Tracking Dye-Independent Approach to Identify and Isolate In Vitro Expanded T Cells

George Elias, Benson Ogunjimi, Viggo Van Tendeloo

Onderzoeksoutput: Articlepeer review

3 Citaten (Scopus)

Samenvatting

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. © 2019 International Society for Advancement of Cytometry.

Originele taal-2English
Pagina's (van-tot)1096-1107
Aantal pagina's12
TijdschriftCytometry. Part A
Volume95
Nummer van het tijdschrift10
DOI's
StatusPublished - okt 2019

Bibliografische nota

© 2019 International Society for Advancement of Cytometry.

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