Journal: iScience

Article Title: PD-L1 + CD8 + T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance

doi: 10.1016/j.isci.2022.103785

Figure Lengend Snippet:

Article Snippet: anti-human CD57 -176Yb , Fluidigm , Cat# 3176019B.

Techniques: Recombinant, Antibody Labeling, Enzyme-linked Immunosorbent Assay, Sequencing, Software

Journal: Clinical and Experimental Immunology

Article Title: Phenotypic and functional characteristics of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected individuals

doi: 10.1093/cei/uxac082

Figure Lengend Snippet: Expansion of highly differentiated CD57 + NKG2C + NK cells in donors infected with HIV-1, HCMV, or both viruses. Co-expression of CD57 and NKG2C on NK cells from 54 HIV-1-infected and 18 HIV-1-uninfected donors, with and without HCMV infection, was assessed by flow cytometry. ( A ) Gating strategy used to identify the expression of CD57 and NKG2C on CD56 dim NK cells in three representative HIV-1-infected donors. Two donors had undetermined HCMV serostatuses and one was HCMV seropositive. The graphs depict the percentage of CD56 dim NK cells that co-express CD57 and NKG2C in ( B ) HIV-1-infected and HIV-1-uninfected subjects, regardless of HCMV infection, ( C ) HIV-1-uninfected subjects with and without HCMV infection and ( D ) HCMV-infected subjects with and without HIV-1 co-infection. Donors with confirmed HCMV seropositivity are depicted in red. Donors with suppressed viremia are represented with circles, and those with detectable viremia are represented with triangles. Data were compared with Mann–Whitney tests. P < 0.05 was considered significant. Columns represent medians.

Article Snippet: The efficacy of the depletion was assessed by flow cytometry using CD3 PerCP (clone SK7; BD Biosciences), CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences) and CD57 FITC (clone TB03; Miltenyi Biotec) antibodies.

Techniques: Infection, Expressing, Flow Cytometry, MANN-WHITNEY

Journal: Clinical and Experimental Immunology

Article Title: Phenotypic and functional characteristics of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected individuals

doi: 10.1093/cei/uxac082

Figure Lengend Snippet: Impact of ART on the frequency of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected subjects. The frequency of highly differentiated NK cells in pre- and post-ART samples from 10 subjects infected with both HIV-1 and HCMV was evaluated by flow cytometry. Data were compared with a Wilcoxon matched-pairs signed rank test. P < 0.05 was considered significant.

Article Snippet: The efficacy of the depletion was assessed by flow cytometry using CD3 PerCP (clone SK7; BD Biosciences), CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences) and CD57 FITC (clone TB03; Miltenyi Biotec) antibodies.

Techniques: Infection, Flow Cytometry

Journal: Clinical and Experimental Immunology

Article Title: Phenotypic and functional characteristics of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected individuals

doi: 10.1093/cei/uxac082

Figure Lengend Snippet: Functional characteristics of highly differentiated CD57 + NKG2C + NK cells from HIV-1-infected donors. NK cell activation was measured by flow cytometry as the percentage of CD56 dim NK cells expressing IFNγ after antibody-dependent stimulation. ( A ) FACS plots showing the gating on IFNγ + CD56 dim NK cells in PBMC incubated 5 hours in the absence of target cells, with HIV-1 BaL gp120-coated CEM.NKr-CCR5 target cells alone (no antibody control), 721.221 target cells alone (no antibody control), gp120-coated CEM.NKr-CCR5 cells in the presence of pooled HIV-1 immunoglobulin (HIVIG) or 721.221 cells pre-coated with the anti-CD20 antibody Rituximab (RTX). ( B ) The graphs depict the percentage of IFNγ + cells within CD56 dim CD57 - , CD56 dim CD57 + NKG2C - and CD56 dim CD57 + NKG2C + NK cells stimulated with CEM.NKr-CCR5 cells with HIVIG (left) or 721.221 cells with RTX (right) for 12 HIV-1-infected donors. Donors with confirmed HCMV seropositivity are depicted in red. Donors with suppressed viremia are represented with circles, and those with detectable viremia are represented with triangles. Data were compared with Friedman tests followed by Dunn’s post-tests to assess differences between CD56 dim CD57 + NKG2C + cells and CD56 dim CD57 - or CD56 dim CD57 + NKG2C - NK cells. P < 0.05 was considered significant.

