Rouzer C A, Scott W A, Hamill A L, Liu F T, Katz D H, Cohn Z A
Proc Natl Acad Sci U S A. 1982 Sep;79(18):5656-60. doi: 10.1073/pnas.79.18.5656.
Resident mouse peritoneal macrophages were labeled with [3H]arachidonic acid and challenged with Sephadex beads coated with immune complexes of IgE and antigen. Arachidonic acid release by the cells was assessed by the quantity or radiolabel recovered from the culture medium. Freshly isolated macrophages responded to IgE immune complexes with a release of [3H]arachidonic acid that was linear for 1-2 hr. The magnitude of the response was dependent on both the number of immune complex-coated beads and on the degree of opsonization of the beads. Under conditions of maximal stimulation, macrophages challenged with IgE immune complex-coated Sephadex released 23 +/- 4.5% of their incorporated radiolabel. This is compared to values of 34.2 +/- 0.5% and 38.1 +/- 3.3% for cultures that received IgG immune complex-coated Sephadex or zymosan, respectively. Macrophages did not release arachidonic acid upon exposure to soluble IgE and antigen given sequentially or simultaneously, and soluble IgE did not inhibit the cells' response to IgE immune complexes. Incubation of macrophages for longer than 3 hr prior to challenge resulted in a selective loss in the cells' ability to respond to IgE immune complexes. After 16 hr of culture, macrophages released only 3.9 +/- 0.3% of their incorporated 3H on exposure to IgE immune complexes; however radiolabel release in response to zymosan (42.0 +/- 0.8%) was identical to that of freshly isolated cells. These data indicate that macrophages are capable of releasing arachidonic acid in response to preformed particulate immune complexes of IgE and antigen. Because Sephadex beads are too large to be interiorized by the cells, this response results from the interaction of the immune complexes with the macrophage plasma membrane.
用[3H]花生四烯酸标记小鼠腹腔巨噬细胞,并使其与包被有IgE和抗原免疫复合物的葡聚糖凝胶珠接触。通过从培养基中回收的放射性标记物的量来评估细胞释放花生四烯酸的情况。新鲜分离的巨噬细胞对IgE免疫复合物有反应,释放[3H]花生四烯酸,在1 - 2小时内呈线性。反应的强度取决于免疫复合物包被珠的数量和珠的调理作用程度。在最大刺激条件下,用IgE免疫复合物包被的葡聚糖凝胶珠刺激的巨噬细胞释放其掺入的放射性标记物的23±4.5%。相比之下,分别接受IgG免疫复合物包被的葡聚糖凝胶珠或酵母聚糖刺激的培养物的释放值为34.2±0.5%和38.1±3.3%。巨噬细胞在依次或同时暴露于可溶性IgE和抗原时不释放花生四烯酸,并且可溶性IgE不抑制细胞对IgE免疫复合物的反应。在刺激前将巨噬细胞孵育超过3小时会导致细胞对IgE免疫复合物反应能力的选择性丧失。培养16小时后,巨噬细胞在暴露于IgE免疫复合物时仅释放其掺入的3H的3.9±0.3%;然而,对酵母聚糖的放射性标记物释放(42.0±0.8%)与新鲜分离的细胞相同。这些数据表明巨噬细胞能够响应预先形成的IgE和抗原颗粒免疫复合物释放花生四烯酸。由于葡聚糖凝胶珠太大而不能被细胞内化,这种反应是由免疫复合物与巨噬细胞质膜的相互作用引起的。