Lett-Brown M A, Robinson L, Juneja H S, Grant J A
Department of Internal Medicine, University of Texas Medical Branch, Galveston 77550.
J Immunol Methods. 1989 Feb 24;117(2):163-7. doi: 10.1016/0022-1759(89)90136-1.
Although usually the least prevalent blood leukocyte, the basophil can release potent soluble factors in response to multiple triggers. We purified basophils from normal volunteers by means of isopycnic centrifugation and affinity binding of mononuclear cells. The majority of the basophils from most subjects were recovered in a band formed between Percoll layers with densities of 1.070 and 1.080; at this stage basophils represented a mean of 22% of total leukocytes. These cells were reacted with monoclonal antibodies to T (OKT-11) and B (anti-HLA-DR) lymphocytes; B and T cells were removed by adsorption to insoluble antibodies against mouse immunoglobulin resulting in a mean purity of 75% basophils with a yield of 54%. These highly enriched basophils resembled unpurified basophils in terms of (1) intracellular histamine content, (2) spontaneous release of histamine in buffer, and (3) percentage of histamine released by anti-IgE. These findings suggest that the techniques used to purify the basophils do not affect the functional integrity of human basophils.
尽管嗜碱性粒细胞通常是血液中最不常见的白细胞，但它能对多种刺激做出反应，释放出强效的可溶性因子。我们通过等密度离心和单核细胞的亲和结合从正常志愿者中纯化嗜碱性粒细胞。大多数受试者的嗜碱性粒细胞大部分在密度为1.070至1.080的Percoll层之间形成的条带中回收；在此阶段，嗜碱性粒细胞平均占白细胞总数的22%。这些细胞与针对T（OKT-11）和B（抗HLA-DR）淋巴细胞的单克隆抗体反应；通过吸附到抗小鼠免疫球蛋白的不溶性抗体上去除B细胞和T细胞，最终嗜碱性粒细胞的平均纯度为75%，产率为54%。这些高度富集的嗜碱性粒细胞在以下方面与未纯化的嗜碱性粒细胞相似：（1）细胞内组胺含量，（2）缓冲液中组胺的自发释放，以及（3）抗IgE释放的组胺百分比。这些发现表明，用于纯化嗜碱性粒细胞的技术不会影响人嗜碱性粒细胞的功能完整性。
