Journal: Cell Reports Medicine

Article Title: T-bet+ CXCR3+ B cells drive hyperreactive B-T cell interactions in multiple sclerosis

doi: 10.1016/j.xcrm.2025.102027

Figure Lengend Snippet:

Article Snippet: Mouse anti-CD278 (ICOS) (clone ISA-3); (FACS, in vitro ) , Thermo Fisher Scientific , Cat# 17-9948-41; RRID: AB_10597585.

Techniques: In Vitro, Control, Recombinant, Staining, Purification, Electron Microscopy, Lysis, Ubiquitin Proteomics, Cell Isolation, Enzyme-linked Immunosorbent Assay, Isolation, Multiplex Assay, Quantitation Assay, Modification, Library Quantification, RNA Sequencing, Sequencing, Software, Cell Culture, Sterility

Journal: Cancer Discovery

Article Title: Temporal Genomic Analysis of Homogeneous Tumor Models Reveals Key Regulators of Immune Evasion in Melanoma

doi: 10.1158/2159-8290.CD-23-1422

Figure Lengend Snippet: Opal and CODEX multiplexed tissue imaging data validate increased M2-like macrophage infiltration and T cell exhaustion in the nonrejected SCCs. A, Representative Opal Multiplex IHC stains for F4/80 (red), CD204 (yellow), and CD206 (green) in tumors derived from SCC31, SCC40, SCC32, SCC35, and SCC37 on day 10 after inoculation. Four areas from each tumor and three tumors from each SCC were examined. Scale bars, 20 μm. DAPI staining is not shown. B, Representative CODEX multiplexed tissue images for CD3 (green), CD8 (red), TIM3 (magenta), and LAG3 (cyan blue) in tumors derived from SCC31, SCC40, SCC32, SCC35, and SCC37 on day 6 after inoculation. Five areas from each tumor and three tumors from each SCC were examined. Scale bars, 20 μm. C, Quantification of the percentage of F4/80 + CD204 + CD206 + cells described in A . Data are mean ± SEM. The rejected and nonrejected groups were compared using the Wilcoxon rank-sum test; W = 456; P = 2.55 × 10 −8 . D, Quantification of the percentage of CD3 + CD8 + cells described in B . Data are mean ± SEM. P = 0.00055; t = −5.27; degrees of freedom (df) = 9, unpaired t test. E, Quantification of the percentage of TIM3 + LAG3 + /CD3 + CD8 + cells described in B . Data are mean ± SEM. P = 0.02522; t = 2.6798; df = 9, unpaired t test. F, Boxplot quantifying the percentage of total cells for each CD8 cluster identified in CyTOF analysis. G, Boxplot quantifying the percentage of total cells for each macrophage cluster identified in CyTOF analysis. H, CFSE-based T cell proliferation assay after co-culture for 48 hours with nonrejected SCC35 tumor-derived macrophages isolated 10 days after inoculation. Representative histogram plot and quantification of CFSE intensity ( n = 5). CD8 + /CD4 + T cells isolated from healthy mouse spleens cultured without macrophages served as the controls. CD8: Kruskal–Wallis χ 2 test, χ 2 = 12.5; P = 0.001930454. Pairwise comparison by the Wilcoxon test with Bonferroni correction is shown in the figure. *, P value < 0.05. CD4: Kruskal–Wallis χ 2 test, χ 2 = 12.5; P = 0.00193. Pairwise comparison by the Wilcoxon test with Bonferroni correction is shown in the figure. *, P value < 0.05. ELISA measurement of IFNγ in culture media 48 hours after co-culture of T cells with or without tumor-derived macrophages. n = 6. Data are mean ± SEM. Kruskal–Wallis χ 2 test, χ 2 = 15.726; P = 0.0003847. Pairwise comparison by the Wilcoxon test with Bonferroni correction is shown. ***, P value < 0.01. MFI, mean fluorescence intensity.

Article Snippet: The samples were stained with TIM3 (Standard BioTools, Cat. #3162029, RRID: AB_2687841), CD8 (Standard BioTools, Cat. #3153012B, RRID: AB_2885019), Cx3cr1 (Standard BioTools, Cat. #92J020155, RRID: AB_3665151), CD62L (Standard BioTools, Cat. #3160008, RRID: AB_2687840), TCRβ (Fluidigm, Cat. #3143010B, RRID: AB_3665159), CD3 (Standard BioTools, Cat. #3152004, RRID: AB_2687836), CTLA-4 (Fluidigm, Cat. #3154008B, RRID: AB_3665152), PD-1 (Standard BioTools, Cat. #3159024, RRID: AB_2687839), CD44 (Standard BioTools, Cat. #3171003B, RRID: AB_2895121), GrnzB (Standard BioTools, Cat #3173006B, RRID: AB_2811095), Lag3 (Standard BioTools, Cat. #3174019B, RRID: AB_3665154), ICOS (Fluidigm, Cat. #3176014B, RRID: AB_3665155), CD4 (Standard BioTools, Cat. #3145002B, RRID: AB_2687832), CCR7 (Standard BioTools, Cat. #92J015163, RRID: AB_3665153), CD25 (Standard BioTools, Cat. #3151007B, RRID: AB_2827880), Foxp3 (Standard BioTools, Cat. #3165024, RRID: AB_2687843), MHC-II (Standard BioTools, Cat. #3209006B, RRID: AB_2885025), iNOS (Standard BioTools, Cat. #3161011B, RRID: AB_2922920), CD80 (Fluidigm, Cat. #92J023158, RRID: AB_3665158), F480 (Standard BioTools, Cat. #3146008B, RRID: AB_2895117), CD11b (Standard BioTools, Cat. #3148003B, RRID: AB_2814738), Ly6C (Standard BioTools, Cat. #3150010B, RRID: AB_2895118), CD206 (Standard BioTools, Cat. #3169021B, RRID: AB_2832249), CD86 (Standard BioTools, Cat. #3172016B, RRID: AB_2922923), and Ki67 (Standard BioTools, Cat. #3168007B, RRID: AB_2800467).