Article Snippet: The efficacy of the depletion was assessed by flow cytometry using CD3 PerCP (clone SK7; BD Biosciences), CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences) and CD57 FITC (clone TB03; Miltenyi Biotec) antibodies.

Techniques: Functional Assay, Infection, Activation Assay, Flow Cytometry, Expressing, Incubation, Control

Journal: Clinical and Experimental Immunology

Article Title: Phenotypic and functional characteristics of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected individuals

doi: 10.1093/cei/uxac082

Figure Lengend Snippet: NKG2C-mediated redirected lysis of target cells by highly differentiated NK cells. PBMC from HIV-1-infected donors were incubated with Fc-receptor-bearing P815 target cells for four hours in the absence or presence of anti-NKG2A, anti-NKG2C or anti-CD16 antibodies. ( A ) The graph depicts the redirected lysis of P815 cells induced by the engagement of the inhibitory receptor NKG2A or the activating receptors NKG2C or CD16. Columns represent medians. ( B ) The correlation between NKG2C-mediated redirected lysis and co-expression of CD57 and NKG2C on CD56 dim NK cells is shown in the graph. Data were analyzed using a non-parametric Spearman correlation. P < 0.05 was considered significant. ( C ) PBMC from six HIV-1-infected donors were depleted of CD57 + cells. The FACS plots depict the percentage of CD57 + NK cells within the CD56 dim NK cell population before and after CD57 depletion in a representative donor. ( D ) The graph shows the percentage of CD57 + NK cells within the CD56 dim NK cell population pre- and post-CD57-depletion in all six donors. ( E ) The graph depicts the redirected lysis of P815 cells through NKG2C in whole PBMC and CD57-depleted PBMC. ( F ) NK cells were enriched from PBMC from six HIV-1-infected donors. The graph shows the percentage of NK cells within the lymphocyte population pre- and post-enrichment. ( G ) The graph depicts the redirected lysis of P815 target cells by enriched NK cells in the absence of antibody or the presence of anti-NKG2C antibody. Donors with confirmed HCMV seropositivity are depicted in red. All donors had suppressed viremia. Data were compared with Wilcoxon matched-pairs signed rank tests. P < 0.05 was considered significant.

Article Snippet: The efficacy of the depletion was assessed by flow cytometry using CD3 PerCP (clone SK7; BD Biosciences), CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences) and CD57 FITC (clone TB03; Miltenyi Biotec) antibodies.

Techniques: Lysis, Infection, Incubation, Expressing

Journal: Clinical and Experimental Immunology

Article Title: Phenotypic and functional characteristics of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected individuals

doi: 10.1093/cei/uxac082

Figure Lengend Snippet: Phenotypic characterization of highly differentiated CD57 + NKG2C + NK cells in HIV-1-infected donors. Flow cytometry was used to assess the expression of NKG2A on CD56 dim NK cells that were CD57 - , CD57 + NKG2C - and CD57 + NKG2C + . ( A ) The graph depicts the percentage of NKG2A + NK cells within CD57 - and CD57 + NKG2C +/- CD56 dim NK cells. Columns represent medians. Data were compared with a Friedman test followed by Dunn’s post-tests, which assessed differences between CD57 + NKG2C + and CD57 - or CD57 + NKG2C - NK cells. ( B ) The graph shows the correlation between NKG2A expression and CD57/NKG2C co-expression on CD56 dim NK cells. Donors with confirmed HCMV seropositivity are depicted in red. Donors with suppressed viremia are represented with circles, and those with detectable viremia are represented with triangles. Data were analyzed using a non-parametric Spearman correlation. P < 0.05 was considered significant.

Article Snippet: The efficacy of the depletion was assessed by flow cytometry using CD3 PerCP (clone SK7; BD Biosciences), CD56 PE-Cy7 (clone NCAM16.2; BD Biosciences) and CD57 FITC (clone TB03; Miltenyi Biotec) antibodies.

Techniques: Infection, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: 29-Color Flow Cytometry: Unraveling Human Liver NK Cell Repertoire Diversity

doi: 10.3389/fimmu.2019.02692

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: CD57 , TB03 , APC-Vio770 , , 780/60 , 50 , No , Miltenyi Biotec , Maturation.

Techniques: Marker, Staining

Journal: NPJ Precision Oncology

Article Title: CD57-positive CD8 + T cells define the response to anti-programmed cell death protein-1 immunotherapy in patients with advanced non-small cell lung cancer

doi: 10.1038/s41698-024-00513-0

Figure Lengend Snippet: a – c Flow cytometry-based quantification of T cells, CD8 + T cells, CD57 + CD8 + T cells and CD57 - CD8 + T cells in the prospective NSCLC cohorts. d CD57 + CD8 + T cells/T cells ratio ( p < 0.001), e CD57 + CD8 + T cells/ CD8 + T cells ratio ( p = 0.001) and f total CD57 + T cells/T cells ( p = 0.001) as determined by flow cytometry predicted clinical response to immunotherapy ( n = 27). g Total CD8 + T cells/T cells ratio was not of predictive value ( p = 0.059, n = 27). h ROC curve analyzed the ability of CD57 + CD8 + T cells/T cells ratio to identify responders (AUC = 0.9375, n = 27). Sensitivity (87.5%) refers to the proportion of true positive subjects with the disease among all subjects with disease. Specificity (100.0%) refers to the proportion of true negative subjects without the disease among subjects without disease. PPV (84.6%) refers to the proportion of patients with positive results among subjects with positive results. NPV (100.0%) refers to the proportion of subjects without disease with a negative result among subjects with negative results. Accuracy (92.6%) refers to the proportion of subjects correctly classified among all subjects. i DCB proportion comparing the ability of CD57 + CD8 + T cells/T cells ratio as determined by flow cytometry, total CD8 + T cells/T cells ratio by flow cytometry, and PD-L1 status by conventional IHC to predict treatment response. j ROC curves for predicting treatment response using the CD57 + CD8 + T cells/T cells ratio (AUC = 0.733), CD8 + T cells/T cells ratio (AUC = 0.631), and PD-L1 positivity (AUC = 0.560) ( n = 48). *** p ≤ 0.001. NSCLC non-small cell lung cancer, ROC receiver operating characteristic, AUC area under the curve, PPV positive predictive value, NPV negative predictive value, DCB durable clinical benefit, PD-L1 programmed death-ligand 1, IHC immunohistochemistry.

Article Snippet: Briefly, each FFPE tissue section was baked at 65 °C for 1 h. After deparaffinization, rehydration, and microwave antigen repair, the slides were blocked (Akoya Biosciences, USA), and incubated with primary antibodies against CD3 (D7A6E, CST, 85061 T, 1:200), CD57 (HNK-1, CST, 72031 S, 1:200), and CD8 (EPR22483-288, Abcam, ab245118, 1:500), respectively, followed by incubation with Opal Polymer HRP Ms+Rb (Akoya Biosciences, USA).

Techniques: Flow Cytometry, Immunohistochemistry

Journal: NPJ Precision Oncology

Article Title: CD57-positive CD8 + T cells define the response to anti-programmed cell death protein-1 immunotherapy in patients with advanced non-small cell lung cancer

doi: 10.1038/s41698-024-00513-0

Figure Lengend Snippet: a Representative images of NSCLC tissue stained using mIHC/IF [CD3 (green), CD8 (red), CD57 (pink), DAPI (blue)]. b mIHC/IF-based CD57 + CD8 + T cells/T cells ratio ( p < 0.001) and ( c ) CD57 + CD8 + T cells/CD8 + T cells ratio ( p < 0.001) predicted immunotherapeutic response in a retrospective NSCLC cohort ( n = 90). d Total CD8 + T cells/T cells ratio by mIHC/IF was not of predictive value to immunotherapy ( p = 0.679, n = 90). e Spearman’s rank correlation analysis was performed to compare the CD57 + CD8 + T cells/T cells ratio ( r = 0.644, p < 0.001), and f CD8 + T cells/T cells ratio ( r = 0.346, p = 0.016) in blood and tumor samples ( n = 48). *** p < 0.001. mIHC/IF multiplex immunohistochemistry/immunofluorescence, NSCLC non-small cell lung cancer.

