mouse monoclonal cd8a  (fluidigm)

Journal: Frontiers in Immunology

Article Title: TIGIT agonism as a therapeutic strategy to suppress inflammation in hidradenitis suppurativa

doi: 10.3389/fimmu.2026.1761782

Figure Lengend Snippet: (A) Immunohistochemistry images showing PVRL3 staining in normal control skin (NL), hidradenitis suppurativa (HS), and follicular cyst (RFC) samples. (B, C) Quantification of PVRL3 positive cells was performed in QuPath and expressed as the Opercentage of all detected cells. Sample sizes: control (n = 4), HS (n = 52), FC (n = 8). Statistical analysis via nonparametric bootstrap inference. Scale bar = 100 μm. (D) Analysis of publicly available dataset - Log2 fold change comparing HS vs healthy skin - TIGIT and its ligands ( GSE175990 ). Statistical analysis was performed using Wilcoxon rank- sum test with Benjamini-Hochberg false discovery rate. (E) Representative immunofluorescence images of CD8 and TIGIT co-staining in HS and RFC samples. (F) Percentage of CD8+TIGIT+ cells expressed as a proportion of all TIGIT+ cells. (G) Percentage of CD8-TIGIT cells expressed as a proportion of all TIGIT+ cells. CD8 and TIGIT subset analysis was performed in a sample set of HS (n = 5) and RFC (n = 5). Statistical analysis was performed using a nonparametric unpaired t test. Bars represent means. *p<0.05.

Article Snippet: CD8 antibody used CF802079 from Origene.

Techniques: Immunohistochemistry, Staining, Control, Immunofluorescence

Journal: bioRxiv

Article Title: NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses

doi: 10.64898/2025.12.19.695247

Figure Lengend Snippet: Tumor infiltrating lymphocytes (TILs) and single cell suspensions from tumor draining (DLN) and non-tumor draining (NDLN) lymph nodes were isolated at the time of tumor collection from Nlrc5 +/+ and Nlrc5 −/− hosts implanted with B16-V, B16-N-FL or B16-N-SA cells. Cells were counted and stained with fluorochrome conjugated antibodies for lymphoid and NK cell markers and analyzed by flow cytometry using gating strategies depicted in Supplementary Fig. S2. Data from 6-9 mice per group from two independent experiments are shown. (A) Total cellularity (number of CD45 + singlet cells) in TILs, DLN and NDLN. The cell numbers in TILs were equalized to the weight of the tumor mass (TILs per gram; TILS(g -1 )). (B-G) Absolute number of total CD3 + T cells (B), CD4 + T cells (C), CD8 + T cells (D), NK cells (E, NK1.1 + TCRb − ), NKT cells (F, NK1.1 + TCRb + ) and iNKT cells (G, NK1.1 + TCRb + α-GalCer:CD1d + ) were calculated from their percentages within the gated populations (Supplementary Fig. S3). (A-G) Statistics: Mean + SD. Two-way ANOVA with Tukey’s multiple comparison test. * p ≤0.05, ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001. Significant differences between B16-V, B16-N-FL and B16-N-SA are indicated by solid lines. Significant differences between Nlrc5 +/+ and Nlrc5 −/− hosts for the same tumor are indicated by dotted lines.

Article Snippet: To achieve CD8 T cell depletion, anti-CD8a monoclonal antibodies (clone 53-6.7, #BE0004-1, BioXCell) or an isotype control (Rat IgG2a, clone 2A3, #BE0089, BioXCell) were administered intraperitoneally at a dose of 25 μg per mouse on days −1 and +1 relative to tumor implantation.

