Fc (IgG) receptor distributions in homogeneous and heterogeneous cell populations by flow microfluorometry.
Fc (IgG) receptor distributions in homogeneous and heterogeneous cell populations by flow microfluorometry.
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通过流式显微荧光测定法观察同质和异质细胞群中的 Fc (IgG) 受体分布。
DOI：
10.1073/pnas.78.1.519
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发表时间：
1981
影响因子：
11.1
通讯作者：
D. Segal
中科院分区：
文献类型：
--
作者：
J. A. Titus;S. Sharrow;J. Connolly;D. Segal
流动显微荧光法已被开发用于定量单个细胞上的Fc受体的数量。将细胞在0 ℃下用放射性标记的亲和交联兔IgG二聚体平衡，洗涤，并用针对兔IgG的荧光抗体处理。使用荧光激活细胞分选仪分析染色细胞的荧光发射，并使用γ计数器分析结合的二聚体分子。将荧光发射与结合至细胞的二聚体分子的数量相关的标准曲线用于确定P388 D1细胞、人外周血淋巴细胞和正常小鼠脾细胞上的Fc受体分布。基本上所有的P388 D1细胞都带有Fc受体，以倾斜的高斯分布分布，每个细胞有2 × 10（5）个受体。人外周血淋巴细胞和小鼠脾细胞含有阳性和阴性亚群。来自不同供体的人淋巴细胞中阳性细胞的百分比范围为50 - 25;这些细胞的受体分布是对称的，并且在所有供体中在形状和平均受体密度方面相似（每个细胞4.2 × 10（4）个受体）。小鼠脾细胞含有55%的阳性细胞，受体密度呈非对称异质分布。这些细胞的峰值为每个细胞1至2 × 10（4）受体，但大量细胞的受体密度高出10至20倍。
A flow microfluorometric method has been developed for quantitating the numbers of Fc receptors on individual cells. The cells were equilibrated at 0 degrees C with radiolabeled, affinity-crosslinked rabbit IgG dimers, washed, and treated with fluorescent antibodies against rabbit IgG. The stained cells were analyzed for fluorescence emission by using a fluorescence-activated cell sorter and for bound dimer molecules by using a gamma counter. Standard curves relating fluorescence emission to numbers of dimer molecules bound to cells were used to determine Fc receptor distributions on P388D1 cells, human peripheral blood lymphocytes, and normal mouse spleen cells. Essentially all of the P388D1 cells bore Fc receptors, distributed in a skewed Gaussian profile having a peak at 2 X 10(5) receptors per cell. Human peripheral blood lymphocytes and mouse spleen cells contained positive and negative subpopulations. The percentage of positive cells in human lymphocytes from different donors ranged from 50 to 25; the receptor distributions of these cells were symmetrical and similar in all donors in shape and average receptor density (4.2 X 10(4) receptors per cell). Mouse spleen cells contained 55% positive cells with nonsymmetrical heterogeneous distributions of receptor densities. These cells peaked at 1 to 2 X 10(4) receptors per cell, but significant numbers of cells had receptor densities 10- to 20-fold greater.