Frontiers in Immunology (Jul 2024)
A novel NKp80-based strategy for universal identification of normal, reactive and tumor/clonal natural killer-cells in blood
- F. Javier Morán-Plata,
- F. Javier Morán-Plata,
- Noemí Muñoz-García,
- Noemí Muñoz-García,
- María González-González,
- María González-González,
- Julio Pozo,
- Julio Pozo,
- Sonia Carretero-Domínguez,
- Sonia Carretero-Domínguez,
- Sonia Carretero-Domínguez,
- Sheila Mateos,
- Sheila Mateos,
- Susana Barrena,
- Susana Barrena,
- Moncef Belhassen-García,
- Moncef Belhassen-García,
- Catarina Lau,
- Maria Dos Anjos Teixeira,
- Ana Helena Santos,
- Ana Yeguas,
- Ana Balanzategui,
- Ana Balanzategui,
- Alejandro Martín García-Sancho,
- Alejandro Martín García-Sancho,
- Alejandro Martín García-Sancho,
- Alejandro Martín García-Sancho,
- Alberto Orfao,
- Alberto Orfao,
- Alberto Orfao,
- Alberto Orfao,
- Julia Almeida,
- Julia Almeida,
- Julia Almeida,
- Julia Almeida
Affiliations
- F. Javier Morán-Plata
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- F. Javier Morán-Plata
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Noemí Muñoz-García
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Noemí Muñoz-García
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- María González-González
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- María González-González
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Julio Pozo
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Julio Pozo
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Sonia Carretero-Domínguez
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Sonia Carretero-Domínguez
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Sonia Carretero-Domínguez
- Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain
- Sheila Mateos
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Sheila Mateos
- Cell-purification Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Susana Barrena
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Susana Barrena
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Moncef Belhassen-García
- Department of Internal Medicine, University Hospital of Salamanca, Salamanca, Spain
- Moncef Belhassen-García
- Department of Infectious Diseases, University Hospital of Salamanca, Centro de Investigación de Enfermedades Tropicales de la Universidad de Salamanca (CIETUS), Salamanca, Spain
- Catarina Lau
- Laboratory of Cytometry, Unit for Hematology Diagnosis, Department of Hematology, Hospital de Santo António (HSA), Centro Hospitalar Universitário do Porto (CHUP), Unidade Multidisciplinar de Investigação Biomédica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (UMIB/ICBAS/UP), Porto, Portugal
- Maria Dos Anjos Teixeira
- Laboratory of Cytometry, Unit for Hematology Diagnosis, Department of Hematology, Hospital de Santo António (HSA), Centro Hospitalar Universitário do Porto (CHUP), Unidade Multidisciplinar de Investigação Biomédica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (UMIB/ICBAS/UP), Porto, Portugal
- Ana Helena Santos
- Laboratory of Cytometry, Unit for Hematology Diagnosis, Department of Hematology, Hospital de Santo António (HSA), Centro Hospitalar Universitário do Porto (CHUP), Unidade Multidisciplinar de Investigação Biomédica, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (UMIB/ICBAS/UP), Porto, Portugal
- Ana Yeguas
- Department of Hematology, University Hospital of Salamanca, Salamanca, Spain
- Ana Balanzategui
- Department of Hematology, University Hospital of Salamanca, Salamanca, Spain
- Ana Balanzategui
- Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain
- Alejandro Martín García-Sancho
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Alejandro Martín García-Sancho
- Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain
- Alejandro Martín García-Sancho
- Department of Hematology, University Hospital of Salamanca, Salamanca, Spain
- Alejandro Martín García-Sancho
- Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain
- Alberto Orfao
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Alberto Orfao
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Alberto Orfao
- Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain
- Alberto Orfao
- Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain
- Julia Almeida
- Translational and Clinical Research Program, Cancer Research Center (IBMCC, CSIC – University of Salamanca), and Department of Medicine, University of Salamanca, Salamanca, Spain
- Julia Almeida
- Cytometry Service, NUCLEUS, University of Salamanca, Salamanca, Spain
- Julia Almeida
- Institute of Biomedical Research of Salamanca (IBSAL), Salamanca, Spain
- Julia Almeida
- Research Networking Centre Consortium of Oncology (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain
- DOI
- https://doi.org/10.3389/fimmu.2024.1423689
- Journal volume & issue
-
Vol. 15
Abstract
PurposeNatural killer (NK) cells are traditionally identified by flow cytometry using a combination of markers (CD16/CD56/CD3), because a specific NK-cell marker is still missing. Here we investigated the utility of CD314, CD335 and NKp80, compared to CD16/CD56/CD3, for more robust identification of NK-cells in human blood, for diagnostic purposes.MethodsA total of 156 peripheral blood (PB) samples collected from healthy donors (HD) and patients with diseases frequently associated with loss/downregulation of classical NK-cell markers were immunophenotyped following EuroFlow protocols, aimed at comparing the staining profile of total blood NK-cells for CD314, CD335 and NKp80, and the performance of distinct marker combinations for their accurate identification.ResultsNKp80 showed a superior performance (vs. CD314 and CD335) for the identification of NK-cells in HD blood. Besides, NKp80 improved the conventional CD16/CD56/CD3-based strategy to identify PB NK-cells in HD and reactive processes, particularly when combined with CD16 for further accurate NK-cell-subsetting. Although NKp80+CD16 improved the identification of clonal/tumor NK-cells, particularly among CD56- cases (53%), aberrant downregulation of NKp80 was observed in 25% of patients, in whom CD56 was useful as a complementary NK-cell marker. As NKp80 is also expressed on T-cells, we noted increased numbers of NKp80+ cytotoxic T-cells at the more advanced maturation stages, mostly in adults.ConclusionHere we propose a new robust approach for the identification of PB NK-cells, based on the combination of NKp80 plus CD16. However, in chronic lymphoproliferative disorders of NK-cells, addition of CD56 is recommended to identify clonal NK-cells, due to their frequent aberrant NKp80- phenotype.
Keywords
