Appel S H, Wellhausen S R, Montgomery R, DeWeese R C, Polk H C
Price Institute of Surgical Research, Department of Surgery, University of Louisville School of Medicine, Kentucky 40292.
J Surg Res. 1989 Jul;47(1):39-44. doi: 10.1016/0022-4804(89)90045-0.
For much of the last decade, an increasing number of surgeons have been interested in objective assessment of cellular contributors to host defense function. In order to study many of these processes, it is apparently desirable that the cells be isolated to the extent feasible for the purpose of analyzing a more or less pure population of cellular elements. The purpose of this paper is to describe the physiologic activation of mononuclear cells that occurs as a result of the isolation process. Therefore, it follows logically that such cells are therein intrinsically less responsive to further physiologic manipulation in vitro. Analyses of such data without an awareness of this intrinsic aberration will undoubtedly lead to misinterpretation of the capacity of such cells for further modulation by immunostimulants or by the intrinsic processes related to injury, anesthesia, and operation. Furthermore, it may indicate that certain agents, e.g., cytokines, are unable to stimulate cellular function when, in fact, the defense function of the cell has been initially stimulated by the isolation procedure. Fractionation of human peripheral blood over Hypaque-Ficoll and subsequent purification of monocytes by adherence to plastic lead to an increase in the relative density of HLA-DR on monocytes. This increase occurred when carried out in endotoxin lipopolysaccharide (LPS)-contaminated or LPS-depleted reagents. LPS, added experimentally to whole blood, enhanced HLA-DR expression on monocytes without further manipulation. Monocyte HLA-DR expression measured in whole blood was reduced in patients with major sepsis (n = 19) compared to normal subjects (n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)
在过去十年的大部分时间里,越来越多的外科医生对客观评估参与宿主防御功能的细胞成分感兴趣。为了研究其中许多过程,显然希望在可行的范围内分离细胞,以便分析或多或少纯净的细胞成分群体。本文的目的是描述由于分离过程而发生的单核细胞的生理激活。因此,合乎逻辑的是,这些细胞在体外对进一步的生理操作的反应性内在地较低。如果没有意识到这种内在异常而对这些数据进行分析,无疑会导致对这些细胞通过免疫刺激剂或与损伤、麻醉和手术相关的内在过程进行进一步调节的能力的错误解读。此外,这可能表明某些因子,如细胞因子,无法刺激细胞功能,而实际上细胞的防御功能最初已被分离程序刺激。通过在淋巴细胞分离液上分离人外周血并随后通过贴壁于塑料培养皿纯化单核细胞,会导致单核细胞上HLA-DR的相对密度增加。在内毒素脂多糖(LPS)污染或不含LPS的试剂中进行此操作时,都会出现这种增加。实验性地向全血中添加LPS,无需进一步操作即可增强单核细胞上HLA-DR的表达。与正常受试者(n = 10)相比,严重脓毒症患者(n = 19)全血中测得的单核细胞HLA-DR表达降低。(摘要截短于250字)