The Value of a Rapid Test of Human Regulatory T Cell Function Needs to be Revised

Frontiers in Immunology
Desiree J WenderingMichael Schmueck-Henneresse

Abstract

CD4+CD25+FoxP3+ human regulatory TCELLS (TREG) are promising candidates for reshaping undesired immunity/inflammation by adoptive cell transfer, yet their application is strongly dependent on robust assays testing their functionality. Several studies along with first clinical data indicate TREG to be auspicious to use for future cell therapies, e.g., to induce tolerance after solid organ transplantation. To this end, TREG suppressive capacity has to be thoroughly evaluated prior to any therapeutic application. A 7 h-protocol for the assessment of TREG function by suppression of the early activation markers CD154 and CD69 on CD4+CD25- responder TCELLS (TRESP) upon polyclonal stimulation via αCD3/28-coated activating microbeads has previously been published. Even though this assay has since been applied by various groups, the protocol comes with a critical pitfall, which is yet not corrected by the journal of its original publication. Our results demonstrate that the observed decrease in activation marker frequency on TRESP is due to competition for αCD3/28-coated microbeads as opposed to a TREG-attributable effect and therefore the protocol cannot further be used as a diagnostic test to assess suppressive TREG function.

References

Dec 29, 2007·Immunological Investigations·Todd M BruskoMark A Atkinson
May 31, 2008·Cell·Shimon SakaguchiMasahiro Ono
Jul 26, 2011·Journal of Immunological Methods·Joyce J RuitenbergSmita A Ghanekar
Jan 29, 2014·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·S Landwehr-KenzelP Reinke
Nov 5, 2014·Immunopharmacology and Immunotoxicology·Anders Elm PedersenMonika Gad
Apr 14, 2016·Mediators of Inflammation·Johanna RuhnauAlexander Dressel

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Citations

Jan 27, 2020·Trends in Biotechnology·Enrico FritscheMohamed Abou-El-Enein

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Methods Mentioned

BETA
fluorescence-activated cell sorting
flow cytometry
FCS

Software Mentioned

nparLD
TreeStar
R
GraphPad Prism
FlowJo

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