マウス敗血症モデルを用い、微小循環における白血球-血小板-血管内皮細胞の相互作用を二光子励起顕微鏡下で生体内可視化した。盲腸壁内の後毛細管細静脈や肝類洞では、白血球の血管内皮細胞への接着、白血球への血小板接着、血小板凝集が観察できた。また、これらの反応で惹起されるNeutrophil Extracellular Traps(NETs)を可視化できた。NETsは形態学的に、anchored NETsとcell-free NETsに分類できた。組織微小血管内における白血球-血小板-血管内皮細胞の過剰な相互作用は循環障害を引き起こす可能性があり、NETsも関与している可能性が示唆された。
We developed a method of intravital imaging of interaction between leukocytes, platelets and vascular endothelium in tissue microcirculation of a lipopolysaccharide -induced sepsis model using a two photon laser scanning microscopy. We imaged the rolling or adhesion of leukocytes to vascular endothelium, leukocyte-platelet aggregates, and platelet aggregation in postcapillary venules of the cecum and hepatic sinusoids in vivo real-time. We also imaged neutrophil extracellular traps (NETs) which are activated by these interactions. NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils (fragmented or cotton-like structures) and anchored NETs that were anchored to neutrophils (linear, reticular, membranous, or spot-like structures). NETs seemed to be associated with the formation of platelet aggregates or leukocyte-platelet aggregates. These observations may suggest the adverse effect of intravascular NETs on the host during a sepsis.
二光子励起顕微鏡を用いた敗血症マウス微小循環障害の生体内リアルタイムイメージング
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