Measuring Chimeric Antigen Receptor T Cells (CAR T Cells) Activation by Coupling Intracellular Cytokine Staining with Flow Cytometry

  • Chong XuEmail author
  • Yibo YinEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2108)


Chimeric antigen receptor T cells (CAR T cells) therapy is one kind of immunotherapy that has revolutionally changed the landscape of immunotherapy and been approved by the FDA from 2017 for several blood malignancies. To bring new CAR T cells to clinic, every new CAR need to test in vitro for antigen recognition, tumor cell killing capacity, and off-target cytotoxicity effect. Detecting the secretion of cytokines upon engagement of CAR T cells with tumor antigens is routinely applied to assess these CAR functions. Here we describe coupling of intracellular cytokine staining and multicolor flow cytometry to measure CAR T cells activation upon antigen stimulation.

Key words

Chimeric antigen receptor T cells CAR T cells Intracellular cytokine staining Flow cytometry T cell activation Immunotherapy 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Center for Cellular Immunotherapies, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.The Parker Institute for Cancer Immunotherapy, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  3. 3.Department of Pathology and Laboratory Medicine, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  4. 4.Department of Medicine, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  5. 5.Department of Neurosurgery, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  6. 6.The Fourth Section of Department of NeurosurgeryThe First Affiliated Hospital of Harbin Medical UniversityHarbinP.R. China

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