Journal: Nature Communications

Article Title: ERBB2 signaling drives immune cell evasion and resistance against immunotherapy in small cell lung cancer

doi: 10.1038/s41467-025-66800-x

Figure Lengend Snippet: a Schematic of B2M KO generation in murine SCLC primary cell line. Created in BioRender. Meder, L. (2025) https://BioRender.com/4u5k5v3 . b , c Relative MHC-I or PD-L1 expression of WT and MHC-I KO cells analyzed by flow cytometry, determined by mean fluorescence intensity (MFI) normalized to IgG control. Histograms of one representative experiment are shown ( n = 3 biological replicates). d Schematic of experimental setup showing intravenous injection of MHC-I KO and WT cells into immunocompetent C57BL/6/immunodeficient NSG mice, tissue harvest and subsequent IHC and FACS analyses. Created in BioRender. Meder, L. (2025) https://BioRender.com/w6kb2vb . e Representative images of H&E-stained livers and intestinal lymph node tissues from iv. WT and MHC-I KO injected immunocompetent mice. Scale bars, liver 2.5 mm, for lymph nodes 1 mm. Additional FACS based quantification of liver tumor cell infiltration ( n = 4 mice per group). f Representative images of CD45 and CD3 IHC staining in liver tissue from WT and MHC-I KO-injected mice. Quantification was performed using QuPath analysis with 5 regions of interest (ROIs) analyzed per individual ( n = 2 per group). g Representative histograms showing CD107a expression measured by flow cytometry and corresponding quantification (WT n = 4 mice, MHC-I KO n = 5 mice). h Representative images of H&E- and NCAM1-stained liver tissue of MHC-I KO and WT injected immunodeficient NSG mice (MHC-I KO n = 3, WT n = 4) and quantification of tumor cell infiltration in 5 representative ROIs by QuPath analysis. Scale bars, H&E 2.5 mm, NCAM1 100 μm. Statistical analysis was performed using a two-sided, unpaired Student’s t -test. Data in this figure are shown as mean ± SEM. ns not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: The following antibodies and isotype controls were used for staining: CD3 (Alexa-Fluor-700, clone 17A2, Biolegend, Cat. #100216), CD4 (PE-Dazzle 594, clone GK1.5, Biolegend, Cat. # 100456), CD45 (APC-Cy7, clone 30-F11, Biolegend, Cat. # 103116), CTLA-4 (PE, UC10-4B9 Thermo Fisher, Cat. #14-1522-82), CD56 (APC, R&D Systems, clone 809220, Cat. # FAB7820A), CD8a (FITC, clone 53-6.7, Biolegend, Cat. # 100705; Pacific blue, clone 53-6.7, Biolegend, Cat. # 100728), H2Kb (Pacific Blue, clone AF6-88.5, Biolegend, Cat. # 116517), PD-1 (APC, clone 29 F.1A12, Biolegend, Cat. # 135210), PD-L1 (PE-Cy7, clone 10 F.9G2, Biolegend, Cat. # 124313), TIM-3 (PerCP-Cy5.5, clone B8.2C12, Biolegend, Cat. # 134011), Rat IgG2aΚ (Biolegend FITC, Cat. # 400505, PE, Cat. # 400507, PerCP-Cy5.5, Cat. # 400531, APC, Cat. # 400511, Alexa Fluor 700, Cat. # 400528), PE-Dazzle594 Armenian Hamster IgG (PE-Dazzle594, clone HTK888, Biolegend, Cat. # 400951), Rat IgG2bΚ (PE-Cy7, clone RTK4530, Biolegend, Cat. # 400617) and mouse BALB/c IgG2aΚ (Pacific Blue, clone G155-178, BD Biosciences, Cat. # 558118).

Techniques: Expressing, Flow Cytometry, Fluorescence, Control, Injection, Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Conjunctival Intraepithelial Lymphocytes, Lacrimal Cytokines and Ocular Commensal Microbiota: Analysis of the Three Main Players in Allergic Conjunctivitis

doi: 10.3389/fimmu.2022.911022

Figure Lengend Snippet: Description of monoclonal antibodies used.

Article Snippet: , CD56 , APC , MEM-188 , IgG2a, K , Immunostep , 4.

Techniques: Bioprocessing, Marker

Journal: Frontiers in Immunology

Article Title: Conjunctival Intraepithelial Lymphocytes, Lacrimal Cytokines and Ocular Commensal Microbiota: Analysis of the Three Main Players in Allergic Conjunctivitis

doi: 10.3389/fimmu.2022.911022

Figure Lengend Snippet: Lymphoid subset proportions in peripheral blood (PB) and upper tarsal conjunctiva (UTC) in healthy individuals.

Article Snippet: , CD56 , APC , MEM-188 , IgG2a, K , Immunostep , 4.

Techniques:

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were stimulated with or without Ebola VLPs (final concentration: 10 μg/mL of EBOV GP) for 6, 24, 48, 72, and 96 hours. Surface markers and intracellular cytokine expression were assessed by flow cytometry. Strategy for gating single and live CD3 – CD56 + NK cells is shown in . ( A and C ) Representative dot blots showing CD3 – CD56 + NK cells plotted for IFN-γ and TNF-α, respectively. ( B and D ) Summary data from 11 independent experiments performed with PBMCs from 11 (24 and 48 hours) or 5 (6, 72, and 96 hours) independent donors are shown for IFN-γ and TNF-α, respectively. Results are shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, as calculated by 2-tailed paired Student’s t test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Concentration Assay, Expressing, Flow Cytometry

