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Author Notes:

To whom correspondence should be addressed: Gabe Kwong, Address: 345 Ferst Dr, Atlanta, GA 30332, Phone: 404-385-3746, gkwong@gatech.edu, ORCID:0000-0002-6255-6755.

We thank D. Arafat for assistance with ddPCR.

Subjects:

Research Funding:

This work was funded by an NIH Director’s New Innovator Award (Award No. DP2HD091793) and by the National Center for Advancing Translational Sciences of the NIH (Award No. UL1TR000454).

S.N.D. is supported by the NSF Graduate Research Fellowships Program (Grant No. DGE-1650044) and the NSF Integrative Graduate Education and Research Traineeship (Grant No. DGE-0965945).

Y.M.C. is supported by the National Institute of General Medical Sciences of the NIH (Award No. T32GM8169–30).

G.A.K. holds a Career Award at the Scientific Interface from the Burroughs Wellcome Fund.

This work was performed in part at the Georgia Tech Institute for Electronics and Nanotechnology, a member of the National Nanotechnology Coordinated Infrastructure, which is supported by the NSF (Grant No. ECCS-1542174).

Keywords:

  • Science & Technology
  • Physical Sciences
  • Chemistry, Analytical
  • Chemistry
  • TUMOR-CELLS
  • DYNAMICS
  • BLOOD
  • ACTIVATION
  • RECEPTORS
  • RESPONSES
  • PLATFORM

DNA-Barcoded pMHC Tetramers for Detection of Single Antigen-Specific T Cells by Digital PCR

Tools:

Journal Title:

Analytical Chemistry

Volume:

Volume 91, Number 4

Publisher:

, Pages 2695-2700

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Antigen-specific T cells are found at low frequencies in circulation but carry important diagnostic information as liquid biomarkers in numerous biomedical settings, such as monitoring the efficacy of vaccines and cancer immunotherapies. To enable detection of antigen-specific T cells with high sensitivity, we develop peptide-MHC (pMHC) tetramers labeled with DNA barcodes to detect single T cells by droplet digital PCR (ddPCR). We show that site-specific conjugation of DNA via photocleavable linkers allows barcoded tetramers to stain T cells with similar avidity compared to conventional fluorescent tetramers and efficient recovery of barcodes by light with no loss in cell viability. We design an orthogonal panel of DNA-barcoded tetramers to simultaneously detect multiple antigen-specific T cell populations, including from a mouse model of viral infection, and discriminate single cancer-specific T cells with high diagnostic sensitivity and specificity. This approach of DNA-barcoding can be broadened to encompass additional rare cells for monitoring immunological health at the single cell level.

Copyright information:

© 2019 American Chemical Society.

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