and adhesive behaviour within post-capillary venules was quantified in m. extensor digitorum longus (EDL) using intravital microscopy, and tissue distribution determined. 2. There was an increased propensity of cooled cells to undergo adhesion following transfer into the recipient rat. However, cooling had no effect on median transit (354 μm s −1) or rolling (14 μm s −1) velocities during the first 5 min after infusion suggesting that cooling promotes adhesion, but does not delay passage through capillaries. Cooled neutrophils subsequently transformed to stationary adhesion. Their immobilisation was higher than for cells held at 37 °C ( P<0.05), and once immobilised they remained firmly adherent to the vessel wall. Cooled, radiolabelled neutrophils showed tissue-specific accumulation after 3 min, but were cleared to the same extent as warmed cells by 20 min. 3. Our data suggest that cooling and rewarming of neutrophils impairs their ability to transit microvessels, reflecting changes in adhesive and mechanical properties observed in vitro, and may contribute to cold-associated circulatory pathology.