Automated cell enrichment of cytomegalovirus-specific T cells for clinical applications using the cytokine-capture system

Pappanaicken Kumaresan, Mathew Figliola, Judy S. Moyes, M. Helen Huls, Priti Tewari, Elizabeth J. Shpall, Richard Champlin, Laurence J N Cooper

Research output: Contribution to journalArticlepeer-review

20 Scopus citations


The adoptive transfer of pathogen-specific T cells can be used to prevent and treat opportunistic infections such as cytomegalovirus (CMV) infection occurring after allogeneic hematopoietic stem-cell transplantation. Viral-specific T cells from allogeneic donors, including third party donors, can be propagated ex vivo in compliance with current good manufacturing practice (cGMP), employing repeated rounds of antigendriven stimulation to selectively propagate desired T cells. The identification and isolation of antigen-specific T cells can also be undertaken based upon the cytokine capture system of T cells that have been activated to secrete gamma-interferon (IFN-γ). However, widespread human application of the cytokine capture system (CCS) to help restore immunity has been limited as the production process is time-consuming and requires a skilled operator. The development of a second-generation cell enrichment device such as CliniMACS Prodigy now enables investigators to generate viral-specific T cells using an automated, less labor-intensive system. This device separates magnetically labeled cells from unlabeled cells using magnetic activated cell sorting technology to generate clinical-grade products, is engineered as a closed system and can be accessed and operated on the benchtop. We demonstrate the operation of this new automated cell enrichment device to manufacture CMV pp65-specific T cells obtained from a steady-state apheresis product obtained from a CMV seropositive donor. These isolated T cells can then be directly infused into a patient under institutional and federal regulatory supervision. All the bio-processing steps including removal of red blood cells, stimulation of T cells, separation of antigen-specific T cells, purification, and washing are fully automated. Devices such as this raise the possibility that T cells for human application can be manufactured outside of dedicated good manufacturing practice (GMP) facilities and instead be produced in blood banking facilities where staff can supervise automated protocols to produce multiple products.

Original languageEnglish (US)
Article numbere52808
JournalJournal of Visualized Experiments
Issue number104
StatePublished - Oct 5 2015
Externally publishedYes


  • Anti-viral immunotherapy
  • Automated cell enrichment device
  • Bioprocessing
  • CMV-specific T cells
  • Cytokine-capture system (CCS)
  • IFN-gamma secreting T cells
  • Immunology
  • Issue 104
  • Magnetic-activated cell sorting technology
  • Pp65

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)


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