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( A ) The IFN-⍺ + , TNFα + , and GrzB + population (%) of <t>IL-2–activated</t> NK cells after antibodies treatment with different NKG2A binding valency (monovalent, bivalent, tetravalent, or multivalent) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Internalization of NKG2A antibodies in NK cells, assessed by labeling the antibody with a pH-sensitive dye to monitor endocytosis. Durvalumab and pertuzumab (ptz IgG1) were included as controls. (A and B) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.
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( A ) The IFN-⍺ + , TNFα + , and GrzB + population (%) of IL-2–activated NK cells after antibodies treatment with different NKG2A binding valency (monovalent, bivalent, tetravalent, or multivalent) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Internalization of NKG2A antibodies in NK cells, assessed by labeling the antibody with a pH-sensitive dye to monitor endocytosis. Durvalumab and pertuzumab (ptz IgG1) were included as controls. (A and B) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Journal: Science Advances

Article Title: Discovery and preclinical evaluation of monoclonal antibodies and bispecific engagers targeting the NKG2A inhibitory receptor

doi: 10.1126/sciadv.adu0690

Figure Lengend Snippet: ( A ) The IFN-⍺ + , TNFα + , and GrzB + population (%) of IL-2–activated NK cells after antibodies treatment with different NKG2A binding valency (monovalent, bivalent, tetravalent, or multivalent) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Internalization of NKG2A antibodies in NK cells, assessed by labeling the antibody with a pH-sensitive dye to monitor endocytosis. Durvalumab and pertuzumab (ptz IgG1) were included as controls. (A and B) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Primary NK cells were cultured with magnetic-activated cell sorting (MACS) basal NK media with supplementary (Miltenyi Biotec, 130-114-429), 10% (v/v) human serum (Sigma-Aldrich), and human IL-2 (hIL-2) (50 IU/ml; Miltenyi Biotec).

Techniques: Binding Assay, Labeling

( A ) CD16 + , NKG2A + , or NKG2C + population (%) of CD56 + NK cells after antibodies (100 nM) and IL-2 treatment (50 IU/ml) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Left: Graphs showing the cell killing activity of aHER2 × aNKG2A BiNK cotreatment with anti-HER2 IgG1 pertuzumab (ptz IgG1) in A549 and H2030 cells in the presence of primary NK cells. E:T ratio of 2:1. Right: Schematic figure of antibodies treatment schedule. (A) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05 and ** P < 0.01.

Journal: Science Advances

Article Title: Discovery and preclinical evaluation of monoclonal antibodies and bispecific engagers targeting the NKG2A inhibitory receptor

doi: 10.1126/sciadv.adu0690

Figure Lengend Snippet: ( A ) CD16 + , NKG2A + , or NKG2C + population (%) of CD56 + NK cells after antibodies (100 nM) and IL-2 treatment (50 IU/ml) in the coculture system with A549 cells, E:T ratio of 5:1, for 24 hours. ( B ) Left: Graphs showing the cell killing activity of aHER2 × aNKG2A BiNK cotreatment with anti-HER2 IgG1 pertuzumab (ptz IgG1) in A549 and H2030 cells in the presence of primary NK cells. E:T ratio of 2:1. Right: Schematic figure of antibodies treatment schedule. (A) Each symbol represents the value obtained from individual healthy donors. Significance was determined by one-way ANOVA with the Tukey’s post hoc test, * P < 0.05 and ** P < 0.01.

Article Snippet: Primary NK cells were cultured with magnetic-activated cell sorting (MACS) basal NK media with supplementary (Miltenyi Biotec, 130-114-429), 10% (v/v) human serum (Sigma-Aldrich), and human IL-2 (hIL-2) (50 IU/ml; Miltenyi Biotec).

Techniques: Activity Assay