Techniques: Imaging, Multiplex Assay, Derivative Assay, Staining, Proliferation Assay, Co-Culture Assay, Isolation, Cell Culture, Comparison, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: Cancer Discovery

Article Title: Temporal Genomic Analysis of Homogeneous Tumor Models Reveals Key Regulators of Immune Evasion in Melanoma

doi: 10.1158/2159-8290.CD-23-1422

Figure Lengend Snippet: The decreased growth of Mif KO tumors is mediated by CD8 + T cells characterized by less exhaustion and high proliferation. A, Representative Opal Multiplex IHC stains for F4/80 (red), CD204 (yellow), and CD206 (green) in tumors derived from Mif WT or KO SCC35 on day 10 after inoculation. Four areas from each tumor and three tumors from each SCC were examined. Scale bars, 20 μm. DAPI staining is not shown. B, Quantification of the percentage of F4/80 + CD204 + CD206 + cells described in A . Data are mean ± SEM. P = 0.0012; t = −10.411; degrees of freedom (df) = 3.28, unpaired t test. C, Representative CODEX multiplexed tissue images for CD3 (green), CD8 (red), TIM3 (magenta), and LAG3 (cyan blue) in tumors derived from Mif WT or KO SCC35 on day 10 after inoculation. Five areas from each tumor and three and four tumors from Mif KO and Mif WT, respectively, were examined. Scale bars, 20 μm. D, Quantification of the percentage of CD3 + CD8 + T cells described in C . Data are mean ± SEM. P = 0.05732; t = 3.8252; df = 4, unpaired t test. E, Quantification of the percentage of TIM3 + LAG3 + /CD3 + CD8 + T cells described in C . Data are mean ± SEM. P = 0.004664; t = −6.0229; df = 3.759, unpaired t test. F, Growth curve of tumors derived from Mif KO clones (left: SCC37 KO clone 21; right: SCC35 KO clone 24) in immunocompetent mice, treated with anti-CD8 antibody or IgG control. n = 8–9. Pairwise comparisons using the Wilcoxon rank-sum test. P = 2.2 × 10 −5 and 0.00058, respectively.

Article Snippet: The samples were stained with TIM3 (Standard BioTools, Cat. #3162029, RRID: AB_2687841), CD8 (Standard BioTools, Cat. #3153012B, RRID: AB_2885019), Cx3cr1 (Standard BioTools, Cat. #92J020155, RRID: AB_3665151), CD62L (Standard BioTools, Cat. #3160008, RRID: AB_2687840), TCRβ (Fluidigm, Cat. #3143010B, RRID: AB_3665159), CD3 (Standard BioTools, Cat. #3152004, RRID: AB_2687836), CTLA-4 (Fluidigm, Cat. #3154008B, RRID: AB_3665152), PD-1 (Standard BioTools, Cat. #3159024, RRID: AB_2687839), CD44 (Standard BioTools, Cat. #3171003B, RRID: AB_2895121), GrnzB (Standard BioTools, Cat #3173006B, RRID: AB_2811095), Lag3 (Standard BioTools, Cat. #3174019B, RRID: AB_3665154), ICOS (Fluidigm, Cat. #3176014B, RRID: AB_3665155), CD4 (Standard BioTools, Cat. #3145002B, RRID: AB_2687832), CCR7 (Standard BioTools, Cat. #92J015163, RRID: AB_3665153), CD25 (Standard BioTools, Cat. #3151007B, RRID: AB_2827880), Foxp3 (Standard BioTools, Cat. #3165024, RRID: AB_2687843), MHC-II (Standard BioTools, Cat. #3209006B, RRID: AB_2885025), iNOS (Standard BioTools, Cat. #3161011B, RRID: AB_2922920), CD80 (Fluidigm, Cat. #92J023158, RRID: AB_3665158), F480 (Standard BioTools, Cat. #3146008B, RRID: AB_2895117), CD11b (Standard BioTools, Cat. #3148003B, RRID: AB_2814738), Ly6C (Standard BioTools, Cat. #3150010B, RRID: AB_2895118), CD206 (Standard BioTools, Cat. #3169021B, RRID: AB_2832249), CD86 (Standard BioTools, Cat. #3172016B, RRID: AB_2922923), and Ki67 (Standard BioTools, Cat. #3168007B, RRID: AB_2800467).

Techniques: Multiplex Assay, Derivative Assay, Staining, Clone Assay, Control