Czop J K, Puglisi A V, Miorandi D Z, Austen K F
Department of Medicine, Harvard Medical School, Boston, MA 02115.
J Immunol. 1988 Nov 1;141(9):3170-6.
Normal human neutrophils were stimulated with the yeast cell wall product, zymosan, and examined for two biologic responses, ingestion of particles and production of leukotriene B4 (LTB4), under conditions that were comparable and optimal for the quantitation of each response. Monolayers of adherent neutrophils ingested unopsonized zymosan particles, at particle-to-cell ratios of 12.5:1 to 125:1, in a dose- and time-related manner. At a ratio of 125:1, the percentages of neutrophils ingesting greater than or equal to 1 and greater than or equal to 3 zymosan particles reached plateau levels of 55 +/- 6 and 32 +/- 9% (mean +/- SD, n = 8), respectively, within 30 min. At this same ratio, neutrophils during gravity sedimentation with zymosan particles synthesized LTB4 in a time-dependent manner for at least 45 min. The maximum amount of immunoreactive LTB4 released into supernatants was 3.8 +/- 1.2 ng per 10(6) neutrophils (mean +/- SD, n = 5) and the corresponding total immunoreactive LTB4 was 6.2 +/- 1.9 ng per 10(6) neutrophils. Treatment of 2 x 10(7) suspended neutrophils with 250 micrograms of trypsin for 20 min before concurrent assessment of neutrophil phagocytosis and LTB4 production reduced both of these responses by about 50%. Pretreatment of neutrophils with 800 micrograms/ml of soluble yeast beta-glucan inhibited their ingestion of zymosan by 84% (mean +/- SD, n = 3), with 50% inhibition occurring with 100 micrograms/ml of soluble beta-glucan; 800 micrograms/ml of soluble yeast alpha-mannan had no inhibitory effect. Pretreatment of neutrophils with 400 micrograms/ml of soluble yeast beta-glucan inhibited neutrophil synthesis of LTB4 by 90%, with 50% occurring with 200 micrograms/ml; 400 micrograms/ml of soluble yeast alpha-mannan had no inhibitory effect. The presence of 1.25 micrograms/ml of cytochalasin B during incubation with zymosan particles reduced neutrophil phagocytosis from 65 to 6%, and neutrophil synthesis of LTB4 from total levels of 6.0 +/- 0.3 ng/10(6) cells to zero (mean +/- SD, n = 3). Pretreatment with either cytochalasin B or vinblastine did not alter neutrophil generation of LTB4 induced by calcium ionophore. Neutrophils pretreated with vinblastine, at 4 x 10(-6) to 4 x 10(-4) M, and then maintained at one-half these concentrations during incubation with unopsonized zymosan particles exhibited no diminution in particle ingestion, but were markedly reduced in zymosan-induced synthesis of LTB4.(ABSTRACT TRUNCATED AT 400 WORDS)
用酵母细胞壁产物酵母聚糖刺激正常人中性粒细胞，并在对每种反应进行定量分析的可比且最佳条件下，检测两种生物学反应，即颗粒摄取和白三烯B4（LTB4）的产生。贴壁中性粒细胞单层以12.5:1至125:1的颗粒与细胞比例摄取未调理的酵母聚糖颗粒，呈剂量和时间依赖性。在125:1的比例下，摄取大于或等于1个以及大于或等于3个酵母聚糖颗粒的中性粒细胞百分比在30分钟内分别达到55±6%和32±9%的平台水平（平均值±标准差，n = 8）。在相同比例下，与酵母聚糖颗粒进行重力沉降的中性粒细胞以时间依赖性方式合成LTB4至少45分钟。释放到上清液中的免疫反应性LTB4的最大量为每10^6个中性粒细胞3.8±1.2 ng（平均值±标准差，n = 5），相应的总免疫反应性LTB4为每10^6个中性粒细胞6.2±1.9 ng。在同时评估中性粒细胞吞噬作用和LTB4产生之前，用250微克胰蛋白酶处理2×10^7个悬浮中性粒细胞20分钟，这两种反应均降低了约50%。用800微克/毫升的可溶性酵母β-葡聚糖预处理中性粒细胞可使其对酵母聚糖的摄取抑制84%（平均值±标准差，n = 3），100微克/毫升的可溶性β-葡聚糖可产生50%的抑制作用；800微克/毫升可溶性酵母α-甘露聚糖无抑制作用。用400微克/毫升的可溶性酵母β-葡聚糖预处理中性粒细胞可使其LTB4合成抑制90%，200微克/毫升时产生50%的抑制作用；400微克/毫升可溶性酵母α-甘露聚糖无抑制作用。在与酵母聚糖颗粒孵育期间存在1.25微克/毫升的细胞松弛素B可使中性粒细胞吞噬作用从65%降至6%，中性粒细胞LTB4合成从总水平6.0±0.3 ng/10^6个细胞降至零（平均值±标准差，n = 3）。用细胞松弛素B或长春碱预处理均未改变钙离子载体诱导的中性粒细胞LTB4生成。用4×10^-6至4×10^-4 M的长春碱预处理中性粒细胞，然后在与未调理的酵母聚糖颗粒孵育期间维持在这些浓度的一半，颗粒摄取无减少，但酵母聚糖诱导的LTB4合成明显减少。（摘要截短至400字）
