Shirafuji N, Asano S, Matsuda S, Watari K, Takaku F, Nagata S
Department of Hematology-Oncology, University of Tokyo, Japan.
Exp Hematol. 1989 Feb;17(2):116-9.
[3H]thymidine uptake by NFS-60 cells in microcultures was found to increase in a linear fashion with the increasing doses of purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). Such increases were found neither with rhG-CSF samples pretreated with rabbit anti-rhG-CSF serum nor with other human colony-stimulating factors such as granulocyte-macrophage colony-stimulating factor (hGM-CSF) or macrophage colony-stimulating factor (hM-CSF). Based on these findings, sera from normal persons and patients with severe infections or various hematological disorders were tested after dialysis using this system in order to determine whether G-CSF levels in sera can be estimated or not. In ten normal persons, five patients with acute myelogenous leukemia (AML M1, M2, and M3), five with myelodysplastic syndrome, and four with chronic myelogenous leukemia, no increases in [3H]thymidine uptake were found within the dose range of 0.4 microliters to 50 microliters. In contrast, linear dose responses parallel to a G-CSF standard curve were observed in one patient with a severe bacterial infection, four with aplastic anemia, two with acute myelomonocytic leukemia (AMMoL) (M4), and two with idiopathic neutropenia tested. From the standard curve, the probable levels of G-CSF were calculated as follows: approximately 200 pg/ml with infection, 130-220 pg/ml with aplastic anemia, 150 and 200 pg/ml with AMMoL, and 1120 and 1200 pg/ml with idiopathic neutropenia. The activities of sera were reduced by the anti-rhG-CSF serum pretreatment in the same way as documented in the case of rhG-CSF. Furthermore, the level in a patient with a severe infection became undetectable soon after elimination of the infection and blood neutrophil counts had returned to normal. These findings indicate that the microbioassay system will be useful for measuring circulating G-CSF levels which would fluctuate in accord with requirements for stimulating neutrophil production or with abnormal production of hG-CSF.
研究发现，在微量培养中，NFS - 60细胞对[3H]胸腺嘧啶核苷的摄取量随纯化重组人粒细胞集落刺激因子（rhG - CSF）剂量的增加呈线性增加。用兔抗rhG - CSF血清预处理的rhG - CSF样品以及其他人类集落刺激因子，如粒细胞 - 巨噬细胞集落刺激因子（hGM - CSF）或巨噬细胞集落刺激因子（hM - CSF），均未出现这种增加。基于这些发现，使用该系统对正常人和严重感染或各种血液系统疾病患者的血清进行透析后检测，以确定血清中G - CSF水平是否可被估算。在10名正常人、5名急性髓性白血病患者（AML M1、M2和M3）、5名骨髓增生异常综合征患者以及4名慢性髓性白血病患者中，在0.4微升至50微升的剂量范围内，未发现[3H]胸腺嘧啶核苷摄取量增加。相反，在1名严重细菌感染患者、4名再生障碍性贫血患者、2名急性粒单核细胞白血病（AMMoL）（M4）患者以及2名特发性中性粒细胞减少症患者中，观察到与G - CSF标准曲线平行的线性剂量反应。根据标准曲线，计算出的G - CSF可能水平如下：感染患者约为200 pg/ml，再生障碍性贫血患者为130 - 220 pg/ml，AMMoL患者为150和200 pg/ml，特发性中性粒细胞减少症患者为1120和1200 pg/ml。血清活性经抗rhG - CSF血清预处理后降低，与rhG - CSF情况相同。此外，1名严重感染患者在感染消除且血液中性粒细胞计数恢复正常后不久，其体内水平变得无法检测。这些发现表明，该微生物测定系统将有助于测量循环G - CSF水平，其会根据刺激中性粒细胞生成的需求或hG - CSF的异常生成而波动。
