cd8 apc cy7  (Cytek Biosciences)

Journal: bioRxiv

Article Title: Immune and Mutational Profile of Gene-Edited Low-Immunogenic Human Primary Cholangiocyte Organoids

doi: 10.1101/2025.01.20.628680

Figure Lengend Snippet: (a) Graphs showing percentage of positive mouse and human CD45 cells, percentage of human CD3, CD8, CD19 and human CD45 cell counts in weekly in peripheral blood (tail vein bleeds) for the two PBMC donor groups (PBMC Donor A, top; PBMC Donor B, bottom). Error bars represent mean±SEM of 4-8 mice per group.

Article Snippet: Humanization was evaluated weekly by flow cytometry of tail-vein bleeds collected in heparin-coated tubes (Sarstedt Ltd, 20.1309), followed by red cell lysis removal (StemCell Technologies, 07850) according to manufacturer’s instructions and using the following conjugated antibodies: mouse anti-human CD45-FITC (1:50, Thermo Fisher Scientific, 11-0459-42, clone [HI30]), mouse anti-mouse CD45.1-PE-Cy7 (1:100, Thermo Fisher Scientific, 25-0453-82, clone [A20]), mouse anti-human CD3-APC (1:50, Biolegend, clone [UCHT1]), mouse anti-human CD19-PE (1:50, Biolegend, 363004, clone [SJ25C1]), and mouse anti-human CD8-APC/Cy7 (1:50, Thermo Fisher Scientific, A15441, clone [RFT8]).

Techniques:

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Antibodies used for flow cytometry analysis and sorting of T cell populations from tumor and control bladder tissues.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques: Flow Cytometry, Staining

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Gating strategy for analysis of Tregs, Th cells, and CD8 + T cells ( A ). Immune cell infiltration measured as CD45 + out of all viable cells ( B ). Frequency of T cells and T cell subsets out of all CD45 + cells ( C – G ). Treg = regulatory T cells, Th = T helper, NMIBC = non-muscle-invasive bladder cancer, MIBC = muscle-invasive bladder cancer. Statistical significance was determined by the Kruskal–Wallis test followed by the two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli post-test.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques:

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Infiltration of CD45 + immune cells out of all viable cells and frequency of T cell populations out of CD45 + immune cells in tumor samples according to sex ( A – D , F , G ). Ratio of CD4 + /CD8 + T cells ( E ) Statistical significance was determined by Kruskal–Wallis test followed by two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli post-test.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques:

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Frequency of immune cells and T cell populations in tumor samples classified according to the Lund Taxonomy molecular subtypes. Infiltration of CD45 + immune cells among all viable cells ( A ). Immune infiltration in MIBC samples only ( B ). Percentages of T cells, CD4 + T cells, and CD8 + T cells out of all CD45 + cells ( C – E ). Ratio of CD4 + /CD8 + T cells ( F ). Percentages of Tregs and Th cells out of CD45 + cells ( G , H ). Statistical significance was determined through the Kruskal–Wallis test followed by the two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli post-test.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques:

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: T cell populations in tumor samples classified according to the Lund Taxonomy molecular subtypes. Percentage of T cells ( A ), CD8 + T cells ( B ), CD4 + T cells ( C ), Tregs ( D ), and Th ( E ) out of all viable cells. Statistical significance was determined by Kruskal–Wallis test followed by two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli post-test.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques:

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Transcriptional analysis of CD8 + T cells from non-malignant bladder tissue and from GU and Uro tumor biopsies. Principal component analysis ( A ). Heatmap of differentially expressed genes between CD8 + T cells from GU and Uro subtypes ( B ). GSEA using an exhaustion profile (Tirosh et al.’s 28-gene exhaustion profile) ( C ). Volcano plot with the differentially expressed genes (GU vs. control) marked in red, with the top 40 annotated ( D ). Volcano plot with the differentially expressed genes (Uro vs. control) marked in red, with the top 40 annotated ( E ). Percentage of CD8 + cells expressing the exhaustion markers CTLA-4, LAG3, PD-1, TIGIT, or TIM-3 measured by flow cytometry ( F ). Statistical significance assessed by Mann–Whitney U test.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques: Expressing, Flow Cytometry, MANN-WHITNEY

Journal: Cells

Article Title: Distinct Infiltration of T Cell Populations in Bladder Cancer Molecular Subtypes

doi: 10.3390/cells13110926

Figure Lengend Snippet: Pathway analysis of differentially expressed genes between CD8 + T cells from GU subtype and the control ( A ) and Uro subtype and the controls ( B ). padj = 0.05 marked in red.

Article Snippet: APC-Cy7 Mouse Anti-Human CD8 , SK1 , BD biosciences.

