PROPERTIES OF HUMAN BASOPHILS ISOLATED BY FLUORESCENCE-ACTIVATED CELL SORTING
PROPERTIES OF HUMAN BASOPHILS ISOLATED BY FLUORESCENCE-ACTIVATED CELL SORTING
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DOI：
10.1016/0091-6749(88)90019-x
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发表时间：
1988-09-01
影响因子：
14.2
通讯作者：
NEIJENS, HJ
中科院分区：
文献类型：
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作者：
DEGENHART, HJ;VANDERMAAREL, JWM;NEIJENS, HJ
本研究的目的有两个：（1）考虑荧光激活细胞分选（FACS）对从小血容量中分离嗜碱性粒细胞的适用性，（2）比较从哮喘儿童中获得的嗜碱性粒细胞与健康儿童中获得的嗜碱性粒细胞。采用FACS，制备了纯度为84%（范围：75%-95%）、回收率为20%（范围：15%-30%）的嗜碱性粒细胞混悬液。纯化的嗜碱性粒细胞的总组胺含量为1.6 ± 1.5。0.12 pg/细胞，与在“总”白细胞悬浮液中观察到的总组胺含量没有显著差异（1.4 ± 0.001）。0.07 pg/嗜碱性粒细胞）。对于IgE受体介导的组胺释放也是如此（29 . ±. 4%对27 .+-. 4%）和离子载体A23187诱导的组胺释放（41 . ±. 6%对51 .+-. 9%）。来自哮喘受试者的分选的嗜碱性粒细胞在IgE受体活化后释放更多的组胺（0.67 ± 0.99）。0.09 pg/细胞）比来自健康儿童的嗜碱性粒细胞（0.40 ± 0.50 μ g/细胞）高。0.04 pg/细胞; p < 0.02）。以释放百分比表示，没有观察到显著差异（37 ± 0.5%）。3.2%对30 .+-. 2.7%）。离子载体A23187诱导的组胺释放在哮喘受试者和对照受试者之间没有显著差异，也没有以皮克/细胞表示（1.21 ± 0.01）。0.17 pg/细胞对1.02 ±. 0.11 pg/细胞），也不表示为释放百分比（66 ± 0.5pg/细胞）。4.4%与74 .+-. 3.2%）。由Ca 2+暴露和离子载体A23187暴露诱导的IgE受体介导的膜电位变化证明哮喘受试者和健康儿童之间没有显著差异：12 . ±. 1.6 mV与14 . ±. 1.0在Ca 2+暴露后11 . ±. 18 mV vs 12 . ±. 1.4 mV的离子载体A23187的存在下。流式细胞仪非常适合于制备高纯度的嗜碱性粒细胞悬浮液，没有显着的选择偏差或细胞损伤。在IgE刺激下，哮喘患者的纯化嗜碱性粒细胞比健康儿童的嗜碱性粒细胞释放更多的组胺。在离子载体A23187诱导的组胺释放中没有这种差异。
The aim of the current study was twofold: (1) to consider the applicability of fluorescence-activated cell sorting (FACS) to basophil isolation from small blood volumes and (2) to compare basophils obtained from children with asthma to basophils from healthy children. With FACS, basophil suspensions were prepared with a purity of 84% (range, 75% to 95%) and a recovery of 20% (range, 15% to 30%). The purified basophils had a total histamine content of 1.6 .+-. 0.12 pg per cell, not differing significantly from total histamine content observed in "total" leukocyte suspensions (1.4 .+-. 0.07 pg per basophil). The same was true for IgE receptor-mediated histamine release (29 .+-. 4% versus 27 .+-. 4%) and for ionophore A23187-induced histamine release (41 .+-. 6% versus 51 .+-. 9%). Sorted basophils from subjects with asthma released more histamine after IgE receptor activation (0.67 .+-. 0.09 pg per cell) than basophils from healthy children (0.40 .+-. 0.04 pg per cell; p < 0.02). Expressed as percent release, no significant difference was observed (37 .+-. 3.2% versus 30 .+-. 2.7%). Ionophore A23187-induced histamine release did not differ significantly between subjects with asthma and control subjects, neither expressed as picograms per cell (1.21 .+-. 0.17 pg per cell versus 1.02 .+-. 0.11 pg per cell) nor expressed as percent release (66 .+-. 4.4% versus 74 .+-. 3.2%). The IgE receptor-mediated change in membrane potential induced by Ca2+ exposure and by ionoiphore A23187 exposure demonstrated no significant differences between subjects with asthma and healthy children: 12 .+-. 1.6 mV versus 14 .+-. 1.0 mV after Ca2+ exposure and 11 .+-. 18 mV versus 12 .+-. 1.4 mV in the presence of ionophore A23187. FACS is well suited for the preparation of high-purity basophil suspensions without significant selection bias or cell damage. In response to IgE stiulation, purified basophils from subjects with asthma release more histaminethan basophils from healthy children. No such differences could be demonstrated in ionophore A23187-induced histamne release.