title: Lysis-on-chip of single target cells following forced interaction with CTLs or NK cells on a dielectrophoresis-based array.
abstract: Guiding the interaction of single cells acting as partners in heterotypic interactions (e.g., effectors and targets of immune lysis) and monitoring the outcome of these interactions are regarded as crucial biomedical achievements. In this study, taking advantage of a dielectrophoresis (DEP)-based Laboratory-on-a-chip platform (the DEPArray), we show that it is possible to generate closed DEP cages entrapping CTLs and NK cells as either single cells or clusters; reversibly immobilize a single virus-presenting or tumor cell within the chip at a selected position; move cages and their content to predetermined spatial coordinates by software-guided routing; force a cytotoxic effector to physically interact with a putative target within a secluded area by merging their respective cages; generate cages containing effector and target cells at predetermined E:T ratios; accurately assess cytotoxicity by real-time quantitation of the release kinetics of the fluorescent dye calcein from target cells (>50 lytic events may be tested simultaneously); estimate end points of calcein release within 16 min of initial E:T cell contact; simultaneously deliver Ab-based phenotyping and on-chip lysis assessment; and identify lytic and nonlytic E:T combinations and discriminate nonlytic effector phenotypes from target refractoriness to immune lysis. The proof of principle is provided that DEPArray technology, previously used to levitate and move single cells, can be used to identify highly lytic antiviral CTLs and tumor cells that are particularly refractory to NK cell lysis. These findings are of primary interest in targeted immunotherapy.
authors.affiliation: Silicon Biosystems, 40129 Bologna, Italy;
authors.forename: Melanie
authors.initials: M
authors.lastname: Abonnenc
authors.forename: Monica
authors.initials: M
authors.lastname: Borgatti
authors.forename: Enrica
authors.initials: E
authors.lastname: Fabbri
authors.forename: Riccardo
authors.initials: R
authors.lastname: Gavioli
authors.forename: Cinzia
authors.initials: C
authors.lastname: Fortini
authors.forename: Federica
authors.initials: F
authors.lastname: Destro
authors.forename: Luigi
authors.initials: L
authors.lastname: Altomare
authors.forename: Nicolò
authors.initials: N
authors.lastname: Manaresi
authors.forename: Gianni
authors.initials: G
authors.lastname: Medoro
authors.forename: Aldo
authors.initials: A
authors.lastname: Romani
authors.forename: Marco
authors.initials: M
authors.lastname: Tartagni
authors.forename: Elisa
authors.initials: E
authors.lastname: Lo Monaco
authors.forename: Patrizio
authors.initials: P
authors.lastname: Giacomini
authors.forename: Roberto
authors.initials: R
authors.lastname: Guerrieri
authors.forename: Roberto
authors.initials: R
authors.lastname: Gambari
mesh_terms: D002450:Cell Communication / Q000276:immunology; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism