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Mechanism of Fcγ Receptor-Mediated Trogocytosis-Based False-Positive Results in Flow Cytometry
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Title: | Mechanism of Fcγ Receptor-Mediated Trogocytosis-Based False-Positive Results in Flow Cytometry |
Authors: | Masuda, Sakiko Browse this author | Iwasaki, Sari Browse this author →KAKEN DB | Tomaru, Utano Browse this author →KAKEN DB | Sato, Juri Browse this author | Kawakami, Ai Browse this author | Ichijo, Kana Browse this author | Sogo, Sayuri Browse this author | Baba, Tomohisa Browse this author | Katsumata, Kazuaki Browse this author | Kasahara, Masanori Browse this author →KAKEN DB | Ishizu, Akihiro Browse this author →KAKEN DB |
Issue Date: | 27-Dec-2012 |
Publisher: | Public Library of Science |
Journal Title: | PLoS One |
Volume: | 7 |
Issue: | 12 |
Start Page: | e52918 |
Publisher DOI: | 10.1371/journal.pone.0052918 |
Abstract: | The whole blood erythrocyte lysis method is the most common protocol of sample preparation for flow cytometry (FCM). Although this method has many virtues, our recent study has demonstrated false-positive results when surface markers of monocytes were examined by this method due to the phenomenon called Fcγ receptor (FcγR)-mediated trogocytosis. In the present study, similar FcγR-mediated trogocytosis-based false-positive results have been demonstrated when granulocytes were focused on instead of monocytes. These findings indicated that not only monocytes but also granulocytes, the largest population with FcγR expression in peripheral blood, could perform FcγR-mediated trogocytosis. Since the capacity of FcγR-mediated trogocytosis was different among blood samples, identification of factors that could regulate the occurrence of FcγR-mediated trogocytosis should be important for the quality control of FCM. Our studies have suggested that such factors are present in the serum. In order to identify the serum factors, we employed the in vitro model of FcγR-mediated trogocytosis using granulocytes. Investigation with this model determined the serum factors as heat-labile molecules with molecular weight of more than 100 kDa. Complements in the classical pathway were initially assumed as candidates; however, the C1 inhibitor did not yield an obvious influence on FcγR-mediated trogocytosis. On the other hand, although immunoglobulin ought to be resistant to heat inactivation, the inhibitor of human anti-mouse antibodies (HAMA) effectively blocked FcγR-mediated trogocytosis. Moreover, the inhibition rates were significantly higher in HAMA^[high] serum than HAMA^[low] serum. The collective findings suggested the involvement of heterophilic antibodies such as HAMA in the mechanism of false-positive results in FCM due to FcγR-mediated trogocytosis. |
Rights: | http://creativecommons.org/licenses/by/3.0/ |
Type: | article |
URI: | http://hdl.handle.net/2115/51771 |
Appears in Collections: | 保健科学院・保健科学研究院 (Graduate School of Health Sciences / Faculty of Health Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 石津 明洋
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