Lionetti F J, Lin P S, Mattaliano R J, Hunt S M, Valeri C R
Exp Hematol. 1980 Mar;8(3):304-17.
Scanning microscopic and functional studies were made of granulocytes isolated from CPD anticoagulated whole blood by counterflow centrifugation in a Beckman JE-6 rotor. The collection buffer was phosphate (20 mM) buffered saline (280 mOsM) with glucose (29 mM) and human serum albumin (1.2% w/v). The final suspension contained less than 2% mononuclear cells and 5% red cells. Incubation and fixation at various temperatures revealed two distinct temperature dependent shape transformations. At 22, 37, 40 and 45 degrees C granulocytes were ameboid with extensive highly textured veils. These smoothed progressively, bullae and blebs formed, and membranes peeled finally leaving nonfunctional spheres with smooth surfaces. At 4 degrees C, granulocytes were irregular spheres, less rugose but with numerous microvilli and nodules. Veiling was absent. Phagocytosis, initially low, progressively declined over 48 h while cell surfaces become smooth. Some formed blebs, but all terminated as nonfunctional spheres with untextured surfaces containing occasional large single holes. Cellular stability estimated from changes in volume distributions, and phagocytosis by microfluorescence measurements of yeast and latex particle ingestion were also temperature dependent and paralleled the shape progressions. It is concluded that at body (37 degrees C) or fever (40 degrees C) temperatures, granulocytes have dynamic membrane surfaces characterized by extensive veiling and high function. At 4 degrees C they are relatively inactive spheres devoid of pseudopodia or veils, yet functional at slow rates.
采用贝克曼JE - 6转头通过逆流离心法对从CPD抗凝全血中分离出的粒细胞进行了扫描显微镜和功能研究。收集缓冲液为含葡萄糖（29 mM）和人血清白蛋白（1.2% w/v）的磷酸盐（20 mM）缓冲盐水（280 mOsM）。最终悬浮液中单核细胞少于2%，红细胞少于5%。在不同温度下进行孵育和固定揭示了两种不同的温度依赖性形态转变。在22、37、40和45摄氏度时，粒细胞呈阿米巴样，有广泛的高度纹理化的膜。这些膜逐渐平滑，形成大泡和小泡，最后细胞膜剥离，留下表面光滑的无功能球体。在4摄氏度时，粒细胞为不规则球体，皱纹较少但有许多微绒毛和结节。没有膜形成。吞噬作用最初较低，在48小时内逐渐下降，同时细胞表面变得光滑。一些细胞形成了小泡，但最终都变成了表面无纹理、偶尔有大的单孔的无功能球体。根据体积分布变化估计的细胞稳定性以及通过微量荧光测量酵母和乳胶颗粒摄取来评估的吞噬作用也与温度有关，并与形态变化平行。结论是，在体温（37摄氏度）或发热（40摄氏度）时，粒细胞具有以广泛的膜形成和高功能为特征的动态膜表面。在4摄氏度时，它们是相对不活跃的球体，没有伪足或膜，但功能速率较慢。
