In vitro evaluation of the effect of a novel immunosuppressive agent, FTY720, on the function of feline neutrophils
In vitro evaluation of the effect of a novel immunosuppressive agent, FTY720, on the function of feline neutrophils
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DOI：
10.2460/ajvr.67.4.588
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发表时间：
2006-04-01
影响因子：
1
通讯作者：
Gregory, CR
中科院分区：
文献类型：
--
作者：
Chen, YJ;Kyles, AE;Gregory, CR
目的：通过体外实验评估新型免疫抑制剂FTY720对猫外周中性粒细胞生物学功能（迁移、吞噬和活性氧[ROS]产生）的影响，并确定FTY720对猫外周中性粒细胞的细胞毒性作用。样本群- 8只健康猫外周血中性粒细胞。操作步骤：从8只猫的血液样本中分离外周中性粒细胞，并暴露于磷酸化形式的FTY720 （FTY720- p）。进行了基于荧光的体外迁移评估。采用双色流式细胞仪检测微生物的吞噬作用和活性氧的产生。从猫身上获得的全血样本与不同浓度的FTY720-P荧光素标记的金黄色葡萄球菌和二氢乙啶一起孵育。用碘化丙啶染色评价细胞毒性作用。结果：FTY720-P的加入对猫外周中性粒细胞的吞噬和ROS的产生有轻微的不显著的降低。添加FTY720-P后，猫外周中性粒细胞的迁移活性显著增加。添加临床使用浓度的FTY720-P不会增加猫外周中性粒细胞的死亡率。结论和临床意义：fty720在体外不抑制猫外周中性粒细胞的关键功能。
Objective-To use in vitro assays to evaluate the effects of a novel immunosuppressive agent, FTY720, on biological functions (migration, phagocytosis, and production of reactive-oxygen species [ROS]) of feline peripheral neutrophils and determine the cytotoxic effects of FTY720 on feline peripheral neutrophils.Sample Population-Peripheral neutrophils obtained from 8 healthy cats.Procedure-Peripheral neutrophils were isolated from blood samples obtained from the 8 cats and exposed to the phosphorylated form of FTY720 (FTY720-P). A fluorescence-based in vitro evaluation of migration was performed. Phagocytosis of microbes and production of ROS were evaluated by use of a 2-color flow cytometry system. Samples of whole blood obtained from the cats were incubated with various concentrations of FTY720-P fluorescein-labeled Staphylococcus aureus, and dihydroethidium. Cytotoxic effects were evaluated by use of propidium iodide staining.Results-Addition of FTY720-P caused a slight nonsignificant decrease in phagocytosis and production of ROS by feline peripheral neutrophils. Migration activity of feline peripheral neutrophils was significantly increased by the addition of FTY720-P Addition of FTY720-P at concentrations considered for clinical use did not increase the death rate of feline peripheral neutrophils.Conclusions and Clinical Relevance-FTY720 does not inhibit critical functions of feline peripheral neutrophils in vitro.