{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T17:38:32Z","timestamp":1776533912208,"version":"3.51.2"},"reference-count":49,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T00:00:00Z","timestamp":1697155200000},"content-version":"vor","delay-in-days":12,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2023,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Trogocytosis is a process in which receptors on acceptor cells remove and internalize cognate ligands from donor cells. Trogocytosis has a profound and negative impact on mAb\u2010based cancer immunotherapy, as seen in the treatment of chronic lymphocytic leukemia (CLL) with CD20 mAbs, such as rituximab (RTX) and ofatumumab (OFA). Our clinical observations of RTX\/OFA\u2010mediated loss of the CD20 target from circulating CLL cells have been replicated in our in vitro studies. Here we describe flow cytometry and fluorescence microscopy experiments, which demonstrate that acceptor cells, such as monocytes\/macrophages that express Fc\u03b3R, remove and internalize both antigen and donor cell\u2010bound cognate IgG mAbs for several different mAb\u2010donor cell pairs. Fluorescent mAbs and portions of the plasma cell membrane are transferred from donor cells to acceptor cells, which include the THP\u20101 monocytic cell line as well as freshly isolated monocytes. We describe rigorous controls to validate the reactions and eliminate dissociation or internalization as alternative mechanisms. Trogocytosis is likely to contribute to neutropenia, thrombocytopenia, and liver damage associated with use of antibody\u2010drug conjugates. The methods we have described should allow for examination of strategies focused on blocking trogocytosis and its adverse effects. \u00a9 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Trogocytosis of mAb\u2010opsonized donor cells mediated by adherent THP\u20101 cells<\/jats:p><jats:p><jats:bold>Alternate Protocol<\/jats:bold>: Application of fluorescence microscopy to examine THP\u20101 cell\u2010mediated trogocytosis<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: Alexa labeling of mAbs and determination of F\/P ratios<\/jats:p><jats:p><jats:bold>Support Protocol 2<\/jats:bold>: Standard washing procedure<\/jats:p><jats:p><jats:bold>Support Protocol 3<\/jats:bold>: Labeling and opsonization of cells<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Trogocytosis mediated by human monocytes as acceptor cells<\/jats:p><jats:p><jats:bold>Support Protocol 4<\/jats:bold>: Isolation of human monocytes<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Trogocytosis mediated by THP\u20101 cells in solution<\/jats:p><jats:p><jats:bold>Support Protocol 5<\/jats:bold>: Retinoic acid treatment of THP\u20101 cells<\/jats:p><jats:p><jats:bold>Support Protocol 6<\/jats:bold>: Culturing of SCC\u201025, BT\u2010474, MOLT\u20104 and THP\u20101 cell lines<\/jats:p>","DOI":"10.1002\/cpz1.897","type":"journal-article","created":{"date-parts":[[2023,10,13]],"date-time":"2023-10-13T12:14:57Z","timestamp":1697199297000},"source":"Crossref","is-referenced-by-count":3,"title":["Measurement of Trogocytosis: Quantitative Analyses Validated with Rigorous Controls"],"prefix":"10.1002","volume":"3","author":[{"given":"Ronald P.","family":"Taylor","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics University of Virginia School of Medicine  Charlottesville Virginia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Margaret A.","family":"Lindorfer","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Genetics University of Virginia School of Medicine  Charlottesville Virginia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,10,13]]},"reference":[{"key":"e_1_2_15_2_1","doi-asserted-by":"publisher","DOI":"10.1681\/ASN.2006101125"},{"key":"e_1_2_15_3_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI107769"},{"key":"e_1_2_15_4_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI107325"},{"key":"e_1_2_15_5_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.176.4.2600"},{"key":"e_1_2_15_6_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.181.4.2916"},{"key":"e_1_2_15_7_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.181.11.8120"},{"key":"e_1_2_15_8_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1101189"},{"key":"e_1_2_15_9_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.11036938845"},{"key":"e_1_2_15_10_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI110764"},{"key":"e_1_2_15_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0090\u20101229(87)90167\u2010X"},{"key":"e_1_2_15_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/1529\u20100131(200010)43:10<2265::AID\u2010ANR14>3.0.CO;2\u2010J"},{"key":"e_1_2_15_13_1","doi-asserted-by":"publisher","DOI":"10.1006\/clim.2002.5266"},{"key":"e_1_2_15_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_15_15_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.271.5247.348"},{"key":"e_1_2_15_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/eji.1830220602"},{"key":"e_1_2_15_17_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.139.1.180"},{"key":"e_1_2_15_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/0090\u20101229(89)90212\u20102"},{"key":"e_1_2_15_19_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.18.1.393"},{"key":"e_1_2_15_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im701s858892"},{"key":"e_1_2_15_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.imlet.2004.07.007"},{"key":"e_1_2_15_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/ki.1987.81"},{"key":"e_1_2_15_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni0903\u2010815"},{"key":"e_1_2_15_24_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.172.5.3280"},{"key":"e_1_2_15_25_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI114259"},{"key":"e_1_2_15_26_1","doi-asserted-by":"publisher","DOI":"10.1158\/1078\u20100432.CCR\u201017\u201020273583"},{"key":"e_1_2_15_27_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.160.10.5088"},{"key":"e_1_2_15_28_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.170.7.3671"},{"key":"e_1_2_15_29_1","doi-asserted-by":"publisher","DOI":"10.3390\/antib110310458474"},{"key":"e_1_2_15_30_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.114.4.1166"},{"key":"e_1_2_15_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1521\u20104141(199905)29:05<1581::AID\u2010IMMU1581>3.0.CO;2\u2010V"},{"key":"e_1_2_15_32_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.118.3077.733"},{"key":"e_1_2_15_33_1","doi-asserted-by":"publisher","DOI":"10.1177\/003591575604900122"},{"key":"e_1_2_15_34_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood\u20102016\u201004\u20107095198891"},{"key":"e_1_2_15_35_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.135.3.2005"},{"key":"e_1_2_15_36_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI106847"},{"key":"e_1_2_15_37_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI106846"},{"key":"e_1_2_15_38_1","doi-asserted-by":"publisher","DOI":"10.1006\/clin.1996.4286"},{"key":"e_1_2_15_39_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.136.10.3785"},{"key":"e_1_2_15_40_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.134.4.2550"},{"key":"e_1_2_15_41_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood\u20102014\u201010\u2010569244"},{"key":"e_1_2_15_42_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.159.8.4035"},{"key":"e_1_2_15_43_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.148.8.2462"},{"key":"e_1_2_15_44_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.158.2.842"},{"key":"e_1_2_15_45_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.20220093828893"},{"key":"e_1_2_15_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2006.02.032"},{"key":"e_1_2_15_47_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.173.4.22978696"},{"key":"e_1_2_15_48_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.177.10.7435"},{"key":"e_1_2_15_49_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.237377527"},{"key":"e_1_2_15_50_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.242657528"}],"container-title":["Current 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