A novel method to determine the engulfment of apoptotic cells by macrophages using pHrodo succinimidyl ester.
A novel method to determine the engulfment of apoptotic cells by macrophages using pHrodo succinimidyl ester.
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DOI：
10.1016/j.jim.2008.11.019
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发表时间：
2009-03-15
影响因子：
2.2
通讯作者：
Wang, Ping
中科院分区：
文献类型：
--
作者：
Miksa, Michael;Kornura, Hidefumi;Wu, Rongqian;Shah, Kavin G.;Wang, Ping
最近，凋亡细胞吞噬作用引起了相当大的兴趣，特别是由于其复杂的分子机制和对凋亡细胞清除能力不足的负面免疫学影响。需要简单和可靠的方法来清楚地确定凋亡细胞的内在化。用pH敏感的荧光染料PHRODO琥珀酰亚胺酯(SE)标记，由于吞噬后发光增加，可以检测到吞噬的凋亡细胞。这是通过流式细胞仪研究吞噬作用的一个有价值的工具。我们设计了一种体外实验，使用凋亡的pHrodo标记的淋巴细胞作为猎物，并使用抗CD11b标记的组织巨噬细胞。为了证明它检测内化的凋亡淋巴细胞的有效性，我们使用了已知具有吞噬能力受损的MFGE8-/-巨噬细胞。重组MFGE8刺激后，脾巨噬细胞对凋亡淋巴细胞的摄取加快和增强，而腹膜巨噬细胞则能补偿这种延迟摄取。这种新的检测方法是一种快速而可靠的方法来评估凋亡细胞的内在化。
Apoptotic cell phagocytosis has recently raised considerable interest, particularly due to its intricate molecular mechanisms and negative immunologic impact of incompetent clearance of apoptotic cells. There is a need for simple and reliable methods to clearly determine the internalization of apoptotic cells. Labeling with pHrodo succinimidyl ester (SE), a pH-sensitive fluorescent dye, makes engulfed apoptotic cells detectable due to the increased post-phagocytic light emission. This is a valuable tool for phagocytosis studies via FACS. We designed an ex vivo assay, using apoptotic pHrodo-labeled lymphocytes as prey and anti-CD11b-labeled tissue macrophages. To demonstrate its validity of detecting internalized apoptotic lymphocytes, we used MFGE8-/- macrophages, known to have impaired phagocytic ability. Uptake of apoptotic lymphocytes was accelerated and enhanced in splenic macrophages after stimulation with recombinant MFGE8, while peritoneal macrophages were able to compensate for the delayed uptake. This novel assay is a quick and reliable method to evaluate the internalization of apoptotic cells.
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影响因子：
56.9
作者：
Blander, JM;Medzhitov, R
通讯作者：
Medzhitov, R
影响因子：
37.8
作者：
Ait-Oufella, Hafid;Kinugawa, Kiyoka;Mallat, Ziad
通讯作者：
Mallat, Ziad
影响因子：
56.9
作者：
Hanayama, R;Tanaka, M;Nagata, S
通讯作者：
Nagata, S
DOI：
10.1084/jem.20040342
发表时间：
2004-08-16
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Asano K;Miwa M;Miwa K;Hanayama R;Nagase H;Nagata S;Tanaka M
通讯作者：
Tanaka M
乳脂肪球-EGF因子8/乳粘素在小鼠肠上皮的维持和修复中发挥着至关重要的作用
影响因子：
15.9
作者：
Bu, Heng-Fu;Zuo, Xiu-Li;Tan, Xiao-Di
通讯作者：
Tan, Xiao-Di