Anderson D C, Hughes B J, Smith C W
J Clin Invest. 1981 Oct;68(4):863-74. doi: 10.1172/jci110341.
To determine the mechanism(s) of diminished, stimulated, and directed migration of neonatal (N) polymorphonuclear leukocytes (PMN), chemotactic factor (CF) sensory and PMN effector functions were studied in healthy N and adult or maternal controls (C). N PMN demonstrated high affinity binding for N-formyl-methionyl-leucyl-[3H]phenylalanine (fMLP), which was saturable between 40 and 100 nM as observed with C PMN. The kinetics of binding and the characteristics of dissociation of binding by N PMN were equivalent to control PMN. Both "threshold" and "peak" concentrations (1 and 10 nM, respectively) of fMLP effected comparable PMN chemiluminescence among neonates and controls. An equivalent threshold concentration (0.05 nM) of fMLP effected N and C PMN shape change in suspension, and a maximally effective concentration (5 nM) induced comparable bipolar configuration, although uropod formation was only 38 +/- 8% of N PMN, compared with 73 +/- 11% of C PMN (P less than 0.01). Striking abnormalities of N PMN adherence were identified: mean +/- SD base-line (unstimulated) N adherence values (39 +/- 8%) were equal to C (38 +/- 9%), but diminished increments in response to single CF stimuli were noted among N (fMLP: 42 +/- 7% (N), 70 +/- 11% (C); C5a: 41 +/- 6% (N), 68 +/- 6% (C); BCF: 41 +/- 6% (N), 63 +/- 9% (C), P less than 0.01 for each CF). On sequential exposure to increasing concentrations of CF N PMN failed to demonstrate expected decreased adherence values; sequential stimuli with fMLP (0.1 nM, 10 nM) or C5a (8 microgram protein/ml, 32 microgram protein/ml) effected mean +/- 1 SD values of 51 +/- 9% (N), 30 +/- 9% (C), and 34 +/- 10 (N), 48 +/- 14% (C), respectively. As demonstrated with a latex bead-binding technique, N PMN failed to redistribute adhesion sites to the cell's tail under the same experimental conditions; in 21 N samples studied, restricted unipolar binding occurred in 33 +/- 8% (fMLP) or 37 +/- 7% (C5a) of PMN in contrast to C values of 70% (fMLP), or 71% (C5a), P less than 0.001. Similar findings were observed when PMN were preincubated with colchicine (25 microgram/ml); expected diminished adherence scores (compared with base-line values) were demonstrated with C PMN but not with N cells, P less than 0.01. Additionally colchicine-induced redistribution of adhesion sites as was observed with C samples (72 +/- 14% unipolar binding) was significantly (P less than 0.001) less among N PMN (31 +/- 11% unipolar binding). These investigations indicate that CF sensory mechanisms of N PMN are normal, compared with healthy adult or maternal controls. Diminished stimulated locomotion of the N PMN may be functionally related to reduced modulation of cell adhesiveness by chemotactic stimulation.
为了确定新生儿(N)多形核白细胞(PMN)趋化性降低、受刺激和定向迁移的机制,我们研究了健康新生儿及成人或母体对照(C)中趋化因子(CF)的感知功能和PMN效应功能。N PMN对N - 甲酰甲硫氨酰 - 亮氨酰 - [³H]苯丙氨酸(fMLP)表现出高亲和力结合,与C PMN一样,在40至100 nM之间可饱和。N PMN的结合动力学和解离特性与对照PMN相当。fMLP的“阈值”和“峰值”浓度(分别为1和10 nM)在新生儿和对照中引起相当的PMN化学发光。fMLP的等效阈值浓度(0.05 nM)使N和C PMN在悬浮液中发生形态变化,最大有效浓度(5 nM)诱导出相当的双极形态,尽管N PMN的尾足形成仅为C PMN的38±8%,而C PMN为73±11%(P<0.01)。发现N PMN的黏附存在显著异常:平均±标准差基线(未刺激)N黏附值(39±8%)与C(38±9%)相等,但在对单一CF刺激的反应中,N的增量减少(fMLP:42±7%(N),70±11%(C);C5a:41±6%(N),68±6%(C);BCF:41±6%(N),63±9%(C),每种CF的P均<0.01)。在依次暴露于浓度增加的CF时,N PMN未能表现出预期的黏附值降低;用fMLP(0.1 nM,10 nM)或C5a(8微克蛋白质/毫升,32微克蛋白质/毫升)进行连续刺激时,平均±1标准差的值分别为51±9%(N),30±9%(C)和34±10(N),48±14%(C)。如乳胶珠结合技术所示,在相同实验条件下,N PMN未能将黏附位点重新分布到细胞尾部;在研究的21个N样本中,33±8%(fMLP)或37±7%(C5a)的PMN发生受限的单极结合,而C值为70%(fMLP)或71%(C5a),P<0.001。当PMN与秋水仙碱(25微克/毫升)预孵育时观察到类似结果;C PMN表现出预期的黏附分数降低(与基线值相比),而N细胞则未出现,P<0.01。此外,秋水仙碱诱导的黏附位点重新分布在C样本中观察到(72±14%单极结合),在N PMN中显著减少(31±11%单极结合,P<0.001)。这些研究表明,与健康成人或母体对照相比,N PMN的CF感知机制正常。N PMN受刺激的运动减少可能在功能上与趋化刺激对细胞黏附性的调节降低有关。