Cytotechnology

, Volume 70, Issue 1, pp 1–11 | Cite as

Development of an 8-color antibody panel for functional phenotyping of human CD8+ cytotoxic T cells from peripheral blood mononuclear cells

  • Tara Patel
  • Amy Cunningham
  • Martha Holland
  • John Daley
  • Suzan Lazo
  • F. Stephen Hodi
  • Mariano Severgnini
Methods Paper
  • 240 Downloads

Abstract

The study of CD8 positive cells in peripheral blood has become an essential part of research in the field of cancer immunotherapies, vaccine development, inflammation, autoimmune disease, etc. In this study, an 8-color flow cytometry panel, containing lineage and functional markers, was developed for the identification of CD8+ cytotoxic T cells in previously cryopreserved peripheral blood mononuclear cells from healthy human donors. By studying functional markers in naïve and CD3/CD28 activated T cells we demonstrate that the panel is capable of detecting protein markers corresponding to different T cell activation statuses. Data generated by flow cytometry were corroborated by different antibody based assay technologies to detect soluble cytokines. Our findings suggest that there is an inter donor variability in both baseline and activation responses. We have also successfully developed an antibody panel for flow cytometry that could be used to study cytotoxic function of CD8 T cells in clinical immunology research areas.

Keywords

CD8+ cytotoxic PBMC Flow cytometry Antibody panel 

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Department of Medical Oncology, Center for Immuno-OncologyDana-Farber Cancer InstituteBostonUSA
  2. 2.Department of Medical Oncology/Hematologic NeoplasiaDana-Farber Cancer InstituteBostonUSA

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