title: Microchip-Based Single-Cell Imaging Reveals That CD56dimCD57-KIR-NKG2A+ NK Cells Have More Dynamic Migration Associated with Increased Target Cell Conjugation and Probability of Killing Compared to CD56dimCD57-KIR-NKG2A- NK Cells.
abstract: NK cells are functionally educated by self-MHC specific receptors, including the inhibitory killer cell Ig-like receptors (KIRs) and the lectin-like CD94/NKG2A heterodimer. Little is known about how NK cell education influences qualitative aspects of cytotoxicity such as migration behavior and efficacy of activation and killing at the single-cell level. In this study, we have compared the behavior of FACS-sorted CD56(dim)CD57(-)KIR(-)NKG2A(+) (NKG2A(+)) and CD56(dim)CD57(-)KIR(-)NKG2A(-) (lacking inhibitory receptors; IR(-)) human NK cells by quantifying migration, cytotoxicity, and contact dynamics using microchip-based live cell imaging. NKG2A(+) NK cells displayed a more dynamic migration behavior and made more contacts with target cells than IR(-) NK cells. NKG2A(+) NK cells also more frequently killed the target cells once a conjugate had been formed. NK cells with serial killing capacity were primarily found among NKG2A(+) NK cells. Conjugates involving IR(-) NK cells were generally more short-lived and IR(-) NK cells did not become activated to the same extent as NKG2A(+) NK cells when in contact with target cells, as evident by their reduced spreading response. In contrast, NKG2A(+) and IR(-) NK cells showed similar dynamics in terms of duration of conjugation periods and NK cell spreading response in conjugates that led to killing. Taken together, these observations suggest that the high killing capacity of NKG2A(+) NK cells is linked to processes regulating events in the recognition phase of NK-target cell contact rather than events after cytotoxicity has been triggered.
authors: Department of Microbiology, Tumor and Cell Biology, Science for Life Laboratory, Karolinska Institutet, 171 65 Stockholm, Sweden;
authors: Elin
authors: E
authors: Forslund
authors: Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 141 86 Stockholm, Sweden;
authors: Ebba
authors: E
authors: Sohlberg
authors: Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 141 86 Stockholm, Sweden;
authors: Monika
authors: M
authors: Enqvist
authors: Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, 171 65 Stockholm, Sweden;
authors: Per E
authors: PE
authors: Olofsson
authors: Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, 141 86 Stockholm, Sweden; Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital, Montebello, 0310 Oslo, Norway; and K.G. Jebsen Center for Cancer Immunotherapy, Institute of Clinical Medicine, University of Oslo, Montebello, 0310 Oslo, Norway.
authors: Karl-Johan
authors: KJ
authors: Malmberg
authors: Department of Microbiology, Tumor and Cell Biology, Science for Life Laboratory, Karolinska Institutet, 171 65 Stockholm, Sweden; Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, 171 65 Stockholm, Sweden; bjorn.onfelt@ki.se.
authors: Björn
authors: B
authors: Önfelt
mesh_terms: D019002:CD56 Antigen / Q000378:metabolism; D018959:CD57 Antigens / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D005434:Flow Cytometry; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007694:Killer Cells, Natural / Q000276:immunology*; D046208:Microchip Analytical Procedures; D055654:NK Cell Lectin-Like Receptor Subfamily C / Q000096:biosynthesis / Q000276:immunology*; D054340:Receptors, KIR / Q000378:metabolism
chemical_list: D019002:CD56 Antigen; D018959:CD57 Antigens; C527054:KLRC1 protein, human; C522918:NCAM1 protein, human; D055654:NK Cell Lectin-Like Receptor Subfamily C; D054340:Receptors, KIR