TY - JOUR
T1 - Isolation of tumour-reactive lymphocytes from peripheral blood via microfluidic immunomagnetic cell sorting
AU - Wang, Zongjie
AU - Ahmed, Sharif
AU - Labib, Mahmoud Aziz Mahmoud
AU - Wang, Hansen
AU - Wu, Licun
AU - Bavaghar-Zaeimi, Fatemeh
AU - Shokri, Nastaran
AU - Blanco, Soraly
AU - Karim, Saraf
AU - Czarnecka-Kujawa, Kasia
AU - Sargent, Edward H.
AU - McGray, A. J.Robert
AU - de Perrot, Marc
AU - Kelley, Shana O.
N1 - Funding Information:
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.
Funding Information:
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
Funding Information:
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.
Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2023/9
Y1 - 2023/9
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=85152411256&partnerID=8YFLogxK
U2 - 10.1038/s41551-023-01023-3
DO - 10.1038/s41551-023-01023-3
M3 - Article
C2 - 37037966
AN - SCOPUS:85152411256
SN - 2157-846X
VL - 7
SP - 1188
EP - 1203
JO - Nature Biomedical Engineering
JF - Nature Biomedical Engineering
IS - 9
ER -