Cytoplasts made from human blood polymorphonuclear leukocytes with or without heat: preservation of both motile function and respiratory burst oxidase activity.
Cytoplasts made from human blood polymorphonuclear leukocytes with or without heat: preservation of both motile function and respiratory burst oxidase activity.
复制标题
由人血液多形核白细胞制成的细胞质，加热或不加热：保持运动功能和呼吸爆发氧化酶活性。
DOI：
10.1073/pnas.84.2.454
复制
发表时间：
1987
影响因子：
11.1
通讯作者：
VanBlaricom,G
中科院分区：
文献类型：
--
作者：
Malawista,SE;VanBlaricom,G
多形核白细胞（PMN）的无核片段（细胞质）是一种简化的系统，可用于阐明细胞功能改变的特定途径。目前使用的PMN细胞质在可激活的呼吸爆发氧化酶活性或运动功能方面存在缺陷。通过在没有细胞松弛素B的Ficoll的不连续梯度上离心PMN，我们创造了颗粒贫乏的细胞质，其中这两种能力都得到了保存。具体来说，在适当的刺激下，它们产生超氧阴离子（O2-）并还原硝基蓝四氮唑染料；它们对激光微照射破坏的红细胞或对特定的化学引诱剂（10 nM）和C5a（酶酶活性血清）有趋化反应；它们吞下并杀死葡萄球菌。我们可以通过将悬浮细胞预热（45℃）增加时间来逐步提高这些片段的产量，但这些处理伴随着具有可活化氧化酶活性的细胞质百分比的减少，并且细胞质逐渐无法摄取和杀死葡萄球菌。这些容易制造的多能细胞质很容易用于PMN生理学的研究。
Anucleate fragments (cytoplasts) from polymorphonuclear leukocytes (PMN) are simplified systems that can be used to elucidate specific pathways by which cell function is altered. PMN cytoplasts in current use are defective either in activatable respiratory burst oxidase activity or in motile function. By centrifugation of PMN on discontinuous gradients of Ficoll without cytochalasin B, we have created granule-poor cytoplasts in which both these capacities are preserved. Specifically, they generate superoxide anion (O2-.) and reduce nitroblue tetrazolium dye on appropriate stimulation; they respond chemotactically to erythrocytes destroyed by laser microirradiation or to the specific chemoattractants fMet-Leu-Phe (10 nM) and C5a (zymosan-activated serum); and they ingest and kill staphylococci. We can improve the yield of these fragments progressively by preheating (45 degrees C) the cells in suspension for increasing periods of time, but those treatments are attended by a decreasing percentage of cytoplasts with activatable oxidase activity, and a progressive inability of the cytoplasts to ingest and to kill staphylococci. These easily made and multipotent cytoplasts readily lend themselves to studies of PMN physiology.