4.8 Article

Rapid, scalable assessment of SARS-CoV-2 cellular immunity by whole-blood PCR

期刊

NATURE BIOTECHNOLOGY
卷 40, 期 11, 页码 1680-+

出版社

NATURE PORTFOLIO
DOI: 10.1038/s41587-022-01347-6

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资金

  1. ISMMS seed fund
  2. Dean's office grant
  3. National Cancer Institute (NCI) Cancer Center Support grant [P30 CA196521]
  4. NCI [T32CA078207]
  5. Charles H. Revson Foundation
  6. Instituto de Salud Carlos III [COV20-00668, PI16CIII/00012]
  7. Instituto de Salud Carlos III, Spanish Ministry of Science and Innovation [COV20/00181]
  8. European Development Regional Fund 'A way to achieve Europe'
  9. Instituto de Salud Carlos III, Spain [COV20/00170]
  10. Government of Cantabria, Spain [2020UIC22-PUB-0019]
  11. Fondo Social Europeo e Iniciativa de Empleo Juvenil YEI [PEJ2018-004557-A]
  12. European Union [101037867]
  13. [U24CA224319]
  14. [U01DK124165]
  15. [NCI K00CA212474]
  16. [REDInREN 016/009/009 ISCIII]

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This study reports the development of two quantitative PCR assays for detecting SARS-CoV-2-specific T cell activation. These assays are rapid, internally normalized, and can quantify the magnitude and duration of cellular immune responses, helping to determine revaccination strategies in vulnerable populations.
The T cell response to SARS-CoV-2 is detected by a PCR assay on whole blood. Fast, high-throughput methods for measuring the level and duration of protective immune responses to SARS-CoV-2 are needed to anticipate the risk of breakthrough infections. Here we report the development of two quantitative PCR assays for SARS-CoV-2-specific T cell activation. The assays are rapid, internally normalized and probe-based: qTACT requires RNA extraction and dqTACT avoids sample preparation steps. Both assays rely on the quantification of CXCL10 messenger RNA, a chemokine whose expression is strongly correlated with activation of antigen-specific T cells. On restimulation of whole-blood cells with SARS-CoV-2 viral antigens, viral-specific T cells secrete IFN-gamma, which stimulates monocytes to produce CXCL10. CXCL10 mRNA can thus serve as a proxy to quantify cellular immunity. Our assays may allow large-scale monitoring of the magnitude and duration of functional T cell immunity to SARS-CoV-2, thus helping to prioritize revaccination strategies in vulnerable populations.

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