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Quantitative proteomics of cytokine-induced memory-like (CIML) NK cells

ABSTRACT: NK cells from healthy donors were isolated from peripheral blood and stimulated overnight with IL-12/15/18 to induce a memory-like phenotype. After 7 days of culture, the phenotype and degranulation potential of CIML NK cells was characterized in two distinct NK cell subsets: CD16+/CD56+ and CD16−/CD56. Finally, NK cell subsets were purified using fluorescence-activated cell sorting (FACS) and subjected to high-throughput multiplexed quantitative proteomics.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Natural Killer Cell Activation Involved In Immune Response

SUBMITTER: Inmaculada Jorge

LAB HEAD: Jesús María Vázquez Cobos

PROVIDER: PXD059190 | Pride | 2025-07-14

REPOSITORIES: Pride

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Project description:Human Natural Killer (NK) cells comprise two main subsets, CD56bright and CD56dim cells, that differ in function, phenotype and tissue localization. To further dissect the heterogeneity of CD56dim cells, we have performed transcriptome analysis and functional ex vivo characterization of human NK cell subsets according to the expression of markers related to differentiation, migration or competence. Here, we show for the first time that the ability to respond to cytokines or to activating receptors is mutually exclusive in almost all NK cells with the exception of CD56dim CD62L+ cells. Indeed, only these cells combine the ability to produce interferon (IFN)-gamma after cytokines and proliferate in vivo during viral infection with the capacity to kill and produce cytokines upon engagement of activating receptors. Therefore, CD56dim CD62L+ cells represent a unique subset of polyfunctional NK cells. Ex vivo analysis of their function, phenotype, telomere length, frequencies during ageing as well as transfer experiments of NK cell subsets into immunodeficient mice suggest that CD56dim CD62L+ cells represent an intermediate stage of NK cell maturation, which after restimulation can accomplish multiple tasks and further develop into terminally differentiated effectors. Gene expression profiles of FACSAria sorted CD3- CD56bright CD62L+, CD3- CD56dim CD62L+ and CD3- CD56dim CD62L- NK cells from human peripheral blood of three donors were compared using Affymetrix GeneChip Human Genome HG-U133_Plus_2. After total RNA extraction, reverse transcription, cDNA extraction, the biotinylated cRNA was transcribed, fragmented, and 15 Âµg cRNA hybridized in triplicates for each of the three groups to the GeneChip arrays. Group1: CD3- CD56bright CD62L+,.Group2: CD3- CD56dim CD62L+, Group3: CD3- CD56dim CD62L-. Lists of differentially regulated genes were created using High Performance Chip Data Analysis (HPCDA) with Bioretis database (http://www.bioretis-analysis.de). Worldwide data sharing is possible via Bioretis, please ask the authors.

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	Aliq_1_IJ.msf	Msf
	Aliq_1_IJ.raw	Raw
	Aliq_2_IJ.msf	Msf
	Aliq_2_IJ.raw	Raw
	FR1_1_IJ.msf	Msf

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Quantitative proteomics of cytokine-induced memory-like (CIML) NK cells

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