Increased NK cell maturation in patients with acute myeloid leukemia

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Abstract

Understanding immune alterations in cancer patients is a major challenge and requires precise phenotypic study of immune subsets. Improvement of knowledge regarding the biology of natural killer (NK) cells and technical advances leads to the generation of high dimensional dataset. High dimensional flow cytometry requires tools adapted to complex dataset analyses. This study presents an example of NK cell maturation analysis in Healthy Volunteers (HV) and patients with Acute Myeloid Leukemia (AML) with an automated procedure using the FLOCK algorithm. This procedure enabled to automatically identify NK cell subsets according to maturation profiles, with 2D mapping of a four-dimensional dataset. Differences were highlighted in AML patients compared to HV, with an overall increase of NK maturation. Among patients, a strong heterogeneity in NK cell maturation defined three distinct profiles. Overall, automatic gating with FLOCK algorithm is a recent procedure, which enables fast and reliable identification of cell populations from high-dimensional cytometry data. Such tools are necessary for immune subset characterization and standardization of data analyses. This tool is adapted to new immune cell subsets discovery, and may lead to a better knowledge of NK cell defects in cancer patients. Overall, 2D mapping of NK maturation profiles enabled fast and reliable identification of NK cell subsets.

Figures

  • TABLE 1 | Patients characteristics.
  • FIGURE 1 | Comparison of manually gated data and FLOCK analysis. (A) NK clusters were automatically defined by FLOCK and manually annotated as CD56bright or CD56dim NK cells, among which four additional subsets were defined according to KIR and CD57 expression. For a given sample, clusters were defined by FLOCK. The clusters were merged when corresponding to the same NK subpopulation and visualized with FlowJo for comparison with manual gating. Each color represents the merging of clusters corresponding to the same population. (B) Frequencies of FLOCK and manually gated subsets of NK cells with respect to CD56, CD57, KIR expression. Data are presented as mean±SD of Healthy Volunteers (N= 18).
  • FIGURE 2 | NK maturation in HV and AML patients. NK maturation profiles in HV (A) and AML patients (B) were defined according to FLOCK output and NK subpopulations were represented using Manhattan Hierarchical Clustering based on CD56, KIR, NKG2A, and CD57 expression. Five clusters were defined; each individual is represented in three to five clusters, depending on the presence or absence of NK cells in the different clusters. Percentages of NK cells within clusters are presented as mean±SD in HV (C) and AML (D).
  • FIGURE 3 | AML patients can be classified into three distinct groups according to NK maturation profiles. Left panel: patients and HV were grouped according to the percentage of NK cells represented in the CD56bright, KIR−/CD57−, KIR+/CD57−, KIR−/CD57+, KIR+/CD57+ clusters using hierarchical

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Chretien, A. S., Granjeaud, S., Gondois-Rey, F., Harbi, S., Orlanducci, F., Blaise, D., … Olive, D. (2015). Increased NK cell maturation in patients with acute myeloid leukemia. Frontiers in Immunology, 6(NOV). https://doi.org/10.3389/fimmu.2015.00564

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