Quantitative analysis of human NK cell reactivity using latex beads coated with defined amounts of antibodies.,European Journal of Immunology

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Quantitative analysis of human NK cell reactivity using latex beads coated with defined amounts of antibodies.
European Journal of Immunology ( IF 4.5 ) Pub Date : 2020-02-19 , DOI: 10.1002/eji.201948344
Madeleine Dorsch ₁ , Doris Urlaub ₁ , Vivian Bönnemann ₁ , Peter Bröde ₁ , Mina Sandusky ₁ , Carsten Watzl ₁

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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.

中文翻译：

使用涂有确定量抗体的乳胶珠对人类NK细胞反应性进行定量分析。

天然杀伤（NK）细胞反应受多种不同的表面受体调节。尽管我们可以确定给定受体对NK细胞功能的总体正面或负面影响，但以定量方式研究NK细胞的调控却具有挑战性。为了定量研究各个受体对NK细胞活化的影响，我们选择功能化的乳胶珠，其大小与淋巴细胞大致相同，并具有确定量的针对不同活化受体的特异性抗体。这使我们能够在定义，清洁和可控制的系统中研究NK细胞的反应性。只有CD16和NKp30可以激活静止的人NK细胞的脱颗粒。CD16，NKG2D，NKp30，NKp44和NKp46能够激活培养的NK细胞。NK细胞激活导致脱粒并产生IFN-γ和MIP-1β的多功能细胞的诱导。有趣的是，多功能NK细胞仅通过触发ITAM偶联受体来诱导。NKp44显示非常敏感的响应模式，其中受体刺激的少量增加导致最大的NK细胞活性。相反，刺激2B4诱导NK细胞脱粒非常少，同时为NK细胞粘附提供足够的信号。我们的数据表明，激活受体在刺激NK细胞方面的功效不同。相反，刺激2B4诱导NK细胞脱粒非常少，同时为NK细胞粘附提供足够的信号。我们的数据表明，激活受体在刺激NK细胞方面的功效不同。相反，刺激2B4诱导NK细胞脱粒非常少，同时为NK细胞粘附提供足够的信号。我们的数据表明，激活受体在刺激NK细胞方面的功效不同。

更新日期：2020-02-19

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