Isolation of tumour-reactive lymphocytes from peripheral blood via microfluidic immunomagnetic cell sorting

Zongjie Wang, Sharif Ahmed, Mahmoud Aziz Mahmoud Labib, Hansen Wang, Licun Wu, Fatemeh Bavaghar-Zaeimi, Nastaran Shokri, Soraly Blanco, Saraf Karim, Kasia Czarnecka-Kujawa, Edward H. Sargent, A. J.Robert McGray, Marc de Perrot, Shana O. Kelley*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

The clinical use of tumour-infiltrating lymphocytes for the treatment of solid tumours is hindered by the need to obtain large and fresh tumour fractions, which is often not feasible in patients with unresectable tumours or recurrent metastases. Here we show that circulating tumour-reactive lymphocytes (cTRLs) can be isolated from peripheral blood at high yield and purity via microfluidic immunomagnetic cell sorting, allowing for comprehensive downstream analyses of these rare cells. We observed that CD103 is strongly expressed by the isolated cTRLs, and that in mice with subcutaneous tumours, tumour-infiltrating lymphocytes isolated from the tumours and rapidly expanded CD8+CD103+ cTRLs isolated from blood are comparably potent and respond similarly to immune checkpoint blockade. We also show that CD8+CD103+ cTRLs isolated from the peripheral blood of patients and co-cultured with tumour cells dissociated from their resected tumours resulted in the enrichment of interferon-γ-secreting cell populations with T-cell-receptor clonotypes substantially overlapping those of the patients’ tumour-infiltrating lymphocytes. Therapeutically potent cTRLs isolated from peripheral blood may advance the clinical development of adoptive cell therapies.

Original languageEnglish (US)
Pages (from-to)1188-1203
Number of pages16
JournalNature Biomedical Engineering
Volume7
Issue number9
DOIs
StatePublished - Sep 2023

Funding

We thank T. Chen at the Centre for Advanced Single Cell Analysis (CASCA), Sick Children Hospital, Toronto for her help in CyTOF, N. Simard at the centralize flow cytometry facility at Temerty Faculty of Medicine, University of Toronto for her help in FACS sorting, A. C. Zhou at the Medicine by Design initiative at the University of Toronto for her comments, W. Xiao and A. Archila at the University Health Network (UHN) for their help in tail vein injection, M. Peralta at the UHN PRP facility and N. Law at the UHN STTARR facility for their help in IHC, J. Jonkman at the UHN AOMF facility for his help in image quantitation, and J. Wei and J. Moffat at the Terrence Donnelly Centre, University of Toronto for donating CT26HAand OT-1 cells. This study was supported in part by the Canadian Institutes of Health Research (grant no. FDN-148415) and the Collaborative Health Research Projects program (CIHR/NSERC partnered). This research was part of the University of Toronto’s Medicine by Design initiative, which receives funding from the Canada First Research Excellence Fund. The study was also supported in part by the McCormick Catalyst Fund at Northwestern University. We thank T. Chen at the Centre for Advanced Single Cell Analysis (CASCA), Sick Children Hospital, Toronto for her help in CyTOF, N. Simard at the centralize flow cytometry facility at Temerty Faculty of Medicine, University of Toronto for her help in FACS sorting, A. C. Zhou at the Medicine by Design initiative at the University of Toronto for her comments, W. Xiao and A. Archila at the University Health Network (UHN) for their help in tail vein injection, M. Peralta at the UHN PRP facility and N. Law at the UHN STTARR facility for their help in IHC, J. Jonkman at the UHN AOMF facility for his help in image quantitation, and J. Wei and J. Moffat at the Terrence Donnelly Centre, University of Toronto for donating CT26 and OT-1 cells. This study was supported in part by the Canadian Institutes of Health Research (grant no. FDN-148415) and the Collaborative Health Research Projects program (CIHR/NSERC partnered). This research was part of the University of Toronto’s Medicine by Design initiative, which receives funding from the Canada First Research Excellence Fund. The study was also supported in part by the McCormick Catalyst Fund at Northwestern University. HA S.O.K. and Z.W. have a filled patent application using parts of the data reported in this article. S.O.K. has a patent ‘Device for capture of particles in a flow’ US10073079 licensed to Cellular Analytics. A.J.R.M. is a paid consultant for Cellular Analytics. M.D.P. received personal fees from Actelion, AstraZeneca, Bayer, Bristol Myers Squibb, Merck and Roche outside of the submitted work. S.O.K. received research funds from Amgen through a sponsored research agreement. The other authors declare no competing interests.

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering
  • Computer Science Applications

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