Low concentrations of recombinant granulocyte macrophage-colony stimulating factor derived from Chinese hamster ovary cells augments long-term bioactivity with delayed clearance in vitro
Low concentrations of recombinant granulocyte macrophage-colony stimulating factor derived from Chinese hamster ovary cells augments long-term bioactivity with delayed clearance in vitro
复制标题
源自中国仓鼠卵巢细胞的低浓度重组粒细胞巨噬细胞集落刺激因子可增强长期生物活性并延迟体外清除
DOI：
10.1016/j.cyto.2014.03.009
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发表时间：
2014
期刊：
影响因子：
3.8
通讯作者：
Nakata K.
中科院分区：
文献类型：
--
作者：
Hashimoto A;Tanaka T;Itoh Y;Yamagata A;Kitamura N;Tazawa R;Nakagaki K;Nakata K.
迄今为止，粒细胞巨噬细胞集落刺激因子（GM-CSF）的生物活性主要通过大肠杆菌或酵母细胞衍生的重组人GM-CSF（分别为erhGM-CSF和yrhGM-CSF）进行研究。然而，中国仓鼠卵巢细胞衍生的重组人GM-CSF （crhGM-CSF）和天然人GM-CSF是一种独特的分子，包括复杂的低聚糖修饰。在本研究中，我们将crhGM-CSF与erhGM-CSF和yrhGM-CSF进行了比较，重新评价了crhGM-CSF的生物活性。短期刺激（0.5 h）对中性粒细胞/单核细胞或外周血单核细胞（PBMCs）的激活作用低于低浓度（60 pM以下）的erhGM-CSF或yrhGM-CSF。不同rhgm - csf中期刺激（24 h）对tgf -1细胞、gm - csf依赖性细胞系或PBMCs的激活影响无显著差异。相比之下，低浓度的crhGM-CSF （15-30 pM）长期刺激（72-168 h）后，tgf -1细胞或PBMCs的增殖/存活率高于其他gm - csf处理的细胞。与其他rhgm - csf孵育72h后，tnf -1细胞的凋亡比例低于其他rhgm - csf。这些作用被crhGM-CSF的去盐化作用减弱。与erhGM-CSF或yrhGM-CSF相比，TF-1细胞和pbmc对crhGM-CSF的清除延迟，但不脱盐-crhGM-CSF。这些结果表明，低聚糖部分的唾液化延迟了GM-CSF的清除，从而引发了体外长期生物活性的增加。
To date, the biological activity of granulocyte macrophage-colony stimulating factor (GM-CSF) has been investigated by using mostlyEscherichia coli-or yeast cell-derived recombinant human GM-CSF (erhGM-CSF and yrhGM-CSF, respectively). However, Chinese hamster ovary cell-derived recombinant human GM-CSF (crhGM-CSF), as well as natural human GM-CSF, is a distinct molecule that includes modifications by complicated oligosaccharide moieties. In the present study, we reevaluated the bioactivity of crhGM-CSF by comparing it with those of erhGM-CSF and yrhGM-CSF. The effect of short-term stimulation (0.5 h) on the activation of neutrophils/monocytes or peripheral blood mononuclear cells (PBMCs) by crhGM-CSF was lower than those with erhGM-CSF or yrhGM-CSF at low concentrations (under 60 pM). Intermediate-term stimulation (24 h) among the different rhGM-CSFs with respect to its effect on the activation of TF-1 cells, a GM-CSF-dependent cell line, or PBMCs was not significantly different. In contrast, the proliferation/survival of TF-1 cells or PBMCs after long-term stimulation (72–168 h) was higher at low concentrations of crhGM-CSF (15–30 pM) than that of cells treated with other GM-CSFs. The proportion of apoptotic TF-1 cells after incubation with crhGM-CSF for 72 h was lower than that of cells incubated with other rhGM-CSFs. These effects were attenuated by desialylation of crhGM-CSF. Clearance of crhGM-CSF but not desialylated-crhGM-CSF by both TF-1 cells and PBMCs was delayed compared with that of erhGM-CSF or yrhGM-CSF. These results suggest that sialylation of oligosaccharide moieties delayed the clearance of GM-CSF, thus eliciting increased long-term bioactivityin vitro.