Vally H, Savigni D L, Turner K J
Department of Microbiology, University of Western Australia, Nedlands.
Immunol Cell Biol. 1995 Jun;73(3):212-9. doi: 10.1038/icb.1995.35.
Mast cells were purified by discontinuous density gradient centrifugation to greater than 90% and subsequently exposed to lactic acid in order to dissociate endogenous IgE. Unfortunately no removal of this IgE was achieved, despite previous reports of lactic acid exposure dissociating IgE from human basophils. Only a minimal level of endogenous IgE was found on the surface of mast cells, however, which allowed investigations of IgE binding to proceed without the initial removal of this endogenous IgE. Conditions for binding of normal rat IgE and myeloma IgE were established while efforts to bind human IgE to purified rat mast cells were unsuccessful. Our results suggest that while purified rat mast cells have the potential to play an important role in the study of the IgE requirement of various stimuli including histamine-releasing factor, this role is limited at present.
肥大细胞通过不连续密度梯度离心法纯化至纯度超过90%，随后暴露于乳酸中以解离内源性IgE。遗憾的是，尽管先前有报道称乳酸暴露可使IgE从人嗜碱性粒细胞上解离，但并未实现对内源性IgE的去除。然而，在肥大细胞表面仅发现了极低水平的内源性IgE，这使得在不预先去除这种内源性IgE的情况下就能够进行IgE结合的研究。确定了正常大鼠IgE和骨髓瘤IgE的结合条件，而将人IgE结合到纯化的大鼠肥大细胞上的尝试未成功。我们的结果表明，虽然纯化的大鼠肥大细胞在包括组胺释放因子在内的各种刺激的IgE需求研究中有可能发挥重要作用，但目前这一作用有限。
