Ying S, Meng Q, Taborda-Barata L, Corrigan C J, Barkans J, Assoufi B, Moqbel R, Durham S R, Kay A B
National Heart and Lung Institute, London, England, GB.
Eur J Immunol. 1996 Jan;26(1):70-6. doi: 10.1002/eji.1830260111.
Eosinophils synthesize and store various cytokines with potential autocrine activity. We hypothesized that eosinophils synthesize and store RANTES, a CC-chemokine with potent eosinophil chemotactic activity. Expression of RANTES mRNA in highly purified eosinophil populations was detected by reverse transcription followed by polymerase chain reaction analysis. In situ hybridization (ISH) with 35S-labeled RANTES-specific riboprobes showed that 6.8-10% of peripheral blood eosinophils obtained from atopic subjects expressed RANTES mRNA, increasing to 25% after incubation (16 h) with interferon (IFN)-gamma, but not ionomycin in vitro. Peripheral blood eosinophils also showed specific immunoreactivity with an anti-RANTES monoclonal antibody, consistent with translation of the mRNA. By enzyme-linked immunosorbent assay, blood eosinophils were shown to contain a median of 7300 pg (range 5200-8800) RANTES per 10(6) cells, of which a mean of 24% was released into culture supernatants after stimulation of the cells with serum-coated particles in vitro. These culture supernatants exhibited eosinophil chemotactic activity which was inhibited (mean 68%) by a specific anti-RANTES antibody. Sequential immunocytochemistry and ISH on biopsies obtained from allergen-induced late-phase cutaneous reactions showed that 55-75% of the infiltrating RANTES mRNA+ cells were EG2+ eosinophils. Allergen, but not diluent challenge, was also associated with a time-dependent increase in the number of cells showing RANTES immunoreactivity. Of these cells, 55% were identified as eosinophils by morphological criteria. Thus, human eosinophils have the capacity to synthesize, store and secrete physiologically relevant quantities of RANTES, and may therefore be an important source of this chemokine in allergic inflammation.
嗜酸性粒细胞合成并储存具有潜在自分泌活性的多种细胞因子。我们推测嗜酸性粒细胞合成并储存RANTES，一种具有强大嗜酸性粒细胞趋化活性的CC趋化因子。通过逆转录随后进行聚合酶链反应分析来检测高度纯化的嗜酸性粒细胞群体中RANTES mRNA的表达。用35S标记的RANTES特异性核糖探针进行原位杂交（ISH）显示，从特应性受试者获得的外周血嗜酸性粒细胞中有6.8 - 10%表达RANTES mRNA，在体外与干扰素（IFN）-γ孵育（16小时）后增加到25%，但与离子霉素孵育后无增加。外周血嗜酸性粒细胞也显示出与抗RANTES单克隆抗体的特异性免疫反应性，这与mRNA的翻译一致。通过酶联免疫吸附测定，显示每10(6)个血液嗜酸性粒细胞中RANTES的中位数含量为7300 pg（范围5200 - 8800），其中在用血清包被颗粒体外刺激细胞后，平均有24%释放到培养上清液中。这些培养上清液表现出嗜酸性粒细胞趋化活性，该活性被特异性抗RANTES抗体抑制（平均68%）。对变应原诱导的迟发性皮肤反应活检组织进行连续免疫细胞化学和ISH显示，浸润的RANTES mRNA+细胞中有55 - 75%是EG2+嗜酸性粒细胞。变应原而非稀释剂激发也与显示RANTES免疫反应性的细胞数量随时间增加有关。在这些细胞中，根据形态学标准55%被鉴定为嗜酸性粒细胞。因此，人类嗜酸性粒细胞有能力合成、储存和分泌生理相关量的RANTES，因此可能是变应性炎症中这种趋化因子的重要来源。
