The NK Cell Curriculum: the education pathway to achieve polyfunctionality — ASN Events
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  3. The NK Cell Curriculum: the education pathway to achieve polyfunctionality

The NK Cell Curriculum: the education pathway to achieve polyfunctionality (#223)

Luis Romero Martin 1 , Valérie Monceaux 1 , Lechartier Marine 1 , Saez-Cirion Asier 1
  1. Institut Pasteur, Paris, ILE DE FRANCE, France

Background. Natural Killer (NK) cells are known for their cytotoxic potential and ability to produce various cytokines. NK cell responsiveness is influenced by inhibitory signals during education (via KIR, NKG2A, or both). However, the impact of NK cell education on polyfunctionality has been largely overlooked.

Methods. NK cells were isolated from heparinized blood from 15 donors of the EFS (Établissement Français du Sang) and stimulated for 4h in the presence of 721.221 cells as target. Single cell secretome was analyzed by the Isolight technology. The impact of NK cell education on polyfunctionality and phenotype was evaluated by a 40-color marker spectral flow cytometry panel.

Results. Principal component analysis (PCA) of the single cell secretome identified distinct clusters of NK cells based on their secretion of cytolytic molecules (Granzyme B and Perforin) or multiple cytokines. IL-17A and TGF-β were the most discriminative molecules. The expression of specific receptors had a divergent effect on the functional potential of educated CD56dimCD57− NK cells. KIR2DL1/KIR2DL2+ cells were associated with a more mature cytotoxic profile (Granzyme A, B, and M) but showed a weaker cytokine response. KIR3DL1+ cells were highly polyfunctional, producing 5/7 cytokines but had lower cytotoxic potential. KIR2DL5+ cells produced the highest levels of IL-10 and IL-17A, along with intermediate levels of granzymes. NKG2+ cells exhibited strong IFN-γ responses and high levels of perforin and Granzyme K, especially NKG2A+ cells. The co-expression of these molecules had an additive effect on their "predetermined" functional program, enabling cells to diversify their capacities. Differences related to expression of KIR and NKG2 receptors were less pronounced although still observable in differentiated CD57+ NK cells.

Conclusions. Our results indicate that early expression of different KIR or NKG2 receptors during development may restrict the cytolytic and cytokine program, resulting in the functional specialization of NK cells.

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  2. Lisovsky I, Kant S, Tremblay-McLean A, Isitman G, Kiani Z, Dupuy FP, Gilbert L, Bruneau J, Shoukry NH, Lebouché B, Bernard NF. Differential contribution of education through KIR2DL1, KIR2DL3, and KIR3DL1 to antibody-dependent (AD) NK cell activation and ADCC. J Leukoc Biol. 2019 Mar;105(3):551-563. doi: 10.1002/JLB.4A0617-242RRR. Epub 2019 Jan 30. PMID: 30698860; PMCID: PMC6916277.