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Title
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Tracking dye-independent approach to identify and isolate in vitro expanded T cells
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Author
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Abstract
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| T cell proliferation is routinely identified in vitro using tracking dyes or through detecting intracellular upregulation of the nuclear protein, Ki-67. However, labeling with tracking dyes is cumbersome, associated with cellular toxicity, while Ki-67 cannot be used to identify and isolate viable T cells, and both techniques are incompatible with MACS technology. Here, we introduce a simple tool to identify and isolate in vitro T cell expansion that is tracking dye-independent and allows for sorting of viable T cells. We show that CD71, a transferrin receptor, and CD98, a heterodimer glycoprotein involved in both integrin signaling and amino-acid transport, are both highly upregulated on proliferating T cells upon in vitro stimulation, and that CD71 expression is maximal on the more recent progeny T cells, while CD98 upregulation remains stable across different generations of progeny T cells. Moreover, we demonstrate that the upregulation of CD71 and CD98 identifies CFSElow T cells and provides further proof of the antigen-specificity of T cells identified by CD71 and CD98 dual upregulation based on tetramer staining. We further show that CD71 can be used to enrich for in vitro expanding T cells using MACS technology. In conclusion, we show that CD71 and CD98 can be used to identify and isolate expanded T cells following in vitro stimulation and that CD71 is an MACS-compatible alternative to tracking dyes or Ki-67 detection. (c) 2019 International Society for Advancement of Cytometry |
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Language
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English
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Source (journal)
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Cytometry: part A. - New York, 2003, currens
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Publication
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Hoboken
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Wiley
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2019
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ISSN
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1552-4922
[print]
1552-4930
[online]
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DOI
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10.1002/CYTO.A.23867
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Volume/pages
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12 p.
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ISI
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000479401700001
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Pubmed ID
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31356002
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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