Fearns C, Kravchenko V V, Ulevitch R J, Loskutoff D J
Department of Vascular Biology, Scripps Research Institute, La Jolla, California 92037.
J Exp Med. 1995 Mar 1;181(3):857-66. doi: 10.1084/jem.181.3.857.
A murine model system was used to study the distribution and regulation of CD14 gene expression in vivo. Western blot analysis failed to detect CD14 in plasma from untreated CB6 (BALB/c x C57Bl6) mice, but showed markedly increased levels of CD14 in plasma from mice treated with lipopolysaccharide (LPS). Plasma levels of CD14 increased in a time- and dose-dependent manner, reaching a maximum between 8 and 16 h. Northern blot analysis of total RNA extracted from mouse tissues revealed low, but significant, levels of CD14 mRNA in many tissues of untreated animals with the highest levels in uterus, adipose tissue, and lung. After intraperitoneal injection of LPS, induction of CD14 gene expression was detected in all organs examined with the extent of induction varying between organs. Induction of CD14 mRNA was both time and dose dependent. Maximum induction in the heart and lung was observed 2-4 h after injection of LPS, while liver and kidney showed maximal induction between 8 and 16 h. In situ hybridization showed that CD14 mRNA was expressed in myeloid cells in many tissues, and that expression in these cells was upregulated by LPS. Unexpectedly, CD14 mRNA was also detected in other cells within tissues, including epithelial cells, and expression in these cell types also was upregulated by LPS. Immunochemical analysis revealed that CD14 antigen colocalized to the cytoplasm of cells expressing CD14 mRNA. These studies demonstrate that CD14 gene expression is not restricted to myeloid cells, and that the level of expression of CD14 is influenced by exposure to LPS.
利用小鼠模型系统研究体内CD14基因表达的分布及调控。蛋白质免疫印迹分析未能在未处理的CB6(BALB/c×C57Bl6)小鼠血浆中检测到CD14,但脂多糖(LPS)处理的小鼠血浆中CD14水平显著升高。CD14的血浆水平呈时间和剂量依赖性增加,在8至16小时达到峰值。对从小鼠组织中提取的总RNA进行Northern印迹分析显示,未处理动物的许多组织中CD14 mRNA水平较低但显著,子宫、脂肪组织和肺中水平最高。腹腔注射LPS后,在所有检测的器官中均检测到CD14基因表达的诱导,不同器官的诱导程度有所不同。CD14 mRNA的诱导呈时间和剂量依赖性。注射LPS后2至4小时在心脏和肺中观察到最大诱导,而肝脏和肾脏在8至16小时显示最大诱导。原位杂交表明,CD14 mRNA在许多组织的髓样细胞中表达,这些细胞中的表达被LPS上调。出乎意料的是,在组织内的其他细胞(包括上皮细胞)中也检测到CD14 mRNA,这些细胞类型中的表达也被LPS上调。免疫化学分析显示,CD14抗原与表达CD14 mRNA的细胞的细胞质共定位。这些研究表明,CD14基因表达不限于髓样细胞,并且CD14的表达水平受LPS暴露的影响。