Ishizaka T, Dvorak A M, Conrad D H, Niebyl J R, Marquette J P, Ishizaka K
J Immunol. 1985 Jan;134(1):532-40.
Selective growth of human basophilic granulocytes was obtained in suspension cultures of mononuclear cells from umbilical cord blood. Approximately 50 to 80% of nonadherent cells recovered from 2- to 3-wk-old cultures contained metachromatic granules, and these cells were identified as human basophilic granulocytes by electron microscopy. Histamine content of cultured human basophils was comparable to that in peripheral blood basophils. Cultured basophils bear 2.7 to 3.7 X 10(5) IgE receptors per cell that bind both human IgE and rodent IgE with comparable affinity. Average equilibrium constants of the receptors for human IgE and mouse IgE were 2.56 +/- 0.88 X 10(9) M-1 and 1.85 +/- 0.86 X 10(9) M-1, respectively. The cell-surface component of the IgE receptors on cultured basophils has a m.w. of 64,000. Cultured basophils could be passively sensitized with human IgE and mouse IgE monoclonal antibody, and sensitized basophils released characteristic cytoplasmic granules and both histamine and arachidonate upon challenge with either anti-human IgE or antigen. Incubation of cultured basophils with ionophore A23187 or F-Met-Leu-Phe resulted in histamine release. However, compound 48/80 failed to induce histamine release from the cells.
在脐血单个核细胞的悬浮培养中实现了人嗜碱性粒细胞的选择性生长。从2至3周龄培养物中回收的非贴壁细胞中,约50%至80%含有异染颗粒,通过电子显微镜鉴定这些细胞为人嗜碱性粒细胞。培养的人嗜碱性粒细胞的组胺含量与外周血嗜碱性粒细胞中的相当。培养的嗜碱性粒细胞每个细胞带有2.7至3.7×10⁵个IgE受体,这些受体以相当的亲和力结合人IgE和啮齿动物IgE。人IgE和小鼠IgE受体的平均平衡常数分别为2.56±0.88×10⁹ M⁻¹和1.85±0.86×10⁹ M⁻¹。培养的嗜碱性粒细胞上IgE受体的细胞表面成分的分子量为64,000。培养的嗜碱性粒细胞可用人IgE和小鼠IgE单克隆抗体被动致敏,致敏的嗜碱性粒细胞在受到抗人IgE或抗原攻击时会释放特征性的细胞质颗粒以及组胺和花生四烯酸。用离子载体A23187或F-Met-Leu-Phe孵育培养的嗜碱性粒细胞会导致组胺释放。然而,化合物48/80未能诱导细胞释放组胺。