Nita Izabela, Hollander Camilla, Westin Ulla, Janciauskiene Sabina-Marija
Department of Medicine, Lund University, University Hospital Malmö, 20502 Malmö, Sweden.
Respir Res. 2005 Jan 31;6(1):12. doi: 10.1186/1465-9921-6-12.
Alpha1-antitrypsin (AAT) serves primarily as an inhibitor of the elastin degrading proteases, neutrophil elastase and proteinase 3. There is ample clinical evidence that inherited severe AAT deficiency predisposes to chronic obstructive pulmonary disease. Augmentation therapy for AAT deficiency has been available for many years, but to date no sufficient data exist to demonstrate its efficacy. There is increasing evidence that AAT is able to exert effects other than protease inhibition. We investigated whether Prolastin, a preparation of purified pooled human AAT used for augmentation therapy, exhibits anti-bacterial effects.
Human monocytes and neutrophils were isolated from buffy coats or whole peripheral blood by the Ficoll-Hypaque procedure. Cells were stimulated with lipopolysaccharide (LPS) or zymosan, either alone or in combination with Prolastin, native AAT or polymerised AAT for 18 h, and analysed to determine the release of TNFalpha, IL-1beta and IL-8. At 2-week intervals, seven subjects were submitted to a nasal challenge with sterile saline, LPS (25 microg) and LPS-Prolastin combination. The concentration of IL-8 was analysed in nasal lavages performed before, and 2, 6 and 24 h after the challenge.
In vitro, Prolastin showed a concentration-dependent (0.5 to 16 mg/ml) inhibition of endotoxin-stimulated TNFalpha and IL-1beta release from monocytes and IL-8 release from neutrophils. At 8 and 16 mg/ml the inhibitory effects of Prolastin appeared to be maximal for neutrophil IL-8 release (5.3-fold, p < 0.001 compared to zymosan treated cells) and monocyte TNFalpha and IL-1beta release (10.7- and 7.3-fold, p < 0.001, respectively, compared to LPS treated cells). Furthermore, Prolastin (2.5 mg per nostril) significantly inhibited nasal IL-8 release in response to pure LPS challenge.
Our data demonstrate for the first time that Prolastin inhibits bacterial endotoxin-induced pro-inflammatory responses in vitro and in vivo, and provide scientific bases to explore new Prolastin-based therapies for individuals with inherited AAT deficiency, but also for other clinical conditions.
α1-抗胰蛋白酶（AAT）主要作为弹性蛋白降解蛋白酶、中性粒细胞弹性蛋白酶和蛋白酶3的抑制剂。有充分的临床证据表明，遗传性严重AAT缺乏易患慢性阻塞性肺疾病。AAT缺乏的增强疗法已经应用多年，但迄今为止尚无足够数据证明其疗效。越来越多的证据表明，AAT除了具有蛋白酶抑制作用外，还能发挥其他作用。我们研究了用于增强疗法的纯化人AAT制剂普洛莱斯坦（Prolastin）是否具有抗菌作用。
通过Ficoll-Hypaque法从血沉棕黄层或全外周血中分离人单核细胞和中性粒细胞。细胞用脂多糖（LPS）或酵母聚糖单独或与普洛莱斯坦、天然AAT或聚合AAT联合刺激18小时，并进行分析以确定肿瘤坏死因子α（TNFα）、白细胞介素-1β（IL-1β）和白细胞介素-8（IL-8）的释放。每隔2周，7名受试者接受无菌生理盐水、LPS（25微克）和LPS-普洛莱斯坦组合的鼻腔激发试验。在激发试验前以及激发后2、6和24小时进行的鼻腔灌洗中分析IL-8的浓度。
在体外，普洛莱斯坦对单核细胞内毒素刺激的TNFα和IL-1β释放以及中性粒细胞IL-8释放表现出浓度依赖性（0.5至16毫克/毫升）抑制作用。在8和16毫克/毫升时，普洛莱斯坦对中性粒细胞IL-8释放（与酵母聚糖处理的细胞相比，抑制5.3倍，p<0.001）以及单核细胞TNFα和IL-1β释放（与LPS处理的细胞相比，分别抑制10.7倍和7.3倍，p<0.001）的抑制作用似乎最大。此外，普洛莱斯坦（每侧鼻孔2.5毫克）显著抑制了对纯LPS激发的鼻腔IL-8释放。
我们的数据首次证明普洛莱斯坦在体外和体内抑制细菌内毒素诱导的促炎反应，并为探索基于普洛莱斯坦的新疗法提供了科学依据，不仅适用于遗传性AAT缺乏的个体，也适用于其他临床情况。
