Inagaki N, Kawasaki H, Ueno M, Nagai H, Koda A
Department of Pharmacology, Gifu Pharmaceutical University, Japan.
Life Sci. 1994;54(19):1403-9. doi: 10.1016/0024-3205(94)00595-8.
Rat peritoneal mast cells, which had been sensitized two days earlier by an intraperitoneal injection of rat monoclonal IgE antibodies, were purified by density gradient centrifugation with 60% Percoll (cell purity > 95%). Histamine release from the purified mast cells (PMC) was then compared to that of a non-purified preparation (peritoneal exudate cells; PEC). Both PEC and PMC released similar amounts of histamine upon stimulation with calcium ionophore A23187 and compound 48/80. In contrast, antigen-induced histamine release from PMC was very low compared to that of PEC. PEC released up to 30% of total histamine upon challenge with 1 microgram/ml of antigen, whereas histamine release from PMC was only one third or less than that of PEC. When PEC was suspended in 60% Percoll and treated for a period needed for purification, the reduction of antigen-induced histamine release was negligible. Mast cells purified by centrifugation on a metrizamide gradient released only small amount of histamine similar to Percoll-purified mast cells. Non-mast cells (NMC) recovered from the interface of the 60% Percoll potentiated the antigen-induced histamine release from PMC concentration- and time-dependently. The supernatant of the NMC suspension which was incubated at 37 degrees C for 60 min, however, failed to potentiate histamine release in PMC. We concluded therefore that separation media such as Percoll and metrizamide do not cause the low antigen-induced histamine release in PMC, but that the separation of mast cells from other cells present in the peritoneal cavity itself causes it. Antigen-induced mast cell histamine release is potentiated through a direct interaction between mast cells and NMC, and some cell surface molecules also seem to be involved.
大鼠腹膜肥大细胞在两天前通过腹腔注射大鼠单克隆IgE抗体进行致敏,然后用60%的 Percoll通过密度梯度离心进行纯化(细胞纯度>95%)。然后将纯化的肥大细胞(PMC)的组胺释放与未纯化的制剂(腹膜渗出细胞;PEC)的组胺释放进行比较。在用钙离子载体A23187和化合物48/80刺激时,PEC和PMC释放的组胺量相似。相比之下,与PEC相比,抗原诱导的PMC组胺释放非常低。在用1微克/毫升抗原攻击时,PEC释放的组胺可达总组胺的30%,而PMC的组胺释放仅为PEC的三分之一或更少。当PEC悬浮在60%的 Percoll中并进行纯化所需的处理时,抗原诱导的组胺释放的减少可以忽略不计。通过在甲泛葡胺梯度上离心纯化的肥大细胞释放的组胺量很少,类似于用 Percoll纯化的肥大细胞。从60% Percoll的界面回收的非肥大细胞(NMC)浓度和时间依赖性地增强了抗原诱导的PMC组胺释放。然而,在37℃孵育60分钟的NMC悬浮液的上清液未能增强PMC中的组胺释放。因此,我们得出结论,诸如 Percoll和甲泛葡胺等分离介质不会导致PMC中抗原诱导的组胺释放低,而是腹膜腔中存在的肥大细胞与其他细胞的分离导致了这种情况。抗原诱导的肥大细胞组胺释放通过肥大细胞与NMC之间的直接相互作用而增强,并且一些细胞表面分子似乎也参与其中。