Journal: Frontiers in Immunology

Article Title: Downregulation of HLA Class I Renders Inflammatory Neutrophils More Susceptible to NK Cell-Induced Apoptosis

doi: 10.3389/fimmu.2019.02444

Figure Lengend Snippet: NKR ligand expression on in vitro -activated neutrophils. (A) Surface expression of HLA-ABC, CD11b, CD66 or CD62L as indicated, on resting neutrophils kept on ice (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C alone or in presence of the TLR-agonist CL097 and GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). (B) Surface expression of indicated ligands to NKRs on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C with CL097/GM-CSF (paired t -test). (C) Expression of NKR ligands and neutrophil degranulation markers on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 1, 2, or 4 h at 37°C with CL097/GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). Error bars represent SEM.

Article Snippet: Following mAbs were used for detection of surface markers: anti-CD11b APC (clone: ICRF44), CD48 BV421 (TÜ145), HLA-ABC FITC (G46-2.6), CD62L APC (DREG-56), CD66 PE (B1.1; all from BD Biosciences, CA, USA), HLA-C (DT9, Merck Millipore), HLA-ABC Alexa Fluor 647 (W6/32) and HLA-E PE-Cy7 (3D12; both from Biolegend), B7-H6 (875002), ULBP1 PerCP (170818), ULBP-3 PE (166510), ULBP-2/5/6 APC (165903; all from R&D Systems, MN, USA), CD56 PE-Cy7 (N901, Beckman Coulter, IN, USA), MICA/B viobright FITC (6D4, Miltenyi Biotech, Bergisch Gladbach, Germany), and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, CA, USA).

Techniques: Expressing, In Vitro

Journal: Frontiers in Immunology

Article Title: Downregulation of HLA Class I Renders Inflammatory Neutrophils More Susceptible to NK Cell-Induced Apoptosis

doi: 10.3389/fimmu.2019.02444

Figure Lengend Snippet: NKR ligand expression on in vitro -activated neutrophils. (A) Surface expression of HLA-ABC, CD11b, CD66 or CD62L as indicated, on resting neutrophils kept on ice (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C alone or in presence of the TLR-agonist CL097 and GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). (B) Surface expression of indicated ligands to NKRs on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C with CL097/GM-CSF (paired t -test). (C) Expression of NKR ligands and neutrophil degranulation markers on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 1, 2, or 4 h at 37°C with CL097/GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). Error bars represent SEM.

Article Snippet: Following mAbs were used for detection of surface markers: anti-CD11b APC (clone: ICRF44), CD48 BV421 (TÜ145), HLA-ABC FITC (G46-2.6), CD62L APC (DREG-56), CD66 PE (B1.1; all from BD Biosciences, CA, USA), HLA-C (DT9, Merck Millipore), HLA-ABC Alexa Fluor 647 (W6/32) and HLA-E PE-Cy7 (3D12; both from Biolegend), B7-H6 (875002), ULBP1 PerCP (170818), ULBP-3 PE (166510), ULBP-2/5/6 APC (165903; all from R&D Systems, MN, USA), CD56 PE-Cy7 (N901, Beckman Coulter, IN, USA), MICA/B viobright FITC (6D4, Miltenyi Biotech, Bergisch Gladbach, Germany), and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, CA, USA).

Techniques: Expressing, In Vitro

Journal: Frontiers in Immunology

Article Title: Downregulation of HLA Class I Renders Inflammatory Neutrophils More Susceptible to NK Cell-Induced Apoptosis

doi: 10.3389/fimmu.2019.02444

Figure Lengend Snippet: NKR ligand expression on in vitro -activated neutrophils. (A) Surface expression of HLA-ABC, CD11b, CD66 or CD62L as indicated, on resting neutrophils kept on ice (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C alone or in presence of the TLR-agonist CL097 and GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). (B) Surface expression of indicated ligands to NKRs on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C with CL097/GM-CSF (paired t -test). (C) Expression of NKR ligands and neutrophil degranulation markers on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 1, 2, or 4 h at 37°C with CL097/GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). Error bars represent SEM.

