Wolach B, Ben Dor M, Chomsky O, Gavrieli R, Shinitzky M
Department of Pediatrics, Meir General Hospital, Tel-Aviv University, Israel.
J Leukoc Biol. 1992 Apr;51(4):324-8. doi: 10.1002/jlb.51.4.324.
Membrane lipid fluidity of peripheral blood polymorphonuclear cells (PMNs) of 24 newborn infants, 2-4 days after birth, was determined by steady-state fluorescence polarization with 1,6-diphenyl 1,3,5-hexatriene (DPH) as a probe and compared with that of PMNs from 23 adults. Measurements with intact cells, which correspond to all cellular lipid domains, did not display any statistically significant difference between PMNs of the two groups. However, application of bixinoyl glucosamine, a membrane-impermeable fluorescence quencher, revealed that the PMN plasma membrane of the newborn is about 23% more fluid than that of the adult. Total cholesterol-to-phospholipid ratio of newborn PMNs was found to be lower by about 10% than that of the adult, which could account for the difference in their plasma membrane fluidity. The possible implication of this finding for the deficit in chemotactic ability of leukocytes from newborns was tested with neonatal PMNs that have incorporated cholesteryl hemisuccinate (CHS), an efficient plasma membrane rigidifier. In all neonatal PMNs tested a mild incorporation of CHS (0.5-1 min incubation in 50 micrograms/ml dispersion) caused a significant improvement in their net chemotaxis, from an average value of 28 +/- 7 to 43 +/- 11. Longer incubations with CHS caused a gradual decrease in chemotactic ability that approached the basal level after about 5 min incubation. The net chemotaxis in adult PMNs was significantly higher than that of neonatal PMNs (72 +/- 13) and was gradually inhibited by incorporation of CHS without any initial augmentation. Based on these results it was estimated that about 27% of the chemotactic deficit of neonatal PMNs is mediated by their immature fluid membrane.
采用以1,6 - 二苯基 - 1,3,5 - 己三烯(DPH)为探针的稳态荧光偏振法,测定了24名出生后2 - 4天的新生儿外周血多形核白细胞(PMN)的膜脂流动性,并与23名成年人的PMN进行比较。对完整细胞(对应所有细胞脂质结构域)的测量结果显示,两组PMN之间没有任何统计学上的显著差异。然而,使用膜不透性荧光猝灭剂辣椒玉红素葡萄糖胺发现,新生儿的PMN质膜比成年人的质膜流动性约高23%。发现新生儿PMN的总胆固醇与磷脂比值比成年人低约10%,这可能是其质膜流动性差异的原因。用掺入了半琥珀酸胆固醇(CHS,一种有效的质膜硬化剂)的新生儿PMN测试了这一发现对新生儿白细胞趋化能力缺陷的可能影响。在所有测试的新生儿PMN中,轻度掺入CHS(在50微克/毫升分散液中孵育0.5 - 1分钟)会使其净趋化作用显著改善,从平均值28±7提高到43±11。与CHS孵育时间延长会导致趋化能力逐渐下降,孵育约5分钟后接近基础水平。成年人PMN的净趋化作用显著高于新生儿PMN(72±13),并且掺入CHS会使其逐渐受到抑制,且没有任何初始增强作用。基于这些结果,估计新生儿PMN约27%的趋化缺陷是由其不成熟的流动性膜介导的。