Shaw R J, Cromwell O, Kay A B
Clin Exp Immunol. 1984 Jun;56(3):716-22.
The leukotriene generating capacities of ionophore stimulated human eosinophils and neutrophils were compared using specific radioimmunoassays for LTB4 and LTC4. Mixed granulocyte preparations (neutrophils and eosinophils) produced both LTB4 and LTC4 in a time-dependent fashion which was maximal at 10 and 15 min, respectively. Following the separation of eosinophils (greater than 75%) and neutrophils (greater than 90%) by metrizamide gradients, LTC4 production was predominantly from eosinophils, whereas neutrophils were the principal source of LTB4. The concentrations of leukotrienes produced by the eosinophil and neutrophil rich cell preparations were directly proportional to the concentration of ionophore. Following purification of eosinophil derived products by RP-HPLC the LTC4 immunoreactivity corresponded to the elution profile of a synthetic LTC4 marker. Furthermore, in 32 atopic subjects (21 bronchial asthmatics and 11 non-asthmatics) the amounts of LTC4 produced by unseparated leucocytes were directly proportional to the percentage of eosinophils in the total cell suspension. Preferential generation of LTB4 by neutrophils was also demonstrated by immunoreactivity of ionophore stimulated supernatants subjected to RP-HPLC, as well as by its characteristic u.v. absorbance and GC-MS profile and the ability to promote directional neutrophil locomotion (chemotaxis). These experiments support the concept that eosinophils accumulate in tissues partly as a result of the response to neutrophil derived LTB4, and that these cells contribute to the production of sulphidopeptide leukotrienes with subsequent amplification of the acute allergic response.
利用针对白三烯B4(LTB4)和白三烯C4(LTC4)的特异性放射免疫分析法,比较了离子载体刺激的人嗜酸性粒细胞和中性粒细胞生成白三烯的能力。混合粒细胞制剂(中性粒细胞和嗜酸性粒细胞)以时间依赖性方式产生LTB4和LTC4,分别在10分钟和15分钟时达到最大值。通过甲泛葡胺梯度分离嗜酸性粒细胞(大于75%)和中性粒细胞(大于90%)后,LTC4主要由嗜酸性粒细胞产生,而中性粒细胞是LTB4的主要来源。富含嗜酸性粒细胞和中性粒细胞的细胞制剂产生的白三烯浓度与离子载体的浓度成正比。通过反相高效液相色谱法(RP-HPLC)纯化嗜酸性粒细胞衍生产物后,LTC4免疫反应性与合成LTC4标志物的洗脱曲线一致。此外,在32名特应性受试者(21名支气管哮喘患者和11名非哮喘患者)中,未分离的白细胞产生的LTC4量与总细胞悬液中嗜酸性粒细胞的百分比成正比。通过对经RP-HPLC处理的离子载体刺激上清液的免疫反应性,以及其特征性紫外吸收、气相色谱-质谱(GC-MS)图谱和促进中性粒细胞定向运动(趋化性)的能力,也证明了中性粒细胞优先产生LTB4。这些实验支持了这样的概念,即嗜酸性粒细胞在组织中积聚部分是对中性粒细胞衍生的LTB4反应的结果,并且这些细胞有助于硫肽白三烯的产生,随后放大急性过敏反应。