Techniques: Isolation, Staining, Flow Cytometry, Comparison

Journal: bioRxiv

Article Title: NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses

doi: 10.64898/2025.12.19.695247

Figure Lengend Snippet: TILs and cells from DLN and NDLN from B16-V, B16-N-FL or B16-N-SA tumor bearing Nlrc5 +/+ and Nlrc5 −/− hosts were stained with fluorochrome conjugated antibodies for activation and differentiation markers of T lymphocytes and analyzed by flow cytometry (Supplementary Fig. S2). Data shown are from 6-9 mice per group from two independent experiments. Absolute numbers of CD8 + and CD4 + effector (A,E), effector memory (B, F), central memory (C, G) and naïve (D, H) T cells are shown in TILs (g -1 ), DLN and NDLN. Statistics: Mean + SD. Two-way ANOVA with Tukey’s multiple comparison test. * p ≤0.05, ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001.

Article Snippet: To achieve CD8 T cell depletion, anti-CD8a monoclonal antibodies (clone 53-6.7, #BE0004-1, BioXCell) or an isotype control (Rat IgG2a, clone 2A3, #BE0089, BioXCell) were administered intraperitoneally at a dose of 25 μg per mouse on days −1 and +1 relative to tumor implantation.

Techniques: Staining, Activation Assay, Flow Cytometry, Comparison

Journal: bioRxiv

Article Title: NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses

doi: 10.64898/2025.12.19.695247

Figure Lengend Snippet: (A) Nlrc5 +/+ (A-C) and Nlrc5 −/− (D-F) mice were injected with anti-CD8α, anti-NK1.1 (PK136) or control (IgG2a) antibodies to deplete CD8+ or NK1.1+ cells 24 h prior to and 24 h following subcutaneous implantation of 2 × 10 5 B16-N-FL (A,C,D) or B16-N-SA (B,E,F) cells. Additional doses of Ab were injected 7 and 14 days later. Mice injected with B16-V cells served as additional controls for tumor growth comparison. Data from eight mice per group from two different experiments. (A-C) Growth kinetics (A,B) and tumor weight at sacrifice (C) for B16-N-FL and B16-N-SA tumors in Nlrc5 +/+ hosts. (D-F) Growth kinetics (D,E) and tumor weight at sacrifice (F) for B16-N-FL and B16-N-SA tumors in Nlrc5 −/− hosts. Statistics: Mean + SD. Two-way ANOVA with Tukey’s multiple comparison test. * p≤0.05, *** p≤0.001, **** p≤0.0001.

Article Snippet: To achieve CD8 T cell depletion, anti-CD8a monoclonal antibodies (clone 53-6.7, #BE0004-1, BioXCell) or an isotype control (Rat IgG2a, clone 2A3, #BE0089, BioXCell) were administered intraperitoneally at a dose of 25 μg per mouse on days −1 and +1 relative to tumor implantation.

Techniques: Injection, Control, Comparison

Journal: Oncoimmunology

Article Title: Prostate tumor immune microenvironment changes following immunotherapy shared by patients who developed anti-tumor response or immune-related adverse events

doi: 10.1080/2162402X.2025.2595788

Figure Lengend Snippet: CD8 T cell-rich zones were present in both pre- and post-treatment tumor biopsies. Patient biopsies were collected at 0 weeks (baseline, pre-treatment) and at 12 weeks (post-treatment) and stored as FFPE blocks. Tissue sections (5 µm thick) were obtained and processed for either nCounter (A) or GeoMx analysis (B-D). The whole section was digested to extract RNA and processed following the nCounter workflow, whereas CD8 T cell rich or excluded regions were selected for processing through the GeoMx workflow. Volcano plots resulting from the differential expression analysis are presented here. Gene expression levels between pre- and post-treatment samples were compared using nCounter (A). Protein expression levels in CD8 T cell-rich ROIs between pre- and post-treatment samples were compared using GeoMx (B), and protein expression levels in CD8 T cell-rich regions were Compared to CD8 T cell excluded regions using pre-treatment biopsy samples (C) or in post-treatment biopsy samples using GeoMx (D). The arrows highlight the genes discussed related to the nCounter analysis. p < 0.05 as represented by log 10 ( p -value) >1.3 was considered to be statistically significant and is marked by dashed and dotted green lines in all plots. Fold change of >2 or <–2, as represented by log2(fold change) >1 or <–1 were considered significant changes in expression and are marked by dashed red lines in each plot.

