Goodrum K J
J Leukoc Biol. 1987 Apr;41(4):295-301. doi: 10.1002/jlb.41.4.295.
Secretion of complement component C3 by the mouse macrophage-like cell lines PU5-1.8, J774A.1, RAW264.7, and P388D1 was measured using an enzyme-linked immunosorbent assay for mouse C3. All cell lines secreted antigenically detectable C3 with the relative secreted C3/10(6) cells/24 h ranked as J774A.1 greater than P388D1 greater than or equal to PU5-1.8 much greater than RAW264.7. C3 secretion was enhanced two- to fourfold in cultures of all cell lines when treated with lipopolysaccharide, streptococcal cell walls, or lymphokine-containing supernatant fluids of mitogen-stimulated spleen cells. A differential induction of C3 synthesis and secretion was indicated since secreted lysozyme and total cellular protein were not elevated in a manner comparable to C3. The relative inducibility of cell lines for C3 secretion in either lipopolysaccharide- or cell wall-treated cells could be ranked as PU5-1.8 greater than P388D1 greater than J774A.1 greater than RAW264.7. C3 secretion was inhibited by cycloheximide or hydrocortisone. Mouse macrophage-like cell lines retain baseline and inducible C3 synthetic activities as do normal macrophages and can serve as homogeneous cultures in which to study regulation of complement biosynthesis.
使用针对小鼠C3的酶联免疫吸附测定法,检测了小鼠巨噬细胞样细胞系PU5-1.8、J774A.1、RAW264.7和P388D1中补体成分C3的分泌情况。所有细胞系均分泌可通过抗原检测到的C3,每10⁶个细胞/24小时相对分泌的C3量排序为:J774A.1 > P388D1 ≥ PU5-1.8 >> RAW264.7。当用脂多糖、链球菌细胞壁或丝裂原刺激的脾细胞的含淋巴因子的上清液处理时,所有细胞系培养物中的C3分泌增强了2至4倍。由于分泌的溶菌酶和总细胞蛋白没有以与C3相当的方式升高,表明存在C3合成和分泌的差异诱导。在脂多糖或细胞壁处理的细胞中,细胞系C3分泌的相对诱导能力排序为:PU5-1.8 > P388D1 > J774A.1 > RAW264.7。环己酰亚胺或氢化可的松可抑制C3分泌。小鼠巨噬细胞样细胞系与正常巨噬细胞一样,保留了基线和可诱导的C3合成活性,并且可以作为研究补体生物合成调节的同质培养物。