Jaswon M S, Jones H M, Linch D C
Department of Haematology, University College, London, United Kingdom.
Pediatr Res. 1994 Nov;36(5):623-7. doi: 10.1203/00006450-199411000-00016.
To investigate further the susceptibility to infection of newborn infants, particularly those born prematurely, we have used a "whole blood" flow cytometric assay to compare the respiratory burst activity in recombinant human granulocyte-macrophage colony stimulating factor "primed" neutrophils obtained from healthy adults, term infants, and preterm newborn infants. The use of whole blood avoids prior cell separation procedures that may cause artifactual activation or priming. In healthy adults (n = 21), the bacterial cell wall peptide N-formyl-methionyl-leucyl-phenylalanine induced little neutrophil respiratory burst activity, suggesting that the circulating cell is relatively quiescent. Prior exposure to recombinant human granulocyte-macrophage colony stimulating factor augmented the median N-formyl-methionyl-leucyl-phenylalanine response by 425%. In cord blood from full-term neonates (n = 9), recombinant human granulocyte-macrophage colony stimulating factor produced less enhancement of the N-formyl-methionyl-leucyl-phenylalanine response (345%), but the absolute level of respiratory burst activity was at least as great as in adults, suggesting that the neutrophils are fully functional and partially primed after delivery. In preterm infants receiving intensive care (n = 10), the degree of priming was similar to that in neutrophils from term infants (344%), although the absolute level of respiratory burst activity was reduced (p = 0.0003). In response to stimulation with phorbol ester, 73.5% (18-99%) (median and range) of neutrophils obtained from adults and 77.6% (50-92%) from term babies exhibit respiratory burst activity detectable in the whole blood assay. However, in neutrophils obtained from preterm infants, there was a significant reduction in the phorbol ester-induced respiratory burst, with only 32.9% (21-61%) of cells responding (p = 0.0129).
为了进一步研究新生儿尤其是早产儿的感染易感性,我们采用了一种“全血”流式细胞术检测方法,比较从健康成人、足月儿和早产新生儿获取的经重组人粒细胞巨噬细胞集落刺激因子“预刺激”的中性粒细胞的呼吸爆发活性。使用全血避免了可能导致人为激活或预刺激的先前细胞分离程序。在健康成人(n = 21)中,细菌细胞壁肽N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸诱导的中性粒细胞呼吸爆发活性很低,这表明循环中的细胞相对静止。预先暴露于重组人粒细胞巨噬细胞集落刺激因子可使N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸反应的中位数增加425%。在足月新生儿的脐血(n = 9)中,重组人粒细胞巨噬细胞集落刺激因子对N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸反应的增强作用较小(345%),但呼吸爆发活性的绝对水平至少与成人相同,这表明中性粒细胞在出生后功能完全且部分处于预刺激状态。在接受重症监护的早产儿(n = 10)中,预刺激程度与足月儿中性粒细胞相似(344%),尽管呼吸爆发活性的绝对水平降低了(p = 0.0003)。在用佛波酯刺激时,从成人获取的中性粒细胞中有73.5%(18 - 99%)(中位数和范围)以及从足月儿获取的中性粒细胞中有77.6%(50 - 92%)在全血检测中表现出可检测到的呼吸爆发活性。然而,在从早产儿获取的中性粒细胞中,佛波酯诱导的呼吸爆发显著降低,只有32.9%(21 - 61%)的细胞有反应(p = 0.0129)。