Schneider T, van Velzen D, Moqbel R, Issekutz A C
Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia, Canada.
Am J Respir Cell Mol Biol. 1997 Dec;17(6):702-12. doi: 10.1165/ajrcmb.17.6.2849.
We quantitated neutrophil and eosinophil migration into lung parenchyma using specific peroxidase enzyme assays, and into the bronchoalveolar compartment by bronchoalveolar lavage (BALF), in sensitized brown Norway (BN), Fischer, and Lewis rats and also assessed the lungs by histopathology. Fourteen days after sensitization with ovalbumin (OA in alum [given subcutaneously] and OA with Bordetella pertussis [given intraperitoneally]), rats were challenged with an OA aerosol for 1 h. In BN rats, there was marked perivascular and peribronchial edema, focal hemorrhages, and increase in lung wet weight and BALF protein content, accompanied by neutrophilic infiltration at 3-14 h postchallenge. Few eosinophils were seen at 14 h in lung tissue or in BALF. Neutrophils peaked at 24 h in parenchyma ([94 +/- 7] x 10[6]) and in BALF ([2.7 +/- 0.4] x 10[6]) and declined rapidly thereafter. Marked eosinophil infiltration into parenchyma was apparent by 24 h. Eosinophil accumulation peaked at 48 h in parenchyma ([127 +/- 18] x 10[6]) and at 72 h in BALF ([10 +/- 2.4] x 10[6]), comprising up to 85% of lavage cells at this time. Lung eosinophilia persisted for at least 6 d with only a slow decline or clearance, not approximating baseline until day 13 after challenge. Histopathology showed peribronchial and interstitial eosinophilic pneumonia, most severe on day 3. In contrast to the BN rats, essentially no pulmonary inflammation was observed in Lewis and Fischer rats. This model in the BN rat, and the specific peroxidase assays for quantitating tissue eosinophils and neutrophils, should be useful for investigating the regulation of allergen-induced eosinophil and neutrophil migration into and clearance from the lung.
我们使用特定的过氧化物酶测定法对中性粒细胞和嗜酸性粒细胞向肺实质的迁移进行定量,并通过支气管肺泡灌洗(BALF)对其向支气管肺泡腔的迁移进行定量,实验对象为致敏的棕色挪威(BN)大鼠、Fischer大鼠和Lewis大鼠,同时还通过组织病理学对肺进行评估。在用卵清蛋白(皮下注射明矾中的OA和腹腔注射百日咳博德特氏菌中的OA)致敏14天后,用OA气雾剂对大鼠进行1小时的激发。在BN大鼠中,激发后3至14小时出现明显的血管周围和支气管周围水肿、局灶性出血、肺湿重增加和BALF蛋白含量增加,伴有中性粒细胞浸润。在14小时时,在肺组织或BALF中很少见到嗜酸性粒细胞。中性粒细胞在24小时时在实质中([94±7]×10⁶)和BALF中([2.7±0.4]×10⁶)达到峰值,此后迅速下降。到24小时时,明显可见嗜酸性粒细胞大量浸润到实质中。嗜酸性粒细胞在实质中的积聚在48小时时达到峰值([127±18]×10⁶),在BALF中的积聚在72小时时达到峰值([10±2.4]×10⁶),此时占灌洗细胞的比例高达85%。肺嗜酸性粒细胞增多持续至少6天,仅缓慢下降或清除,直到激发后第13天才接近基线水平。组织病理学显示支气管周围和间质性嗜酸性粒细胞肺炎,在第3天最为严重。与BN大鼠相反,在Lewis大鼠和Fischer大鼠中基本未观察到肺部炎症。BN大鼠的这个模型以及用于定量组织嗜酸性粒细胞和中性粒细胞的特定过氧化物酶测定法,对于研究变应原诱导的嗜酸性粒细胞和中性粒细胞向肺内迁移以及从肺内清除的调节应该是有用的。