Aberrant capping of membrane proteins on neutrophils from patients with leukocyte adhesion deficiency.
Aberrant capping of membrane proteins on neutrophils from patients with leukocyte adhesion deficiency.
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DOI：
10.1182/blood.v83.6.1650.1650
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发表时间：
1994-03
期刊：
影响因子：
20.3
通讯作者：
A. Kindzelskii;W. Xue;R. Todd;L. Boxer;H. Petty
中科院分区：
文献类型：
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作者：
A. Kindzelskii;W. Xue;R. Todd;L. Boxer;H. Petty
在白细胞黏附缺陷(LAD)患者的中性粒细胞中发现了几种功能缺陷：MO1、LFA-1和p150、95。为了更好地了解LAD中性粒细胞的功能缺陷，我们进行了封顶实验。将纯化的正常或LAD中性粒细胞分别标记抗尿激酶型纤溶酶原激活剂受体(UPAR)、抗Fc-γRIII(CD16)、抗MO5、抗CD14抗体的刀豆蛋白A(ConA)或F(ab‘)2片段。用抗尿Fab抗血清的第二步F(ab‘)2片段封盖F(ab’)2标记的细胞。细胞在37℃下封顶30分钟，然后在荧光显微镜下观察。到目前为止，LAD中性粒细胞被发现在封盖方面存在缺陷，但没有对所有测试的试剂进行聚集。正常中性粒细胞Con A、Fc-γRIII、尿激酶受体、CD14和MO5封闭率分别为52%、67%、70%、25%和%，而LAD中性粒细胞分别为10%、5%、2%、3%和1%。加入10(-5)m ol/L秋水仙碱或10(-7)m o l/L FMLP均不能增加LAD或正常中性粒细胞这一组膜成分的封闭量。由于封顶需要膜到胞浆的通讯和完整的微丝连接，我们认为白细胞粘附素可能在促进膜成分的重新分布方面发挥广泛的作用，包括与黏附相关的受体，如Fc-Gamma RIII和尿激酶受体。
Several functional defects have been found in neutrophils from leukocyte adhesion deficiency (LAD) patients who fail to express the CD11/CD18 leukoadhesins: Mo1, LFA-1, and p150,95. To better understand the functional defects of LAD neutrophils, we have performed capping experiments. Purified normal or LAD neutrophils were labeled with fluorochrome-conjugated concanavalin A (Con A) or F(ab')2 fragments of antiurokinase-type plasminogen activator receptor (uPAR), anti-Fc gamma RIII (CD16), anti-Mo5, and anti-CD14 antibodies. F(ab')2-labeled cells were capped using a second-step F(ab')2 fragment of an antimurine Fab antiserum. Cells were capped for 30 minutes at 37 degrees C, then observed by fluorescence microscopy. LAD neutrophils were found to be deficient in capping, but not clustering of all of the reagents tested to date. The percent of cells exhibiting capping of Con A, Fc gamma RIII, urokinase receptor, CD14, and Mo5 were 52%, 67%, 70%, 25%, and 64% for normal neutrophils but were only 10%, 5%, 2%, 3%, and 1%, respectively, for LAD neutrophils. Capping of this panel of membrane components in LAD or normal neutrophils was not augmented by the addition of either 10(-5) mol/L colchicine or 10(-7) mol/L FMLP. Because capping requires membrane-to-cytosol communication and an intact microfilament linkage, we suggest that leukoadhesins may play a broad role in promoting the redistribution of membrane components including adherence-related receptors such as Fc gamma RIII and the urokinase receptor.