Hirata K, Pelé J P, Robidoux C, Sirois P
Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
J Leukoc Biol. 1989 Jun;45(6):523-8. doi: 10.1002/jlb.45.6.523.
Guinea pig lung eosinophils have been purified from bronchoalveolar lavage (BAL) fluid, and the production of prostaglandin E2 (PGE2), thromboxane B2 (TXB2), and 6-keto-PGF1 alpha was measured after stimulation with phorbol myristate acetate (PMA), platelet-activating factor (PAF), and formyl-methionyl-leucyl-phenylalanine (fMLP). A combination of plating and discontinuous Percoll gradient centrifugation was used to purify eosinophils. The purity of eosinophils in fraction E (interface between Percoll density 1.057-1.068) was around 96%, and the viability was 99%, with a mean yield of 2.7 x 10(6) cells per guinea pig. Similarly, in fraction D (interface between Percoll density 1.047-1.057), the mean purity of eosinophils was 76% and the viability of cells was 99%, with a mean yield of 1.1 x 10(6) cells per guinea pig. Purified eosinophils produced TXB2 predominantly after stimulation with PMA, fMLP, and PAF. The 6-keto-PGF1 alpha was slightly increased in cell supernatants after stimulation with PMA and fMLP, but PGE2 was not elevated with any stimulus. The highly purified eosinophils of fraction E generated amounts of TXB2 and 6-keto-PGF1 alpha similar to the amount generated by eosinophils contaminated with some macrophages (fraction D). These results suggest a role for eosinophils in the production of TXA2 by guinea pig lung.
豚鼠肺嗜酸性粒细胞已从支气管肺泡灌洗(BAL)液中纯化出来,在用佛波酯肉豆蔻酸酯(PMA)、血小板活化因子(PAF)和甲酰甲硫氨酰亮氨酰苯丙氨酸(fMLP)刺激后,检测了前列腺素E2(PGE2)、血栓素B2(TXB2)和6-酮-前列腺素F1α的产生。采用铺板和不连续 Percoll 梯度离心相结合的方法纯化嗜酸性粒细胞。E 组分(Percoll 密度 1.057 - 1.068 之间的界面)中嗜酸性粒细胞的纯度约为 96%,活力为 99%,每只豚鼠的平均产量为 2.7×10⁶ 个细胞。同样,在 D 组分(Percoll 密度 1.047 - 1.057 之间的界面)中,嗜酸性粒细胞的平均纯度为 76%,细胞活力为 99%,每只豚鼠的平均产量为 1.1×10⁶ 个细胞。纯化后的嗜酸性粒细胞在受到 PMA、fMLP 和 PAF 刺激后主要产生 TXB2。在用 PMA 和 fMLP 刺激后,细胞上清液中的 6-酮-前列腺素F1α 略有增加,但在任何刺激下 PGE2 均未升高。E 组分中高度纯化的嗜酸性粒细胞产生的 TXB2 和 6-酮-前列腺素F1α 的量与被一些巨噬细胞污染的嗜酸性粒细胞(D 组分)产生的量相似。这些结果表明嗜酸性粒细胞在豚鼠肺中参与血栓素A2的产生。