Article Snippet: An anti-human CD8a (Clone ID: OTI3H6) monoclonal antibody (Cat# CF802079 ) was purchased from Origene and conjugated with AF647 using Alexa Fluor 647 antibody labeling kit (Cat# A20186 , Thermo Fisher Scientific) following the manufacturer’s protocol.

Techniques: Quantitative Proteomics, Gene Expression, Expressing

Journal: Oncoimmunology

Article Title: Prostate tumor immune microenvironment changes following immunotherapy shared by patients who developed anti-tumor response or immune-related adverse events

doi: 10.1080/2162402X.2025.2595788

Figure Lengend Snippet: Markers associated with dendritic cells and antigen presentation were associated with the clinical response. Patient biopsies collected pre- and post-treatment were processed as described in . Volcano plots resulting from the differential expression analysis are presented here. Gene expression levels between responders and non-responders were compared using nCounter within pre-treatment biopsy samples (A) or in post-treatment biopsy samples (B). Protein expression levels between responders and non-responders were compared within post-treatment biopsy samples using GeoMx (C), and protein expression levels in CD8 T cell rich regions were compared between responders and non-responders using both pre- and post-treatment biopsy samples (D). The arrows highlight the genes discussed related to the nCounter analysis. p < 0.05 as represented by log 10 ( p -value) >1.3 was considered to be statistically significant and is marked by dashed and dotted green lines in all plots. Fold change of >2 or <−2 as represented by log2(fold change) >1 or <−1 was considered to be a significant change in expression and is marked by dashed red lines in each plot.

Article Snippet: An anti-human CD8a (Clone ID: OTI3H6) monoclonal antibody (Cat# CF802079 ) was purchased from Origene and conjugated with AF647 using Alexa Fluor 647 antibody labeling kit (Cat# A20186 , Thermo Fisher Scientific) following the manufacturer’s protocol.

Techniques: Immunopeptidomics, Quantitative Proteomics, Gene Expression, Expressing

Journal: Oncoimmunology

Article Title: Prostate tumor immune microenvironment changes following immunotherapy shared by patients who developed anti-tumor response or immune-related adverse events

doi: 10.1080/2162402X.2025.2595788

Figure Lengend Snippet: Patients who developed irAEs showed increased T cell activation; whereas PARP expression was elevated in patients that did not develop an irAE. Patient biopsies collected pre- and post-treatment were processed as described in . Volcano plots resulting from the differential expression analysis are presented here. Gene expression levels between patients who experienced an irAE or did not experience irAEs were compared using nCounter within post-treatment biopsy samples (A). Protein expression levels between patients who experienced an irAE or did not experience an irAE were compared within post-treatment biopsy samples using GeoMx (B), and protein expression levels in CD8 T cell rich regions were compared between patients who experienced an irAE or did not experience an irAE within post-treatment biopsy samples (C). The arrows highlight the genes discussed related to the nCounter analysis. p < 0.05 as represented by log 10 ( p -value) >1.3 was considered to be statistically significant and is marked by dashed and dotted green lines in all plots. Fold change of >2 or <−2, as represented by log2(fold change) >1 or <–1 were considered to be a significant change in expression and are marked by dashed red lines in each plot.

Article Snippet: An anti-human CD8a (Clone ID: OTI3H6) monoclonal antibody (Cat# CF802079 ) was purchased from Origene and conjugated with AF647 using Alexa Fluor 647 antibody labeling kit (Cat# A20186 , Thermo Fisher Scientific) following the manufacturer’s protocol.

Techniques: Activation Assay, Expressing, Quantitative Proteomics, Gene Expression