Eosinophils altered phenotypically and primed by culture with granulocyte/macrophage colony-stimulating factor and 3T3 fibroblasts generate leukotriene C4 in response to FMLP.
Eosinophils altered phenotypically and primed by culture with granulocyte/macrophage colony-stimulating factor and 3T3 fibroblasts generate leukotriene C4 in response to FMLP.
复制标题
嗜酸性粒细胞表型发生改变，并通过粒细胞/巨噬细胞集落刺激因子培养引发，3T3 成纤维细胞响应 FMLP 产生白三烯 C4。
DOI：
10.1172/jci115222
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Austen,KF
中科院分区：
文献类型：
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作者：
OwenJr,WF;Petersen,J;Austen,KF
正常密度的嗜酸性粒细胞未能产生白三烯C4（LTC 4）在响应FMLP的递增浓度，但产生LTC 4时，刺激钙离子载体A23187。正常密度的嗜酸性粒细胞，在3 T3成纤维细胞存在下，与10（-11）M粒细胞/巨噬细胞集落刺激因子（GM-CSF）一起培养，在第4天对FMLP的跨膜刺激产生反应，在第7天达到最大效应。培养7天后，用2 × 10（-7）M FMLP刺激低密度嗜酸性粒细胞产生26 ng LTC 4/10（6）细胞，LTC 4生物合成被N-叔丁氧基羰基-L-甲硫氨酰-L-亮氨酰-L-苯丙氨酸（N-t-BOC-MLP）阻断。当嗜酸性粒细胞与GM-CSF和3 T3成纤维细胞共培养时，无论是内源性花生四烯酸对LTC 4的钙离子载体刺激，还是掺入LTA 4的底物启动的LTC 4产生都没有改变。此外，当与10（-6）M FMLP孵育时，正常密度的嗜酸性粒细胞不产生净超氧化物，通过细胞色素c的还原来测量，而与10（-11）M GM-CSF和3 T3成纤维细胞孵育7天的重复嗜酸性粒细胞减少了17 nmol细胞色素c/10（6）细胞的净超氧化物。这些研究表明，致敏和表型改变的嗜酸性粒细胞存在于血管外的网站可能会发挥病理生物学效应，通过响应组织中的可溶性配体。
Normodense eosinophils failed to generate leukotriene C4 (LTC4) in response to incremental concentrations of FMLP but did produce LTC4 when stimulated with calcium ionophore A23187. Normodense eosinophils, maintained in culture with 10(-11) M granulocyte/macrophage colony-stimulating factor (GM-CSF) in the presence of 3T3 fibroblasts, became responsive to transmembrane stimulation with FMLP by day 4 with a maximal effect by day 7. After 7 d of culture, hypodense eosinophils stimulated with 2 x 10(-7) M FMLP generated 26 ng LTC4/10(6) cells, and LTC4 biosynthesis was blocked by N-tertbutoxy-carbonyl-L-methionyl-L-leucyl-L-phenylalanine (N-t-BOC-MLP). Neither calcium ionophore stimulation of LTC4 from endogenous arachidonic acid nor substrate-initiated production of LTC4 from incorporated LTA4 changed when eosinophils were cocultured with GM-CSF and 3T3 fibroblasts. Furthermore, when incubated with 10(-6) M FMLP, normodense eosinophils generated no net superoxide measured by the reduction of cytochrome c, whereas replicate eosinophils cultured for 7 d with 10(-11) M GM-CSF and 3T3 fibroblasts reduced a net of 17 nmol of cytochrome c/10(6) cells. These studies suggest that primed and phenotypically altered eosinophils present at an extravascular site may exert pathobiologic effects by responding to soluble ligands in the tissues.