Article Snippet: Following mAbs were used for detection of surface markers: anti-CD11b APC (clone: ICRF44), CD48 BV421 (TÜ145), HLA-ABC FITC (G46-2.6), CD62L APC (DREG-56), CD66 PE (B1.1; all from BD Biosciences, CA, USA), HLA-C (DT9, Merck Millipore), HLA-ABC Alexa Fluor 647 (W6/32) and HLA-E PE-Cy7 (3D12; both from Biolegend), B7-H6 (875002), ULBP1 PerCP (170818), ULBP-3 PE (166510), ULBP-2/5/6 APC (165903; all from R&D Systems, MN, USA), CD56 PE-Cy7 (N901, Beckman Coulter, IN, USA), MICA/B viobright FITC (6D4, Miltenyi Biotech, Bergisch Gladbach, Germany), and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, CA, USA).

Techniques: Expressing, In Vitro

Journal: Frontiers in Immunology

Article Title: Downregulation of HLA Class I Renders Inflammatory Neutrophils More Susceptible to NK Cell-Induced Apoptosis

doi: 10.3389/fimmu.2019.02444

Figure Lengend Snippet: Decrease of surface HLA class I molecules is not explained by internalization. Graph shows resting neutrophils stained with FITC-conjugated HLA-ABC antibody prior to addition of buffer (ctrl) or CL097/GM-CSF for 20 min at 37°C, after which FITC-quenching antibody was added.

Article Snippet: Following mAbs were used for detection of surface markers: anti-CD11b APC (clone: ICRF44), CD48 BV421 (TÜ145), HLA-ABC FITC (G46-2.6), CD62L APC (DREG-56), CD66 PE (B1.1; all from BD Biosciences, CA, USA), HLA-C (DT9, Merck Millipore), HLA-ABC Alexa Fluor 647 (W6/32) and HLA-E PE-Cy7 (3D12; both from Biolegend), B7-H6 (875002), ULBP1 PerCP (170818), ULBP-3 PE (166510), ULBP-2/5/6 APC (165903; all from R&D Systems, MN, USA), CD56 PE-Cy7 (N901, Beckman Coulter, IN, USA), MICA/B viobright FITC (6D4, Miltenyi Biotech, Bergisch Gladbach, Germany), and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, CA, USA).

Techniques: Staining

Journal: Frontiers in Immunology

Article Title: Downregulation of HLA Class I Renders Inflammatory Neutrophils More Susceptible to NK Cell-Induced Apoptosis

doi: 10.3389/fimmu.2019.02444

Figure Lengend Snippet: NKR ligand expression on in vitro -activated neutrophils. (A) Surface expression of HLA-ABC, CD11b, CD66 or CD62L as indicated, on resting neutrophils kept on ice (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C alone or in presence of the TLR-agonist CL097 and GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). (B) Surface expression of indicated ligands to NKRs on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 20 min at 37°C with CL097/GM-CSF (paired t -test). (C) Expression of NKR ligands and neutrophil degranulation markers on resting neutrophils (ctrl) or in vitro -activated neutrophils stimulated for 1, 2, or 4 h at 37°C with CL097/GM-CSF (one-way ANOVA followed by Dunnett's multiple comparisons test). Error bars represent SEM.

Article Snippet: Following mAbs were used for detection of surface markers: anti-CD11b APC (clone: ICRF44), CD48 BV421 (TÜ145), HLA-ABC FITC (G46-2.6), CD62L APC (DREG-56), CD66 PE (B1.1; all from BD Biosciences, CA, USA), HLA-C (DT9, Merck Millipore), HLA-ABC Alexa Fluor 647 (W6/32) and HLA-E PE-Cy7 (3D12; both from Biolegend), B7-H6 (875002), ULBP1 PerCP (170818), ULBP-3 PE (166510), ULBP-2/5/6 APC (165903; all from R&D Systems, MN, USA), CD56 PE-Cy7 (N901, Beckman Coulter, IN, USA), MICA/B viobright FITC (6D4, Miltenyi Biotech, Bergisch Gladbach, Germany), and Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen, CA, USA).

Techniques: Expressing, In Vitro