淋巴因子刺激的巨噬细胞对荧光微球的吞噬作用：一种快速的新检测方法。

Lymphokine-stimulated macrophage phagocytosis of fluorescent microspheres: a rapid new assay.

作者信息

Dunn P A, Eaton W R, Lopatin E D, McEntire J E, Papermaster B W

出版信息

J Immunol Methods. 1983 Nov 11;64(1-2):71-83. doi: 10.1016/0022-1759(83)90385-x.

Abstract

A highly sensitive and rapid in vitro macrophage phagocytosis assay is described for screening of lymphokine preparations with macrophage activating properties. Non-induced mouse peritoneal macrophages were incubated on glass slides for 60 min in the presence of fluorescent 2 micron latex beads and partially purified lymphokine fractions or media control. Lymphokine samples were prepared from culture supernatants of the B lymphoblastoid cell line RPMI 1788 by high performance liquid chromatography of a soluble trichloroacetic acid extract of concentrated culture supernatant. Phagocytosis was measured by direct counts of the percent phagocytic cells and the number of intracellular beads per 100 cells as seen by fluorescence microscopy. Phagocytic uptake in the presence of as little as 0.1 ng of active lymphokine could readily be determined qualitatively by correlation plots and quantitatively by appropriate statistical programs for data processing by computer. This technique provides a rapid, reproducible, screening assay for biological activity of macrophage activating lymphokines and other compounds affecting macrophage phagocytosis.

摘要

本文描述了一种高度灵敏且快速的体外巨噬细胞吞噬试验，用于筛选具有巨噬细胞激活特性的淋巴因子制剂。将未诱导的小鼠腹腔巨噬细胞与荧光2微米乳胶珠以及部分纯化的淋巴因子组分或培养基对照一起在载玻片上孵育60分钟。淋巴因子样品通过对浓缩培养上清液的可溶性三氯乙酸提取物进行高效液相色谱法，从B淋巴母细胞系RPMI 1788的培养上清液中制备。通过荧光显微镜观察，直接计数吞噬细胞百分比和每100个细胞内珠子的数量来测量吞噬作用。在存在低至0.1 ng活性淋巴因子的情况下，吞噬摄取可通过相关图定性地轻松确定，并通过适当的计算机数据处理统计程序进行定量测定。该技术为巨噬细胞激活淋巴因子和其他影响巨噬细胞吞噬作用的化合物的生物活性提供了一种快速、可重复的筛选试验。