Avoiding phagocytosis-related artifact in myeloid derived suppressor cell T-lymphocyte suppression assays

Ruth J. Davis, Christopher Silvin, Clint T. Allen

Abstract

Myeloid-derived suppressor cells (MDSCs) have garnered much attention in recent years as a potential target for altering the immunosuppressive tumor microenvironment in a variety of solid tumor types. The ability to accurately assess the immunosuppressive capacity of MDSCs is fundamental to the development of therapeutic approaches aimed at disabling these immunosuppressive functions. In this article we provide evidence that the use of CD3/28 coated microbeads leads to artefactual T-lymphocyte suppression due to sequestration of beads by MDSCs isolated from the spleens of wild-type mice bearing subcutaneous syngeneic, carcinogen-induced oral cavity carcinomas. Mechanisms of this finding may include early MDSC death and acquisition of phagocytic capacity. These artefactual findings were avoided by eliminating the use of microbeads and instead using plate bound CD3/28 antibody as the T-lymphocyte stimulus. We propose model-specific validation of microbead-based MDSC assays, or use of an alternative stimulation approach such as plate bound CD3/28 antibodies.

Original languageEnglish (US)
Pages (from-to)12-18
Number of pages7
JournalJournal of Immunological Methods
Volume440
DOIs
StatePublished - Jan 1 2017

Fingerprint

Phagocytosis
Artifacts
T-Lymphocytes
Immunosuppressive Agents
Microspheres
Antibodies
Tumor Microenvironment
Carcinogens
Mouth
Cell Death
Spleen
Carcinoma
Neoplasms

Keywords

  • Artifact
  • CD3/28 microbeads
  • MDSC
  • Myeloid-derived suppressor cells
  • Phagocytosis
  • T-cell suppression assay

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Avoiding phagocytosis-related artifact in myeloid derived suppressor cell T-lymphocyte suppression assays. / Davis, Ruth J.; Silvin, Christopher; Allen, Clint T.

In: Journal of Immunological Methods, Vol. 440, 01.01.2017, p. 12-18.

Research output: Contribution to journalReview article

Davis, Ruth J.; Silvin, Christopher; Allen, Clint T. / Avoiding phagocytosis-related artifact in myeloid derived suppressor cell T-lymphocyte suppression assays.

In: Journal of Immunological Methods, Vol. 440, 01.01.2017, p. 12-18.

Research output: Contribution to journalReview article

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AB - Myeloid-derived suppressor cells (MDSCs) have garnered much attention in recent years as a potential target for altering the immunosuppressive tumor microenvironment in a variety of solid tumor types. The ability to accurately assess the immunosuppressive capacity of MDSCs is fundamental to the development of therapeutic approaches aimed at disabling these immunosuppressive functions. In this article we provide evidence that the use of CD3/28 coated microbeads leads to artefactual T-lymphocyte suppression due to sequestration of beads by MDSCs isolated from the spleens of wild-type mice bearing subcutaneous syngeneic, carcinogen-induced oral cavity carcinomas. Mechanisms of this finding may include early MDSC death and acquisition of phagocytic capacity. These artefactual findings were avoided by eliminating the use of microbeads and instead using plate bound CD3/28 antibody as the T-lymphocyte stimulus. We propose model-specific validation of microbead-based MDSC assays, or use of an alternative stimulation approach such as plate bound CD3/28 antibodies.

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