Designed DNA Surfaces for in Vitro Modulation of Natural Killer Cells

Abstract

Natural killer (NK) cells are at the junction of the innate and the adaptive immune response and play a very important role in host defense against viral infections and cancer. They have numerous cell surface receptors that activate or inhibit various intracellular signaling cascades that are then integrated to determine the functional activity of these cells. Here we present a surface-based approach that aims to tackle the largely unknown molecular mechanisms of signal integration. We use DNA microarrays containing capture oligonucleotides for the DNA-directed immobilization (DDI) of oligonucleotide-tagged αCD16 antibodies as ligands for NK cells. We demonstrate that the resulting surfaces can be gradually tuned in terms of ligand density to trigger the activation of living NK cells, as evidenced by degranulation, the release of cytokines, and intracellular Ca(2+) flux, measured at the level of single cells.

From DNA Nanotechnology to Material Systems Engineering

“…Furthermore, DNA‐directed patterning of single cells can be achieved by using the atomic force microscope for top‐down mechanical positioning of individual cells . The DNA‐based approaches have been exploited for electrochemical metabolic analysis of single mammalian cells on gold electrodes, the analysis of intercellular signaling, the multiplexed patterning of single cells for tissue engineering,[173a] the activation of natural killer cells, or for switch‐like, light‐controlled perturbations inside living cells …”

Biopebbles: DNA‐Functionalized Core–Shell Silica Nanospheres for Cellular Uptake and Cell Guidance Studies

“…Indeed, reversible formation of binary particle assemblies confirmed the integrity and functionality of the ssDNA‐conjugated particles (Figures S4 and S5, Supporting Information). To carry out hybridization experiments on solid surfaces, two orthogonal amino‐modified capture oligomers were spotted ( D3′ and D4′ ) as rectangular arrays (Figure S6, Supporting Information) on epoxy‐functionalized glass‐slides . Initial experiments were carried out with fluorescein‐containing SiNP 320 ‐7(D1/D3) , bearing two different oligonucleotides on their surface, which were allowed to hybridize and bind onto the ssDNA‐patterned substrates (Figure ).…”

DNA Surface Technology: From Gene Sensors to Integrated Systems for Life and Materials Sciences

“…In general, cell microarrays are important for tissue engineering, diagnostics, assays in drug development, and fundamental studies in cell biology . Indeed, specific case studies concerned the multiplexed patterning of single cells for tissue engineering, the metabolic analysis of single mammalian cells, the analysis of intercellular signaling, and the activation of natural killer cells …”