Pryzwansky K B, Breton-Gorius J
Lab Invest. 1985 Dec;53(6):664-71.
Two main classes of primary granules have been identified in human neutrophils by peroxidase cytochemistry and electron microscopy, spherical granules and elongated granules, containing crystals with well-defined periodicity. In the present study, we report another subpopulation of primary granules, distinguishable from the other primary granules by their small size, strong peroxidase reactivity under selective incubation conditions with diaminobenzidine, stage of neutrophil maturation at which they appear, and distinctive morphological characteristics. Early promyelocytes, reacted with diaminobenzidine before fixation, demonstrate no reactivity in the large primary granules, but strong peroxidase activity in the endoplasmic reticulum and Golgi apparatus. At the end of the promyelocyte stage of maturation, when the endoplasmic reticulum decreases in peroxidase reactivity, small, round or elongated granules (100 to 200 nm) are observed, reacting strongly for peroxidase. These small granules persist during maturation. The data suggest that these small granules are packaged after the large primary granules and before the secondary granules. All three classes of primary granules exhibit peroxidase activity when fixed prior to incubation in diaminobenzidine at neutral or alkaline pH. Distinctive morphological characteristics of the small primary granules are observed in circulating neutrophils. The small granules are arranged in chains or clusters primarily at the cell margin or uropod in moving cells and are aligned along the axis of cell polarity. This granule association is more evident in adherent neutrophils, particularly after short incubation with phorbol myristate acetate. Chains consisting of as many as 17 small granules are observed. Stereo high voltage electron microscopy of whole-mount preparations of adherent neutrophils reveals chains of small granules apparently interconnected by microtrabeculae. Thus, the small granules observed in thin sections do not represent a transverse section of elongated granules, and the interconnection of granules by microtrabeculae may determine the arrangement of granules. These data support the existence of subpopulations of primary granules which contain distinct forms of myeloperoxidase.
通过过氧化物酶细胞化学和电子显微镜技术，在人类中性粒细胞中已鉴定出两类主要的初级颗粒，即球形颗粒和细长颗粒，后者含有具有明确周期性的晶体。在本研究中，我们报告了另一类初级颗粒亚群，它们与其他初级颗粒的区别在于体积小、在与二氨基联苯胺的选择性孵育条件下具有较强的过氧化物酶反应性、出现时中性粒细胞的成熟阶段以及独特的形态特征。早期早幼粒细胞在固定前与二氨基联苯胺反应，在大的初级颗粒中未显示反应性，但在内质网和高尔基体中具有较强的过氧化物酶活性。在早幼粒细胞成熟阶段结束时，当内质网的过氧化物酶反应性降低时，观察到小的圆形或细长颗粒（100至200纳米），对过氧化物酶反应强烈。这些小颗粒在成熟过程中持续存在。数据表明，这些小颗粒是在大的初级颗粒之后、次级颗粒之前包装形成的。当在中性或碱性pH下于二氨基联苯胺中孵育前固定时，所有三类初级颗粒均表现出过氧化物酶活性。在循环中的中性粒细胞中观察到小初级颗粒的独特形态特征。小颗粒主要以链状或簇状排列在移动细胞的细胞边缘或尾足处，并沿细胞极性轴排列。这种颗粒关联在贴壁中性粒细胞中更明显，尤其是在用佛波酯肉豆蔻酸酯短暂孵育后。观察到由多达17个小颗粒组成的链。对贴壁中性粒细胞整装标本进行立体高压电子显微镜检查发现，小颗粒链显然通过微梁相互连接。因此，在薄片中观察到的小颗粒并不代表细长颗粒的横切面，并且颗粒通过微梁的相互连接可能决定了颗粒的排列。这些数据支持存在含有不同形式髓过氧化物酶的初级颗粒亚群。
