Brulez H F, ter Wee P M, Snijders S V, Donker A J, Verbrugh H A
Department of Nephrology, Academisch Ziekenhuis Vrije Universiteit Amsterdam, The Netherlands.
J Clin Pathol. 1999 Dec;52(12):901-9. doi: 10.1136/jcp.52.12.901.
Previous studies showed that the currently used dextrose based peritoneal dialysis fluids impair several leucocyte functions.
To determine which in vitro mononuclear leucocyte (monocyte) function tests most clearly reflect the biocompatibility of peritoneal dialysis fluid.
Monocytes were tested for phagocytic capacity, bactericidal activity, Fc and C3 receptor expression, and chemiluminescence response, and by analysis of the release of interleukin 8 (IL-8) and tumour necrosis factor alpha (TNF alpha) in the presence of test fluids. Cytokine release was studied in an alternative dynamic in vitro peritoneal dialysis model in which monocytes were exposed to test fluid that was continuously equilibrated with an interstitial fluid-like medium through a microporous membrane. The chemiluminescence response by stressed monocytes was also tested after an 18 h recovery period. All tests were performed during or after exposure to different degrees of glycerol induced osmotic stress and after exposure to a 1% milk-whey derived, polypeptide enriched test fluid. Cells incubated in 0.1% gel Hanks buffer (GH) served as control.
Osmotic stress induced impairment of leucocyte function was found by the chemiluminescence assay (mean (SEM): 179 (20)% v 138 (23)% after 30 minutes in 0.5% and 1.5% glycerol, respectively) and by the analysis of IL-8 released by monocytes (44 (9) ng in 0.7% glycerol v 40 (7) ng in 2.0% glycerol). Only the chemiluminescence assay showed a protective effect of polypeptides on leucocyte function (after > or = 60 minutes). If monocytes were allowed to recover in culture medium after exposure to test fluids, the changes in chemiluminescence response appeared to be reversible after a 30 minute exposure, but became more pronounced after 60 and 120 minutes. The phagocytosis and bacterial killing assays were less sensitive. The observations carried out with the phagocytosis assay did not correspond with the Fc or C3 receptor density data.
The release of IL-8 by peripheral blood monocytes in a two compartment model and their chemiluminescence response are appropriate assays for the assessment of changes in leucocyte function in response to different peritoneal dialysis fluids.
先前的研究表明，目前使用的基于葡萄糖的腹膜透析液会损害多种白细胞功能。
确定哪种体外单核白细胞（单核细胞）功能测试能最清晰地反映腹膜透析液的生物相容性。
对单核细胞进行吞噬能力、杀菌活性、Fc和C3受体表达、化学发光反应测试，并分析在测试液存在下白细胞介素8（IL - 8）和肿瘤坏死因子α（TNFα）的释放情况。细胞因子释放是在一种替代性动态体外腹膜透析模型中进行研究的，在该模型中，单核细胞暴露于通过微孔膜与类似组织间液的介质持续平衡的测试液中。在18小时恢复期后，还测试了应激单核细胞的化学发光反应。所有测试均在暴露于不同程度的甘油诱导的渗透应激期间或之后，以及暴露于1%乳清来源的富含多肽的测试液之后进行。在0.1%凝胶汉克斯缓冲液（GH）中孵育的细胞用作对照。
通过化学发光测定法（平均值（标准误）：在0.5%和1.5%甘油中分别孵育30分钟后，分别为179（20）%和138（23）%）以及单核细胞释放的IL - 8分析（在0.7%甘油中为44（9）纳克，在2.0%甘油中为40（7）纳克）发现渗透应激会诱导白细胞功能受损。只有化学发光测定法显示多肽对白细胞功能有保护作用（在≥60分钟后）。如果单核细胞在暴露于测试液后在培养基中恢复，化学发光反应的变化在暴露30分钟后似乎是可逆的，但在60分钟和120分钟后变得更加明显。吞噬作用和细菌杀伤测定法不太敏感。吞噬作用测定法的观察结果与Fc或C3受体密度数据不相符。
在双室模型中，外周血单核细胞释放IL - 8及其化学发光反应是评估不同腹膜透析液引起的白细胞功能变化的合适测定方法。
