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Quantitative analysis of human NK cell reactivity using latex beads coated with defined amounts of antibodies
Author(s) -
Dorsch Madeleine,
Urlaub Doris,
Bönnemann Vivian,
Bröde Peter,
Sandusky Mina,
Watzl Carsten
Publication year - 2020
Publication title -
european journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.272
H-Index - 201
eISSN - 1521-4141
pISSN - 0014-2980
DOI - 10.1002/eji.201948344
Subject(s) - degranulation , nkg2d , cd16 , receptor , microbiology and biotechnology , biology , cell , lymphokine activated killer cell , natural killer cell , stimulation , antibody , interleukin 21 , interleukin 12 , immunology , in vitro , cytotoxicity , immune system , t cell , cytotoxic t cell , biochemistry , cd8 , endocrinology , cd3
Abstract Natural Killer (NK) cell responses are regulated by a variety of different surface receptors. While we can determine the overall positive or negative effect of a given receptor on NK cell functions, investigating NK cell regulation in a quantitative way is challenging. To quantitatively investigate individual receptors for their effect on NK cell activation, we chose to functionalize latex beads that have approximately the same size as lymphocytes with defined amounts of specific antibodies directed against distinct activating receptors. This enabled us to investigate NK cell reactivity in a defined, clean, and controllable system. Only CD16 and NKp30 could activate the degranulation of resting human NK cells. CD16, NKG2D, NKp30, NKp44, and NKp46 were able to activate cultured NK cells. NK cell activation resulted in the induction of polyfunctional cells that degranulated and produced IFN‐γ and MIP‐1β. Interestingly, polyfunctional NK cells were only induced by triggering ITAM‐coupled receptors. NKp44 showed a very sensitive response pattern, where a small increase in receptor stimulation caused maximal NK cell activity. In contrast, stimulation of 2B4 induced very little NK cell degranulation, while providing sufficient signal for NK cell adhesion. Our data demonstrate that activating receptors differ in their effectiveness to stimulate NK cells.