Influence of acridine tracer dyes on neutrophil function.
Influence of acridine tracer dyes on neutrophil function.
复制标题
吖啶示踪染料对中性粒细胞功能的影响。
DOI：
10.1002/jlb.56.4.464
复制
发表时间：
1994
影响因子：
5.5
通讯作者：
Arfors,KE
中科院分区：
文献类型：
--
作者：
Hansell,P;Berger,E;Chambers,JD;Arfors,KE
研究了体内显微镜研究中常用的两种荧光染料--丫啶橙(AO)和丫啶红(AR)对佛波酯(PMA)或甲酰肽(FMLP)刺激的人中性粒细胞黏附牛血清白蛋白基质和产生超氧阴离子(SOX)能力的影响。未标记刺激的人中性粒细胞与基质的粘附率分别为36±9%(PMA，10-7M)和11±7%(fMLP，10-7M)。这种黏附是CD18依赖的，当加入抗CD18抗体IB4时，黏附分别减少了98%和92%。体外染料标记30min后，对刺激的人中性粒细胞黏附的抑制作用呈剂量依赖性，EC50值分别为70μg/ml(AR)和145μg/ml(AO)。PMA刺激的中性粒细胞产生的SOx在100μg/mlAR和AO时不受影响，但在较高剂量时减少40-60%。体内和体外用100μg/mlAR标记的兔中性粒细胞SOX生成量分别降低41%和61%。研究表明，中性粒细胞的功能，就使用CD11/CD18整合素黏附于BSA基质并在刺激下产生SOX的能力而言，取决于剂量和荧光染料的选择。在研究PMN功能时，应注意在高浓度下使用这些化合物。J.Leukoc。比奥尔。56：464-468；1994。
A study was performed to elucidate the effect of two commonly used fluorescent dyes in in vivo microscopic studies, acridine orange (AO) and acridine red (AR), on the ability of phorbol myristate acetate (PMA)-or formyl peptide (fMLP)-stimulated human neutrophils to adhere to a bovine serum albumin matrix and to generate superoxide anions (SOX). Unlabeled stimulated human neutrophils showed 36 ± 9% (PMA, 10-7M) and 11 ± 7% (fMLP, 10-7M) adherence to the matrix. This adhesion was CD18 dependent as evidenced by 98% and 92% reduction, respectively, when the anti-CD18 antibody IB4 was included. A dose-dependent inhibition of stimulated human neutrophil adhesion was evident after 30 min of in vitro dye labeling and the EC50was approximately 70μg/ml (AR) and 145μg/ml (AO). SOX generation by PMA-stimulated neutrophils was unaffected up to 100μg/ml AR and AO but was reduced by 40–60% at higher doses. Rabbit neutrophils labeled in vivo or in vitro with 100μg/ml AR exhibited 41% and 61% lower SOX generation, respectively. The study indicates that neutrophil function, in terms of ability to adhere to a BSA matrix using CD11/CD18 integrins and to generate SOX upon stimulation, is reduced depending on the dose and choice of fluorescent dye. Caution should be exercised when using these compounds at high concentrations in studies of PMN function. J. Leukoc. Biol. 56: 464–468; 1994.