Sugar A M, Field K G
Evans Memorial Department of Clinical Research, University Hospital, Boston 02118.
J Leukoc Biol. 1988 Dec;44(6):500-7. doi: 10.1002/jlb.44.6.500.
Stimulation of the respiratory burst of murine bronchoalveolar macrophages obtained by lung lavage was studied using four different stimuli and different assay conditions. One soluble stimulus, phorbol myristate acetate (PMA), two intracellular particles, zymosan and Blastomyces dermatitidis conida, and one extracellular particle, B. dermatitidis yeast, were incubated with either freshly obtained macrophages in suspension or 2- and 48-hour macrophage monolayers. Suspension cultures were incubated with stimuli for 90 minutes and monolayers for 10 minutes before O2- was assayed. PMA did not elicit O2- production in macrophage suspensions or 2-hour macrophage monolayers, but 48-hour macrophage monolayers exhibited a 13-fold increase above control values (P = .0001). On the other hand, zymosan elicited an increase in O2- production in both suspensions and monolayers, although monolayers incubated for 48 hours produced almost fourfold more O2- than the other systems (P = .025). Opsonization had no effect on the ability of zymosan to elicit respiratory burst. B. dermatitidis conidia resulted in a two- to threefold increase in O2- production in macrophage suspensions and a five- to eightfold increase in 48-hour monolayers, representing significantly less respiratory burst stimulation than with either zymosan or PMA. Similarly, B. dermatitidis yeasts demonstrated similar submaximal stimulation of O2-, 3-4 times that over control values, and again this was less than zymosan and PMA. We conclude that 1) freshly obtained murine bronchoalveolar macrophages do not respond to PMA with an increase in O2- production, but that responsiveness is evident after 48 hours of incubation in monolayers; 2) B. dermatitidis conidia and yeasts do not stimulate respiratory burst activity to the same degree as zymosan or PMA; and 3) opsonization of zymosan is not necessary for stimulation of the murine bronchoalveolar macrophage oxidative burst, confirming previous data that functional complement receptors are not present on these cells.
使用四种不同的刺激物和不同的检测条件，研究了通过肺灌洗获得的小鼠支气管肺泡巨噬细胞呼吸爆发的刺激情况。一种可溶性刺激物，佛波酯肉豆蔻酸酯乙酸盐（PMA），两种细胞内颗粒，酵母聚糖和皮炎芽生菌分生孢子，以及一种细胞外颗粒，皮炎芽生菌酵母，分别与新鲜获得的悬浮巨噬细胞或培养2小时和48小时的巨噬细胞单层进行孵育。在检测超氧阴离子（O2-）之前，悬浮培养物与刺激物孵育90分钟，单层培养物孵育10分钟。PMA在巨噬细胞悬浮液或培养2小时的巨噬细胞单层中未引起O2-产生，但培养48小时的巨噬细胞单层显示出比对照值增加13倍（P = 0.0001）。另一方面，酵母聚糖在悬浮液和单层中均引起O2-产生增加，尽管培养48小时的单层产生的O2-几乎是其他系统的四倍（P = 0.025）。调理作用对酵母聚糖引发呼吸爆发的能力没有影响。皮炎芽生菌分生孢子导致巨噬细胞悬浮液中O2-产生增加两到三倍，在培养48小时的单层中增加五到八倍，这表明与酵母聚糖或PMA相比，其对呼吸爆发的刺激明显较小。同样，皮炎芽生菌酵母对O2-的刺激也类似，是对照值的3 - 4倍，同样低于酵母聚糖和PMA。我们得出结论：1）新鲜获得的小鼠支气管肺泡巨噬细胞对PMA刺激不会引起O2-产生增加，但在单层培养48小时后反应明显；2）皮炎芽生菌分生孢子和酵母对呼吸爆发活性的刺激程度不如酵母聚糖或PMA；3）酵母聚糖的调理作用对于刺激小鼠支气管肺泡巨噬细胞氧化爆发不是必需的，这证实了先前的数据，即这些细胞上不存在功能性补体受体。
