Otterness I G, Bliven M L, Eskra J D, Reinke M, Hanson D C
Department of Immunology and Infectious Diseases, Pfizer Central Research, Groton, Connecticut 06340.
Cell Immunol. 1988 Jul;114(2):385-97. doi: 10.1016/0008-8749(88)90330-9.
The role of prostaglandins in the regulation of lipopolysaccharide (LPS)-induced interleukin-1 (IL-1) production by murine C3H/HeN resident peritoneal macrophages was studied. IL-1 production was initially studied in the presence of piroxicam and indomethacin, both inhibitors of prostaglandin biosynthesis. IL-1 was assayed using the IL-1-dependent proliferative response of C3H/HeJ thymocytes. LPS stimulation resulted in 15 to 20 ng/ml of prostaglandin E2 (PGE2) produced in the first hour of culture. IL-1-containing supernatants from drug-treated macrophages at dilutions of up to 1:32 resulted in enhanced thymocyte proliferation compared to control, non-drug-treated cultures and contained less than 2 ng/ml of PGE2. Similar enhancement of proliferation could be obtained by incubating non-drug-treated supernatants with monoclonal anti-PGE2 but not anti-thromboxane B2 (TxB2) antibody. Further dilutions of the drug-treated supernatants gave thymocyte proliferation responses which were indistinguishable from control cultures and, correspondingly, had identical values for IL-1 production. The absence of an effect on IL-1 production was confirmed by quantitation of intracellular IL-1 alpha using goat anti-IL-1 alpha antibody and by quantitation of supernatant IL-1 receptor competition assay. Exogenous PGE2, in the concentration range produced in macrophage supernatants (10-20 ng/ml), directly inhibited IL-1-stimulated thymocyte proliferation. Finally, when macrophages were stimulated with LPS for 24 hr in the presence of added PGE2, thymocyte proliferation was inhibited at the lowest supernatant dilutions, but as the IL-1-containing supernatants were diluted out, the assay curves were indistinguishable from non-PGE2-treated control. Thus, in this system, PGE2 has no effect on IL-1 synthesis, but rather has a direct inhibitory effect on thymocyte proliferation. Nonsteroidal anti-inflammatory drugs are not stimulating IL-1 production but are, in fact, relieving inhibition of the thymocyte IL-1 assay caused by the presence of prostaglandins.
研究了前列腺素在调节小鼠C3H/HeN常驻腹膜巨噬细胞脂多糖（LPS）诱导的白细胞介素-1（IL-1）产生中的作用。最初在存在吡罗昔康和吲哚美辛（两者均为前列腺素生物合成抑制剂）的情况下研究IL-1的产生。使用C3H/HeJ胸腺细胞的IL-1依赖性增殖反应来测定IL-1。LPS刺激导致培养的第一小时产生15至20 ng/ml的前列腺素E2（PGE2）。与未用药物处理的对照培养物相比，来自药物处理的巨噬细胞的含IL-1的上清液在高达1:32的稀释度下导致胸腺细胞增殖增强，且含有少于2 ng/ml的PGE2。通过用单克隆抗PGE2抗体而非抗血栓素B2（TxB2）抗体孵育未用药物处理的上清液，可获得类似的增殖增强。药物处理的上清液的进一步稀释产生的胸腺细胞增殖反应与对照培养物无差异，相应地，IL-1产生的值相同。使用山羊抗IL-1α抗体对细胞内IL-1α进行定量以及通过上清液IL-1受体竞争测定法证实了对IL-1产生无影响。巨噬细胞上清液中产生的浓度范围内的外源性PGE2（10 - 20 ng/ml）直接抑制IL-1刺激的胸腺细胞增殖。最后，当在添加PGE2的情况下用LPS刺激巨噬细胞24小时时，在最低上清液稀释度下胸腺细胞增殖受到抑制，但随着含IL-1的上清液被稀释，测定曲线与未用PGE2处理的对照无差异。因此，在该系统中，PGE2对IL-1合成无影响，而是对胸腺细胞增殖具有直接抑制作用。非甾体抗炎药并非刺激IL-1产生，实际上是缓解了由前列腺素的存在对胸腺细胞IL-1测定的抑制。
