Gilbert C S, Parmley R T, Rice W G, Kinkade J M
Department of Pediatrics, University of Texas Health Science Center, San Antonio 78284-7810.
J Histochem Cytochem. 1993 Jun;41(6):837-49. doi: 10.1177/41.6.8315276.
Studies have demonstrated significant heterogeneity in neutrophil granule morphology and physical density. This study evaluated the heterogeneity morphometrically, morphologically, cytochemically, and biochemically. Intact human peripheral blood neutrophils collected from normal volunteers and a patient with Chédiak-Higashi syndrome (CHS) and isolated normal neutrophil granules were processed for ultrastructural morphology and peroxidase staining. Intact cells, nuclei, and granule profiles were analyzed by computer-assisted planimetry. Peroxidase-positive granules (PPG) represented about 40% of normal neutrophil granules and covered the entire spectrum of granule size. PPG in the least-dense fractions of isolated granules were significantly smaller than in higher-density fractions. PPG in low- and intermediate-density fractions differed from high-density fraction by moderate to strong vicinal glycol staining with Thiéry's periodate-thiocarbohydrazide-silver proteinate method. Differing ratios of % beta-glucuronidase/% myeloperoxidase (MPO) across granule fractions indicated PPG heterogeneity. Morphometric analysis of neutrophils treated with 1 microM calcium ionophore A23187 did not show significant differences in PPG size or number. Biochemically analyzed MPO in these cells was preserved, although the number of peroxidase-negative granules (PNG) and levels of vitamin B12-binding protein were markedly decreased. In CHS, about 20% of granules were PPG. Analysis of CHS neutrophils revealed the persistence of microgranules similar to normals. PNG number and volume fractions of PPG and TG were not different from normals. Complex heterogeneity of normal PPG was quantitated using morphometry and appeared preserved in ionophore-treated cells but was uniquely modified in CHS.
研究表明中性粒细胞颗粒形态和物理密度存在显著异质性。本研究从形态计量学、形态学、细胞化学和生物化学方面评估了这种异质性。采集正常志愿者和一名患有切-东综合征（CHS）患者的完整人外周血中性粒细胞，以及分离出的正常中性粒细胞颗粒，进行超微结构形态学和过氧化物酶染色处理。通过计算机辅助平面测量法分析完整细胞、细胞核和颗粒轮廓。过氧化物酶阳性颗粒（PPG）约占正常中性粒细胞颗粒的40%，涵盖了颗粒大小的整个范围。分离颗粒中密度最低部分的PPG明显小于高密度部分的PPG。用蒂埃里高碘酸盐-硫代碳酰肼-银蛋白法对低密度和中密度部分的PPG进行中度至强邻二醇染色，结果显示其与高密度部分不同。不同颗粒部分的β-葡萄糖醛酸酶/髓过氧化物酶（MPO）百分比比值不同，表明PPG存在异质性。用1 microM钙离子载体A23187处理的中性粒细胞的形态计量学分析显示，PPG大小或数量没有显著差异。尽管过氧化物酶阴性颗粒（PNG）数量和维生素B12结合蛋白水平明显降低，但这些细胞中生物化学分析的MPO得以保留。在CHS中，约20%的颗粒为PPG。对CHS中性粒细胞的分析显示，与正常情况类似，微颗粒持续存在。PPG和TG的PNG数量和体积分数与正常情况无异。使用形态计量学对正常PPG的复杂异质性进行了定量，结果显示其在离子载体处理的细胞中得以保留，但在CHS中发生了独特的改变。
