Nordlund J J, Root R K, Wolff S M
J Exp Med. 1970 Apr 1;131(4):727-43. doi: 10.1084/jem.131.4.727.
Release of the protein molecule, leukocytic pyrogen, is one of the many reactions exhibited by leukocytes after phagocytosis. After the ingestion of heat-killed S. albus, a 3-4 hr latent period exists, during which human peripheral leukocytes release no pyrogen, yet cellular metabolism is altered in such a way that pyrogen output may subsequently occur in the absence of further phagocytosis. Transcription of messenger RNA and translation of new protein are initial events in the. activation process, since addition of the inhibitors, actinomycin D, and cycloheximide or puromycin, during this period markedly depressed or abolished subsequent pyrogen release. These effects were noted to be dependent upon the time of addition of the inhibitors. None of the inhibitor drugs interfered with cell viability as measured by phagocytosis and hexose monophosphate shunt activity, nor did they alter the pyrogenicity of preformed leukocytic pyrogen. Vincristine did not inhibit pyrogen formation, consistent with its reported failure to alter RNA synthesis in mature human granulocytes. The glycolytic inhibitor, sodium fluoride, blocked pyrogen release both when added prior to particle ingestion or 1 hr after the initiation of phagocytosis. Whereas inhibition of phagocytosis would explain the sodium fluoride effect prior to 1 hr, this was not observed in leukocyte preparations incubated for 1 hr with S. albus before adding sodium fluoride. When sodium fluoride was added to preparations 2 hr after the start of incubation, the LP production was unimpaired. Potassium cyanide had no effect on cell activation or pyrogen release. These findings suggest that the primary energy supply for the activation process is derived from high energy phosphate bonds provided by anaerobic glycolysis. Since the major amount of cell activation appears to occur in the 1st hr after phagocytosis, this energy might be involved in the induction of a genome leading to the transcription of m-RNA and its translation into new protein or is required for polysome integrity during protein synthesis. It is suggested that this new protein may be leukocytic pyrogen itself, or an enzyme responsible for cleaving it from an inactive precursor.
蛋白质分子——白细胞致热原的释放，是白细胞吞噬作用后出现的众多反应之一。摄入热杀死的白色链霉菌后，有3至4小时的潜伏期，在此期间，人外周血白细胞不释放致热原，但细胞代谢发生改变，以至于在没有进一步吞噬作用的情况下随后可能产生致热原输出。信使核糖核酸的转录和新蛋白质的翻译是激活过程中的初始事件，因为在此期间添加抑制剂放线菌素D、环己酰亚胺或嘌呤霉素会显著抑制或消除随后的致热原释放。注意到这些作用取决于抑制剂添加的时间。这些抑制药物均未干扰通过吞噬作用和磷酸己糖旁路活性测定的细胞活力，它们也未改变预先形成的白细胞致热原的致热能力。长春新碱不抑制致热原的形成，这与其报道的未能改变成熟人粒细胞中RNA合成一致。糖酵解抑制剂氟化钠，在颗粒摄入前或吞噬作用开始后1小时添加时，均能阻断致热原的释放。虽然吞噬作用的抑制可以解释氟化钠在1小时前的作用，但在加入氟化钠之前用白色链霉菌孵育1小时的白细胞制剂中未观察到这种情况。当在孵育开始后2小时向制剂中添加氟化钠时，白细胞致热原的产生未受损害。氰化钾对细胞激活或致热原释放没有影响。这些发现表明，激活过程的主要能量供应来自无氧糖酵解提供的高能磷酸键。由于大量的细胞激活似乎发生在吞噬作用后的第1小时，这种能量可能参与诱导基因组导致信使核糖核酸的转录及其翻译成新蛋白质，或者是蛋白质合成过程中多核糖体完整性所必需的。有人认为，这种新蛋白质可能本身就是白细胞致热原，或者是负责从无活性前体中切割它的一种酶。
