Activation of guinea pig eosinophils by human recombinant IL-5. Selective priming to platelet-activating factor-acether and interference of its antagonists.
Activation of guinea pig eosinophils by human recombinant IL-5. Selective priming to platelet-activating factor-acether and interference of its antagonists.
复制标题
人重组 IL-5 激活豚鼠嗜酸性粒细胞。
DOI：
10.4049/jimmunol.147.8.2595
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发表时间：
1991
影响因子：
4.4
通讯作者：
B. Vargaftig
中科院分区：
文献类型：
--
作者：
Eliane Coëffier;Danielle Joseph;B. Vargaftig
血小板活化因子（PAF）-醋酸酯和IL-5在哮喘中作为嗜酸性粒细胞增殖、活化和/或募集介质的潜在作用使我们研究了人（h）rIL-5（hrIL-5）和PAF-醋酸酯单独或联合对豚鼠嗜酸性粒细胞的作用。在不连续的甲泛葡胺梯度下，从多粘菌素B处理的豚鼠的腹腔灌洗液中分离出两个嗜酸性粒细胞群：一个为低密度（20 - 22%的甲泛葡胺，纯度：63 +/-3%，n = 27），另一个为正常密度（22 - 24%的甲泛葡胺，纯度：87 +/-2%，n = 16）。在微型Boyden室上评价趋化活性，结果表示为在激动剂或仅溶剂存在下，通过硝酸纤维素过滤器（3微米孔径）的40微米处的迁移嗜酸性粒细胞的数量（平均值+/- SEM）。hrIL-5剂量依赖性地刺激正常密度嗜酸性粒细胞趋化性，在500 ng/ml时达到峰值（98 +/-21个迁移嗜酸性粒细胞，n = 5，p小于0.05）。这些嗜酸性粒细胞也对PAF-醋酸酯和LTB 4有反应，但对FMLP、hrTNF α和LPS无反应。嗜酸性粒细胞与hrIL-5预孵育显著增加了PAF-乙酸酯的迁移（与hrIL-5 500 ng/ml预孵育后，10 nM PAF-乙酸酯的嗜酸性粒细胞迁移率为173 +/- 23，与单独缓冲液预孵育后为69 +/- 10，p <0.01），但未能增强LTB 4的迁移或发现FMLP的活性。当细胞与拮抗剂BN 52021和WEB 2086预孵育时，PAF-乙酸的迁移被拮抗，拮抗剂BN 52021和WEB 2086也抑制hrIL-5的迁移。嗜酸性粒细胞自身脱敏和PAF-乙酰或LTB 4，但没有交叉脱敏对方。嗜酸性粒细胞对PAF-醋酸酯脱敏后不能随hrIL-5迁移，而对LTB 4脱敏的嗜酸性粒细胞作为对照对hrIL-5有反应。hrIL-5未能诱导细胞内游离钙浓度和超氧阴离子生成的基础值的升高，而与hrIL-5的嗜酸性粒细胞的预孵育诱导细胞内游离钙浓度和超氧阴离子生成的10 nM PAF-乙酸酯，但不是LTB 4的上升显着增加。总之，过敏反应中的体内嗜酸性粒细胞迁移可能涉及hrIL-5，特别是与PAF-乙酰基有关。
The potential role of platelet-activating factor (PAF)-acether and of IL-5 as an eosinophil-proliferating, activating, and/or recruiting mediator in asthma led us to study the effects of human (h) rIL-5 (hrIL-5) and PAF-acether, alone or combined, on isolated guinea pig eosinophils. Two populations of eosinophils were separated from peritoneal lavages of polymyxin B-treated guinea pigs upon a discontinuous metrizamide gradient: one of low density (between 20 and 22% of metrizamide, purity: 63 +/- 3%, n = 27) and another of normal density (between 22 and 24% of metrizamide, purity: 87 +/- 2%, n = 16). Chemotactic activity was evaluated on a micro-Boyden chamber, results being expressed as the number of migrating eosinophils (mean +/- SEM) at 40 microns through a cellulose nitrate filter (3 microns pore size) in the presence of the agonist or of the solvent alone. hrIL-5 dose-dependently stimulated normodense eosinophil chemotaxis, reaching a peak at 500 ng/ml (98 +/- 21 migrating eosinophils, n = 5, p less than 0.05). These eosinophils also responded to PAF-acether and to LTB4 and not to FMLP, hrTNF alpha, and LPS. Eosinophil preincubation with hrIL-5 increased significantly the migration by PAF-acether (173 +/- 23 migrating eosinophils with PAF-acether 10 nM after preincubation with hrIL-5 500 ng/ml vs 69 +/- 10 after preincubation with buffer alone, p less than 0.01) and failed to enhance migration by LTB4 or to uncover an activity for FMLP. Migration by PAF-acether was antagonized when the cells were preincubated with the antagonists BN 52021 and WEB 2086, which also inhibited migration by hrIL-5. Eosinophils were auto-desensitized by and to PAF-acether or LTB4, but were not cross-desensitized to each other. Eosinophils desensitized to PAF-acether failed to migrate with hrIL-5, but those desensitized to LTB4 responded to hrIL-5 as controls. hrIL-5 failed to induce the elevation of intracellular free calcium concentration and superoxide anion generation from basal values, whereas preincubation of eosinophils with hrIL-5 induced a significant increase in the rise in intracellular free calcium concentration and in superoxide anion generation by 10 nM PAF-acether but not by LTB4. In conclusion, the in vivo eosinophil migration in allergy may involve hrIL-5, particularly associated to PAF-acether.