Smith P D, Janoff E N, Mosteller-Barnum M, Merger M, Orenstein J M, Kearney J F, Graham M F
Department of Medicine, University of Alabama School of Medicine, Birmingham 35294, USA.
J Immunol Methods. 1997 Mar 10;202(1):1-11. doi: 10.1016/s0022-1759(96)00204-9.
Mucosal macrophages play a fundamental role in the regulation of immunological events and inflammation in the small intestine. Because no information is available on normal small intestinal macrophages, we developed a technique for the isolation and purification of jejunal lamina propria macrophages in order to study their phenotype and activity. From sections of normal human jejunum, lamina propria mononuclear cells were isolated by neutral protease digestion and then subjected to counterflow centrifugal elutriation to purify the macrophages. The cells isolated by this procedure contained < 1% CD3+ lymphocytes and displayed the size distribution, morphological features, ultrastructure and phagocytic activity of mononuclear phagocytes. In contrast to blood monocytes, however, mucosal macrophages from the jejunum did not exhibit adherence properties or express CD14, a receptor for the lipopolysaccharide-binding protein. The purification of large numbers of lamina propria macrophages by this procedure offers the opportunity to define the role of this cell in the physiological inflammation characteristic of normal intestinal mucosa and the pathological inflammation associated with small intestinal diseases.
黏膜巨噬细胞在小肠免疫事件调节和炎症反应中发挥着重要作用。由于目前尚无关于正常小肠巨噬细胞的信息,我们开发了一种分离和纯化空肠固有层巨噬细胞的技术,以研究其表型和活性。从正常人空肠切片中,通过中性蛋白酶消化分离出固有层单核细胞,然后进行逆流离心淘析以纯化巨噬细胞。通过该方法分离的细胞中CD3 +淋巴细胞含量<1%,并呈现出单核吞噬细胞的大小分布、形态特征、超微结构和吞噬活性。然而,与血液单核细胞不同,空肠黏膜巨噬细胞不表现出黏附特性,也不表达脂多糖结合蛋白的受体CD14。通过该方法大量纯化固有层巨噬细胞,为确定该细胞在正常肠黏膜生理性炎症以及与小肠疾病相关的病理性炎症中的作用提供了机会。