Single-cell cloning of NK cells from rhesus macaques: a new methodology for in-depth analysis of MHC-E-dependent adaptive functions

Abstract Natural killer (NK) cells provide rapid and robust responses against a wide range of viral infections. More recently, multiple studies have shown that subsets NK are also capable antigen-specific recall to eliminate infected cells. In humans nonhuman primates (NHP), memory cell can at least be partially delineated by expression NKG2C, ligand for MHC-E. Due MHC-E limited polymorphism, MHC-E-presented antigens may induce universally To study in NHP, we developed new vitromethod clonally expand single rhesus macaque (rhNKCL) from blood spleen. define the effector functions rhNKCL, used calcein acetoxymethyl ester (CAM) cytotoxicity assay intracellular staining combination with K562 lines or autologous BLCL as targets. We identified, silicomethodology, peptides influenza, SIV, SARS-CoV-2 viruses predicted display strong binding Our procedure showed first-time clonal expansion observed conserved NHP phenotype CD3 −NKG2A/C +CD16 +CD56 +following additional surface receptors CD8, NKp80, NKG2D. Upon stimulation cells, rhNKCL (cytotoxicity, CD107a, IFNγ, TNFα) were like primary Altogether, this method single-cell cloning macaques could key information about including epigenetic, metabolic, transcriptional, phenotypic profiles, which exploited further enhance antiviral activity future vaccine strategies therapies. Supported grants NIH (R01 AI158516, R01 DK130472, AI161010, NIH-funded BEAT-HIV Martin Delaney Collaboratory cure HIV-1 infection Combination Immunotherapy UM1 AI164570).