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Plos One : Mechanism of Fcc Receptor-mediated Trogocytosis- Based False-positive Results in Flow Cytometry, Volume 7

By Hoshino, Yoshihiko

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Book Id: WPLBN0003940492
Format Type: PDF eBook :
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Reproduction Date: 2015

Title: Plos One : Mechanism of Fcc Receptor-mediated Trogocytosis- Based False-positive Results in Flow Cytometry, Volume 7  
Author: Hoshino, Yoshihiko
Volume: Volume 7
Language: English
Subject: Journals, Science, Medical Science
Collections: Periodicals: Journal and Magazine Collection (Contemporary)
Historic
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Publisher: Plos

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Hoshino, Y. (n.d.). Plos One : Mechanism of Fcc Receptor-mediated Trogocytosis- Based False-positive Results in Flow Cytometry, Volume 7. Retrieved from http://netlibrary.net/


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Description : 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 Fcc receptor (FccR)-mediated trogocytosis. In the present study, similar FccR-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 FccR expression in peripheral blood, could perform FccR-mediated trogocytosis. Since the capacity of FccR-mediated trogocytosis was different among blood samples, identification of factors that could regulate the occurrence of FccR-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 FccR-mediated trogocytosis using granulocytes. Investigation with this model determined the serum factors as heatlabile 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 FccR-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 FccR-mediated trogocytosis. Moreover, the inhibition rates were significantly higher in HAMAhigh serum than HAMAlow serum. The collective findings suggested the involvement of heterophilic antibodies such as HAMA in the mechanism of false-positive results in FCM due to FccR-mediated trogocytosis.

 

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