title: Microfluidic devices modulate tumor cell line susceptibility to NK cell recognition.
abstract: This study aims to adoptively reduce the major histocompatibility complex class I (MHC-I) molecule surface expression of cancer cells by exposure to microfluid shear stress and a monoclonal antibody. A microfluidic system is developed and tumor cells are injected at different flow rates. The bottom surface of the microfluidic system is biofunctionalized with antibodies (W6/32) specific for the MHC-I molecules with a simple method based on microfluidic protocols. The antibodies promote binding between the bottom surface and the MHC-I molecules on the tumor cell membrane. The cells are injected at an optimized flow rate, then roll on the bottom surface and are subjected to shear stress. The stress is localized and enhanced on the part of the membrane where MHC-I proteins are expressed, since they stick to the antibodies of the system. The localized stress allows a stripping effect and consequent reduction of the MHC-I expression. It is shown that it is possible to specifically treat and recover eukaryotic cells without damaging the biological samples. MHC-I molecule expression on treated and control cell surfaces is measured on tumor and healthy cells. After the cell rolling treatment a clear reduction of MHC-I levels on the tumor cell membrane is observed, whereas no changes are observed on healthy cells (monocytes). The MHC-I reduction is investigated and the possibility that the developed system could induce a loss of these molecules from the tumor cell surface is addressed. The percentage of living tumor cells (viability) that remain after the treatment is measured. The changes induced by the microfluidic system are analyzed by fluorescence-activated cell sorting and confocal microscopy. Cytotoxicity tests show a relevant increased susceptibility of natural killer (NK) cells on microchip-treated tumor cells.
authors: BioNEM (Bio and Nano Engineering and Technology for Medicine) Laboratory, Department of Clinical and Experimental Medicine, University "Magna Graecia" of Catanzaro, Loc. Germaneto, 88100 Catanzaro, Italy. Gerardo.Perozziello@unicz.it
authors: Gerardo
authors: G
authors: Perozziello
authors: Rosanna
authors: R
authors: La Rocca
authors: Gheorghe
authors: G
authors: Cojoc
authors: Carlo
authors: C
authors: Liberale
authors: Natalia
authors: N
authors: Malara
authors: Giuseppina
authors: G
authors: Simone
authors: Patrizio
authors: P
authors: Candeloro
authors: Andrea
authors: A
authors: Anichini
authors: Luca
authors: L
authors: Tirinato
authors: Francesco
authors: F
authors: Gentile
authors: Maria Laura
authors: ML
authors: Coluccio
authors: Ennio
authors: E
authors: Carbone
authors: Enzo
authors: E
authors: Di Fabrizio
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011485:Protein Binding
chemical_list: D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I