Del Pozo V, De Andres B, Martin E, Maruri N, Zubeldia J M, Palomino P, Lahoz C
Department of Immunology, Fundación Jimenez Diaz, Hospital Gregorio Marañon, Madrid, Spain.
J Immunol. 1990 Apr 15;144(8):3117-22.
The presence of IL-1 mRNA in eosinophils from mice infected with larvae of the parasite Mesocestoides corti was investigated by in situ hybridization technique. S35 labeled cDNA probe for alpha IL-1, hybridized with mRNA in murine eosinophils and macrophages. After 6 h of LPS stimulation eosinophils were able to express mRNA in their cytoplasm. This expression was highly increased by the addition of indomethacine. The IL-1 mRNA expression in murine macrophages was higher than in eosinophils in LPS-stimulated cells. This difference was statistically significant, p less than 0.001. To test if eosinophils may produce and release IL-1 in the culture medium, we isolated these cells in a Percoll gradient. Cell preparations with a purity exceeding 94% were cultured with various stimuli and their supernatants were tested for IL-1 activity. Eosinophils produced 169.65 +/- 73 U/ml when stimulated with LPS (n = 14). A dose-dependent response was obtained when the eosinophils were in the presence of the calcium ionophore A23187. Controls were performed to rule out the contribution of the contaminating population on the thymocyte proliferating activity. They were also used to detect other possible causes of interference in the assay, such as leukotrienes or TNF. IL-1 in supernatants was also detected using a conversion assay such as EL-4 thymoma cells. IL-1 activity was first detected in culture supernatants 18 h later, maximal production being in the first 24 h. In accordance with our hybridization results, an increase in IL-1 activity was obtained when eosinophils were stimulated with LPS and treated with indomethacine. The factor had a molecular mass between 16 to 20 kDa that corresponded to the described for murine IL-1. Inasmuch as IL-1 is an important mediator of inflammatory reactions this IL may enhance the proinflammatory action of eosinophils.
采用原位杂交技术研究了感染寄生虫中殖孔绦虫幼虫的小鼠嗜酸性粒细胞中白细胞介素-1（IL-1）信使核糖核酸（mRNA）的存在情况。用α IL-1的S35标记互补DNA（cDNA）探针与小鼠嗜酸性粒细胞和巨噬细胞中的mRNA杂交。脂多糖（LPS）刺激6小时后，嗜酸性粒细胞能够在其细胞质中表达mRNA。吲哚美辛的加入使这种表达显著增加。在LPS刺激的细胞中，小鼠巨噬细胞中IL-1 mRNA的表达高于嗜酸性粒细胞。这种差异具有统计学意义，P小于0.001。为了检测嗜酸性粒细胞是否可能在培养基中产生并释放IL-1，我们在Percoll梯度中分离这些细胞。将纯度超过94%的细胞制剂用各种刺激物培养，并检测其上清液中的IL-1活性。用LPS刺激时，嗜酸性粒细胞产生169.65±73 U/ml（n = 14）。当嗜酸性粒细胞存在钙离子载体A23187时，获得了剂量依赖性反应。进行对照以排除污染群体对胸腺细胞增殖活性的影响。它们还用于检测测定中其他可能的干扰原因，如白三烯或肿瘤坏死因子（TNF）。还用诸如EL-4胸腺瘤细胞的转化测定法检测上清液中的IL-1。18小时后在培养上清液中首次检测到IL-1活性，最大产量在最初24小时。根据我们的杂交结果，当嗜酸性粒细胞用LPS刺激并用吲哚美辛处理时，IL-1活性增加。该因子的分子量在16至20 kDa之间，与所述的小鼠IL-1相对应。由于IL-1是炎症反应的重要介质，这种IL可能增强嗜酸性粒细胞的促炎作用。
