Ryan L K, Vermeulen M W
Pulmonary and Critical Care Unit, Massachusetts General Hospital, Boston, USA.
Am J Respir Cell Mol Biol. 1995 May;12(5):540-6. doi: 10.1165/ajrcmb.12.5.7742017.
Alveolar macrophages (AM) orchestrate the release of several cytokines in the lung, including tumor necrosis factor alpha (TNF). Lipopolysaccharide endotoxin (LPS), one of the most potent stimulators of TNF production in macrophages, often contributes to the development of adult respiratory distress syndrome. The mechanism by which LPS induces TNF production in macrophages is unclear. Many studies have employed murine macrophages lavaged from the peritoneal cavity (PM), either resident cells or those obtained following elicitation with sterile thioglycollate (TGPM), as these cells are readily accessible. LPS does not induce TNF in PM or TGPM from C3H/HeJ mice, and the alteration is thought to reside at the post-transcriptional level of gene regulation. Generalization of results from PM to AM may not be warranted, however. While investigating cytokine production by AM cell lines, we observed that C3HAMSV40, a transformed cell line derived from C3H/HeJ AM, made substantial quantities of TNF when stimulated with 1 microgram/ml LPS, prompting us to study TNF production in primary C3H/HeJ AM. In the present study, readily detectable quantities of biologically active TNF were found in supernatants of C3H/HeJ AM that had been stimulated in vitro with 0.01 to 10 micrograms/ml LPS. The amount of TNF produced was not significantly different from that observed with AM from endotoxin-sensitive C3HeB/FeJ mice. LPS induction of TNF in AM from either mouse strain was completely inhibited by polymyxin B, demonstrating that the sensitivity of C3H/HeJ AM was not due to a contaminant in the LPS.(ABSTRACT TRUNCATED AT 250 WORDS)
肺泡巨噬细胞(AM)在肺中协调多种细胞因子的释放,包括肿瘤坏死因子α(TNF)。脂多糖内毒素(LPS)是巨噬细胞中TNF产生的最有效刺激物之一,常导致成人呼吸窘迫综合征的发生。LPS诱导巨噬细胞产生TNF的机制尚不清楚。许多研究采用从腹腔灌洗获得的小鼠巨噬细胞(PM),包括驻留细胞或经无菌巯基乙酸盐(TGPM)诱导后获得的细胞,因为这些细胞易于获取。LPS不会在C3H/HeJ小鼠的PM或TGPM中诱导TNF产生,这种改变被认为存在于基因调控的转录后水平。然而,将PM的结果推广到AM可能并不合理。在研究AM细胞系的细胞因子产生时,我们观察到C3HAMSV40,一种源自C3H/HeJ AM的转化细胞系,在用1微克/毫升LPS刺激时会产生大量TNF,这促使我们研究原代C3H/HeJ AM中的TNF产生。在本研究中,在体外经0.01至10微克/毫升LPS刺激的C3H/HeJ AM的上清液中发现了易于检测到的生物活性TNF量。产生的TNF量与对内毒素敏感的C3HeB/FeJ小鼠的AM中观察到的量没有显著差异。多粘菌素B完全抑制了两种小鼠品系的AM中LPS诱导的TNF产生,表明C3H/HeJ AM的敏感性不是由于LPS中的污染物所致。(摘要截断于250字)