Techniques:

Journal: ImmunoHorizons

Article Title: A novel humanized PD-1/PD-L1 mouse model permits direct comparison of antitumor immunity generated by FDA-approved PD-1 and PD-L1 inhibitors

doi: 10.4049/immunohorizons.2200054

Figure Lengend Snippet: (A) Schematic of experimental timeline. (B, C) Cells from spleen were isolated and T cell marker expression was captured via CyTOF. High dimensional data was analyzed via Cytobank software’s viSNE algorithm. CD3+CD8+ T cells from representative PBS and Avelumab samples are shown (n=3 PBS, n=3 Avelumab analyzed total). Density gradient indicates a population more prominent in the Avelumab-treated mice (pink arrows). Additional plots display relative expression of various T cell markers. (D) In a separate analysis, CITRUS using SAM with a 5% FDR was completed using all CD3+CD8+ T cells to detect differences in the abundance of T cell subsets in PBS vs. Avelu treated mice. Parent cluster #119994 (highlighted red, with progeny clusters shaded in gray) had a significantly higher abundance in Avelumab-treated mice. (E) Cluster abundance shown as boxplot (n=6 separate CITRUS with SAM repeats produced similar results). (F) CITRUS tree with expression of markers overlayed, indicates high expression (red) to low expression (blue) of each marker. Gray shading shows parent cluster #119994 and progeny, as in D. (G) Flow cytometry completed on separate samples confirms CyTOF findings (n= 7 mice/group, experimental timeline as in A). Bars, mean; error bars, s.e.m. (*P < 0.05). (H) Above, schematic of experimental timeline. Below, average tumor growth curves for each group (n=6 in Avelu group; n=5 Avelu + anti-CD8 treatment group, dots are mean; error bars are s.e.m.).

Article Snippet: The antibodies used included anti-mouse CD8-BV421 (clone 53–6.7), anti-mouse CD11a-PerCp/Cy5.5 (clone M17/4), anti-mouse CX3CR1-PE/Cy7 (clone SA011F11), anti-mouse PD-1-APC (clone RMPI30), anti-human PD-1-APC (clone EH12.2H7), anti-mouse CD11c-BV421 (clone N418), anti-mouse CD4-FITC (RM4–5), anti-mouse CD44-BV711 (IM7), anti-mouse CD45R/B220-BV605 (RA3–6B2), and anti-mouse ICOS-APC (C398.4A), all from BioLegend; iTAg MHC tetramer H-2Kb OVA, SIINFEKL-PE from MBL; anti-mouse B220-SupreBright 600 (RA3–6B2), anti-mouse CD4-eFluor450 (RM4–5), and anti-human PD-L1-PE (clone M1H1), all from Invitrogen; anti-mouse/rat FOXP3-FITC (FJK-16s) from eBioscience; anti-mouse CXCR5-biotin (2G8) from BD Biosciences; and anti-mouse CD8-APC-Cy7 (53-6.7) and anti-CD62L-PE-Cy7 (MEL-14) from TONBO Biosciences.

Techniques: Isolation, Marker, Expressing, Produced, Flow Cytometry

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Primary surface antibody staining panel: Pt25 multilineage panel

Article Snippet: Mouse monoclonal anti-Human CD8 (clone SK1) APC-Cy7 , BD Biosciences , Cat#561945; RRID: AB_396892.

Techniques: Staining

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Gating strategy for multilineage chimerism, as well as donor and recipient T cell phenotypes, in post-transplant ileal biopsies, using ITx Pt25 POD241 IEL as an example (A) Representative contour plots depicting gating strategy of HLA-ABC + CD45 + cells, CD3 + T cells, CD3 + γδTCR + T cells, CD3 + γδTCR − CD4 + T cells, CD3 + γδTCR − CD8 + T cells, CD19 + B cells, and CD3 - CD56 + NK cells. (B) Percentages of donor (“D”) and recipient (“R”) multilineage chimerism in Pt25 POD241 IELs. Pt25’s donor is HLA-B12 + . Pt25, the recipient, is HLA-A68 + , which has serological cross-reactivity with HLA-A28. (C) Gating strategy for donor (“D”) and recipient (“R”) CD4 + and CD8 + T cells in Pt25 POD241 IELs. CD45RA vs. CCR7 to categorize naïve T cells (CD45RA + CCR7 + ), central memory T cells (TCMs: CD45RA − CCR7 + ), effector memory T cells (TEMs: CD45RA − CCR7 − ), and terminally-differentiated effector memory T cells (TEMRAs: CD45RA + CCR7 - ), tissue-resident memory T cells (TRMs: CD69 + CD103 +/− CD49a +/ − ), effector T cells (CD28 + NKG2D +/− ).

Article Snippet: Mouse monoclonal anti-Human CD8 (clone SK1) APC-Cy7 , BD Biosciences , Cat#561945; RRID: AB_396892.

Techniques:

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet: Primary surface antibody staining panel: Pt25 B-cell panel

Article Snippet: Mouse monoclonal anti-Human CD8 (clone SK1) APC-Cy7 , BD Biosciences , Cat#561945; RRID: AB_396892.

Techniques: Staining

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Human CD8 (clone SK1) APC-Cy7 , BD Biosciences , Cat#561945; RRID: AB_396892.

Techniques: Recombinant, Software, Flow Cytometry

Journal: STAR Protocols

Article Title: Chimerism and phenotypic analysis of intraepithelial and lamina propria T cells isolated from human ileal biopsies after intestinal transplantation

doi: 10.1016/j.xpro.2023.102192

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-Human CD8 (clone SK1) APC-Cy7 , BD Biosciences , Cat#561945; RRID: AB_396892.

Techniques: Concentration Assay, Staining