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were stimulated with or without Ebola VLPs as in Figure 2. Cells were stained for the surface and cytotoxicity markers and data acquired and analyzed as described in Methods. Only single and live CD3 – CD56 + NK cells were included in the analysis. Representative dot blots ( A ) and the corresponding summary data from 11 independent experiments performed with PBMCs from 11 (24 and 48 hours) or 5 (6, 72, and 96 hours) independent donors ( B ) showing NK cells’ degranulation (CD107a surface expression) are shown. ( C – F ) Representative histograms for intracellular granzyme B ( C ) and perforin ( E ) expression in gated CD3 – CD56 + NK cells from PBMC cultures stimulated with Ebola VLPs (black) for 48 hours are shown. NK cells from PBMCs left unstimulated are shown in gray. Dotted histograms represent corresponding isotype controls. D (granzyme B) and F (perforin) show corresponding summary data from 8 independent experiments performed with PBMCs from 8 independent donors. ( G ) ADCC killing of target cells mediated by either unexposed (gray circles) or Ebola VLP exposed (black circles) PBMCs. PBMCs exposed to Ebola VLPs for 48 hours were analyzed for their cytotoxic potential by mixing them with the target cells expressing EBOV GP and in the presence of anti-EBOV GP plasma. Combined data from 5 independent experiments with PBMCs from 5 donors are shown. Results are shown as mean ± SEM. ** P < 0.01, *** P < 0.001, as calculated by 2-tailed paired Student’s t test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Staining, Expressing

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were stimulated with or without Ebola VLPs and stained as in and 3. Single and live CD3 – CD56 + NK cells were divided into CD56 bright and CD56 dim NK cells as shown in . Representative dot blots and the corresponding summary data for IFN-γ ( A – D ), TNF-α ( E – H ), and CD107a ( I – L ) from 8 independent experiments with PBMCs from 8 donors are shown. Results are shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 as calculated by 2-tailed paired Student’s t test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Staining

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were left unstimulated or stimulated with Ebola VLPs (10 μg/mL), EBOV GP (10 μg/mL), or EBOV VP40 (5 μg/mL) for 24 hours (IFN-γ and TNF-α) or 48 hours (NK cell degranulation). Cells were stained for the surface and intracellular markers and data acquired as described earlier. Representative dot blots and the corresponding summary data showing gated CD3 – CD56 + NK cells plotted for IFN-γ ( A and B ), TNF-α ( C and D ), and NK cells’ degranulation (CD107a surface expression) ( E and F ) from 6 independent experiments performed with PBMCs from 6 donors are shown. All results are shown as mean ± SEM. ** P < 0.01, *** P < 0.001, as calculated by repeated measures (RM) 1-way ANOVA followed by Dunnett’s multiple comparisons test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Staining, Expressing

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: Purified CD56 + NK cells, cultured alone ( A and B ), with purified CD14 + cells ( C and D ), or with CD14 – CD56 – fraction ( E and F ) and the parent PBMCs ( G and H ) were stimulated with or without EBOV VP40 (5 μg/mL) for 24 hours (IFN-γ) or 48 hours (CD107a). Cells were analyzed flow cytometrically. Representative dot blots with CD3 – CD56 + NK cells plotted for IFN-γ ( A , C , E , and G ) and CD107a ( B , D , F , and H ) are shown. Corresponding summary data from 5 independent experiments performed with cells isolated from 5 donors are shown in I and J , respectively. Results are shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, as calculated by 2-tailed paired Student’s t test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Purification, Cell Culture, Isolation

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were cultured in complete media unstimulated or stimulated with EBOV VP40 (5 μg/mL) for 24 hours. Where indicated blocking monoclonal antibodies for IL-1β, IL-10R, IL-12, IL-15, IL-18, IFN-αβR2 or the corresponding isotype controls were included in the cultures (final concentration of 3 μg/mL). Cells were analyzed flow cytometrically as described earlier. Representative dot blots for CD3 – CD56 + NK cells secreting IFN-γ in the presence of the blocking antibodies that showed statistically significant effect compared with the EBOV VP40 alone are shown ( A ). Corresponding summary data from 5 independent experiments performed with PBMCs from 5 donors are shown ( B ). Results are shown as mean ± SEM. * P < 0.05, ** P < 0.01, as calculated by RM 1-way ANOVA followed by Holm-Šidák multiple comparisons test. IL-10R, IL-10 receptor; IFN-αβR2, IFN-αβ receptor 2.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Cell Culture, Blocking Assay, Concentration Assay

Journal: JCI Insight

Article Title: Ebola virus protein VP40 stimulates IL-12– and IL-18–dependent activation of human natural killer cells

doi: 10.1172/jci.insight.158902

Figure Lengend Snippet: PBMCs were cultured, then stimulated with EBOV VP40 (5 μg/mL) for 48 hours in the presence of blocking monoclonal antibodies for IL-1β, IL-10R, IL-12, IL-15, IL-18, and IFN-αβR2 or the corresponding isotype controls (all at a final concentration of 3 μg/mL) and analyzed flow cytometrically as described earlier. Representative dot blots with CD3 – CD56 + NK cells plotted for surface CD107a are shown ( A ). Corresponding summary data from 5 independent experiments performed with PBMCs from 5 donors are shown ( B ). Results are shown as mean ± SEM. * P < 0.05, ** P < 0.01, as calculated by RM 1-way ANOVA followed by Holm-Šidák multiple comparisons test.

Article Snippet: Purity of the isolated cell populations was assessed by flow cytometry using APC-conjugated mouse anti-human CD56 (clone CMSSB, MBL International) and V500-conjugated mouse anti-human CD14 (clone M5E2, BD Biosciences) antibodies.

Techniques: Cell Culture, Blocking Assay, Concentration Assay