Identification, sorting and profiling of functional killer cells via the capture of fluorescent target-cell lysate.

Journal

Nature biomedical engineering
ISSN: 2157-846X
Titre abrégé: Nat Biomed Eng
Pays: England
ID NLM: 101696896

Informations de publication

Date de publication:
31 Aug 2023
Historique:
received: 15 07 2022
accepted: 04 08 2023
medline: 1 9 2023
pubmed: 1 9 2023
entrez: 31 8 2023
Statut: aheadofprint

Résumé

Assays for assessing cell-mediated cytotoxicity are largely target-cell-centric and cannot identify and isolate subpopulations of cytotoxic effector cells. Here we describe an assay compatible with flow cytometry for the accurate identification and sorting of functional killer-cell subpopulations in co-cultures. The assay, which we named PAINTKiller (for 'proximity affinity intracellular transfer identification of killer cells'), relies on the detection of an intracellular fluorescent protein 'painted' by a lysed cell on the surface of the lysing cytotoxic cell (specifically, on cell lysis the intracellular fluorescein derivative carboxyfluorescein succinimidyl ester is captured on the surface of the natural killer cell by an antibody for anti-fluorescein isothiocyanate linked to an antibody for the pan-leucocyte surface receptor CD45). The assay can be integrated with single-cell RNA sequencing for the analysis of molecular pathways associated with cell cytotoxicity and may be used to uncover correlates of functional immune responses.

Identifiants

DOI: 10.1038/s41551-023-01089-z PMID: 37652987
pubmed: 37652987
doi: 10.1038/s41551-023-01089-z
pii: 10.1038/s41551-023-01089-z
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : MOH | National Medical Research Council (NMRC)
ID : MOH-OFIRG18nov-003
Organisme : Ministry of Education - Singapore (MOE)
ID : MOE-T2EP30120-0008

Informations de copyright

© 2023. The Author(s), under exclusive licence to Springer Nature Limited.

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Auteurs

Yen Hoon Luah (YH)

Institute for Health Innovation and Technology, National University of Singapore, Singapore, Singapore.
Critical Analytics for Manufacturing of Personalized-Medicine Interdisciplinary Research Group, Singapore-MIT Alliance in Research and Technology, Singapore, Singapore.

Tongjin Wu (T)

Institute for Health Innovation and Technology, National University of Singapore, Singapore, Singapore.
Department of Biomedical Engineering, College of Design and Engineering, National University of Singapore, Singapore, Singapore.
ORCID: 0000-0002-1233-4994

Lih Feng Cheow (LF)

Institute for Health Innovation and Technology, National University of Singapore, Singapore, Singapore. lihfeng.cheow@nus.edu.sg.
Critical Analytics for Manufacturing of Personalized-Medicine Interdisciplinary Research Group, Singapore-MIT Alliance in Research and Technology, Singapore, Singapore. lihfeng.cheow@nus.edu.sg.
Department of Biomedical Engineering, College of Design and Engineering, National University of Singapore, Singapore, Singapore. lihfeng.cheow@nus.edu.sg.
ORCID: 0000-0003-1766-0024

Classifications MeSH