Article Snippet: Briefly, each FFPE tissue section was baked at 65 °C for 1 h. After deparaffinization, rehydration, and microwave antigen repair, the slides were blocked (Akoya Biosciences, USA), and incubated with primary antibodies against CD3 (D7A6E, CST, 85061 T, 1:200), CD57 (HNK-1, CST, 72031 S, 1:200), and CD8 (EPR22483-288, Abcam, ab245118, 1:500), respectively, followed by incubation with Opal Polymer HRP Ms+Rb (Akoya Biosciences, USA).

Techniques: Staining, Multiplex Assay, Immunohistochemistry, Immunofluorescence

Journal: NPJ Precision Oncology

Article Title: CD57-positive CD8 + T cells define the response to anti-programmed cell death protein-1 immunotherapy in patients with advanced non-small cell lung cancer

doi: 10.1038/s41698-024-00513-0

Figure Lengend Snippet: a Volcano plot showing the 475 DEGs between CD57 + CD8 + T cells and CD57 − CD8 + T cells, including 133 upregulated genes and 342 downregulated genes. Red and blue colors represent upregulated and downregulated genes, respectively. b Clustering analysis of DEGs and samples. The color scale bar for heat intensity indicates Log2(Fold Change). Columns, samples; rows, DEGs. The samples were grouped into two distinct clusters: CD57 + CD8 + T cell cluster and CD57 - CD8 + T cell cluster. c GO analysis of DEGs. The most enriched 30 GO terms in biological process, cellular component, and molecular function. The y axis represents GO terms and the x axis represents the value of -log10 ( p -value). d KEGG enrichment analysis of DEGs. The x axis represents enrichment score and the y axis represents pathway. Size and color of the bubble represent the amount of DEGs enriched in pathway and enrichment significance, respectively. e Representative GSEA results showing enrichment of the immune-associated pathways in CD57 + CD8 + T cells. DEGs differentially expressed genes, GO Gene Ontology, KEGG Kyoto Encyclopedia of Genes and Genomes, GSEA Gene Set Enrichment Analysis, NSCLC non-small cell lung cancer.

Article Snippet: Briefly, each FFPE tissue section was baked at 65 °C for 1 h. After deparaffinization, rehydration, and microwave antigen repair, the slides were blocked (Akoya Biosciences, USA), and incubated with primary antibodies against CD3 (D7A6E, CST, 85061 T, 1:200), CD57 (HNK-1, CST, 72031 S, 1:200), and CD8 (EPR22483-288, Abcam, ab245118, 1:500), respectively, followed by incubation with Opal Polymer HRP Ms+Rb (Akoya Biosciences, USA).

Techniques:

Journal: NPJ Precision Oncology

Article Title: CD57-positive CD8 + T cells define the response to anti-programmed cell death protein-1 immunotherapy in patients with advanced non-small cell lung cancer

doi: 10.1038/s41698-024-00513-0

Figure Lengend Snippet: Results of binomial logistic regression analysis to stratify DCB and NDB patients.

Article Snippet: Briefly, each FFPE tissue section was baked at 65 °C for 1 h. After deparaffinization, rehydration, and microwave antigen repair, the slides were blocked (Akoya Biosciences, USA), and incubated with primary antibodies against CD3 (D7A6E, CST, 85061 T, 1:200), CD57 (HNK-1, CST, 72031 S, 1:200), and CD8 (EPR22483-288, Abcam, ab245118, 1:500), respectively, followed by incubation with Opal Polymer HRP Ms+Rb (Akoya Biosciences, USA).

Techniques: