depletion antibodies anti mouse cd8α  (Bio X Cell)

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: Tumour-infiltrating immune cells isolated from the CD-fed, HFD-fed, or HFD + B. breve -treated mice were assessed by multicolour flow cytometry on Day 14 after tumour cell inoculation ( n = 8 mice/group). A , F NK cells, NKT cells, T cells, Tc cells, and Th cells among total single live cells in Hepa1-6 tumours. B , G Proportions of Tc cells and Th cells among the total population of T cells. C , H DCs, TAMs and their subsets, and MDSCs of total single live cells in Hepa1-6 tumours. D , I Proportions of proinflammatory (M1) TAMs and alternatively activated (M2) TAMs among the total population of TAMs. E , J Representative images of H&E-stained tumour tissues are shown in the left panel. Immunohistochemical staining of the tumour sections revealed that the main tumour-infiltrating immune cells were recognized by detecting the expression of corresponding surface markers, such as CD3, CD4, CD8, F4/80, CD86 and CD206 (left panel; scale bar = 50 μm). The quantitative analysis was performed with ImageJ, and the number of positive cells is presented in column plots (right panel). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns: not significant. CD control diet, HFD high-fat diet, B. breve Bifidobacterium breve , NK natural killer, Tc cytotoxic CD8 + T cell, Th helper CD4 + T cell, DCs dendritic cells, TAMs tumour-associated macrophages, MDSCs myeloid-derived suppressor cells.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: Isolation, Flow Cytometry, Staining, Immunohistochemical staining, Expressing, Control, Derivative Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: A Schematic of the experimental timeline in ob/ob+PBS-treated and ob/ob+ B. breve -treated mice ( n = 5 mice/group). B Body weight curves and comparison of body weights at the end point. C Wet weights of eWAT and liver and the liver/body weight ratio at the end of the experiment. D , F In vivo bioluminescence imaging at 9 days after the tumour cell injection ( D ) and the total radiant efficiency of the abdominal region ( F ). E , G Macroscopic representation (scale bar = 1 cm; the white arrow indicates the tumour region) and volume of the Hepa1-6-luc tumours. H NK cells, T cells, Tc cells, and Th cells among total single live cells in Hepa1-6 tumours. I Proportions of Tc cells and Th cells among the total T-cell population. J DCs, TAMs and their subsets, and MDSCs among the total single live cells in Hepa1-6 tumours. K Proportions of M1 TAMs and M2 TAMs among the total population of TAMs. The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, and ns not significant. B. breve Bifidobacterium breve , eWAT epididymal white adipose tissue, NK natural killer, Tc cytotoxic CD8 + T cell, Th helper CD4 + T cell, DCs dendritic cells, TAMs tumour-associated macrophages, MDSCs myeloid-derived suppressor cells.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: Comparison, In Vivo, Imaging, Injection, Derivative Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: A – E Serum from CD-fed, HFD-fed, or HFD + B. breve -treated mice was analysed by LC‒MS/MS to determine the metabolite composition. A PCA plot of the serum metabolomic analysis. B Venn diagram showing the shared and distinct metabolites among the three groups. C Heatmap with the normalization of the relative abundance of differentially abundant serum metabolites based on pairwise comparisons. D Absolute quantification of TCA and TDCA levels in the serum of the mice from the three groups. E Correlation analysis between Bifidobacterium and TCA levels in DIO mice. F B. breve decomposed TCA under in vitro culture conditions ( n = 4/group). G – M TCA exacerbates tumour development and restricts CD8 + T-cell infiltration. G Schematic of the experimental timeline in PBS-treated and TCA-treated mice (PBS: n = 5 mice/group, TCA: n = 6 mice/group). H Macroscopic representation, tumour growth curve and mass of Hepa1-6 tumours. I NK cells, T cells, Tc cells, and Th cells among total single live cells in Hepa1-6 tumours. J Proportions of Tc cells and Th cells among the total population of T cells. K DCs, TAMs and their subsets, and MDSCs among the total single live cells in Hepa1-6 tumours. L Proportions of M1 TAMs and M2 TAMs among the total population of TAMs. M Representative images of H&E staining and immunohistochemical staining of the tumour sections. The main tumour-infiltrating immune cells were recognized by detecting the expression of corresponding surface markers (left panel; scale bar = 50 μm). The quantitative analysis was performed with ImageJ, and the number of positive cells is presented in column plots (right panel). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns not significant. TCA taurocholic acid, CD control diet, HFD high-fat diet, B. breve Bifidobacterium breve , LC‒MS/MS liquid chromatography‒mass spectrometry, TDCA taurodeoxycholic acid, NK natural killer, Tc cytotoxic CD8 + T cell, Th helper CD4 + T cell, DCs dendritic cells, TAMs tumour-associated macrophages, MDSCs myeloid-derived suppressor cells.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: Quantitative Proteomics, In Vitro, Staining, Immunohistochemical staining, Expressing, Control, Derivative Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: A AAA-10 suppressed the BSH activity of B. breve in vitro. B Schematic of the experimental timeline of HFD + B. breve -treated and HFD + B. breve + AAA-10-treated mice ( n = 6 mice/group). C Macroscopic representation, growth curve and mass of Hepa1-6 tumours from AAA-10-treated and control mice. D Body weight curves and comparison of body weights at the 16th week. E Wet weights of eWAT and liver at the end of the experiment. F Representative images of livers stained with H&E (scale bar = 100 µm) and the lipid area, calculated as % AOI in liver cross-sections stained with H&E. G NK cells, NKT cells, T cells, Tc cells, and Th cells among total single live cells in Hepa1-6 tumours. H Proportions of Tc cells and Th cells among the total population of T cells. I DCs, TAMs and their subsets, and MDSCs among the total single live cells in Hepa1-6 tumours. J Proportions of M1 TAMs and M2 TAMs among the total population of TAMs. K Representative images of H&E staining and immunohistochemical staining of the tumour sections. The main tumour-infiltrating immune cells were identified by detecting the expression of corresponding surface markers (left panel; scale bar = 50 μm, Foxp3; scale bar = 100 μm). The quantitative analysis was performed with ImageJ, and the number of positive cells is presented in column plots (right panel). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, and ns not significant. BSH bile salt hydrolase, HFD high-fat diet, B. breve Bifidobacterium breve , eWAT epididymal white adipose tissue, AOI area of interest, H&E haematoxylin and eosin, NK natural killer, Tc cytotoxic CD8 + T cell, Th helper CD4 + T cell, DCs dendritic cells, TAMs tumour-associated macrophages, MDSCs myeloid-derived suppressor cells.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: Activity Assay, In Vitro, Control, Comparison, Staining, Immunohistochemical staining, Expressing, Derivative Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: A Schematic of the CD8 + T-cell depletion experimental timeline ( n = 6 mice/group). B Macroscopic representation, growth curve and mass of the Hepa1-6 tumours. C Tumour-infiltrating lymphocytes were analysed using flow cytometry ( n = 4‒6/group). D , E MACS-sorted CD8 + T cells isolated from naïve WT mice were activated and expanded with IL-2 and α-CD3/CD28 antibodies. The cells were treated with different doses of TCA for 24 h. D Apoptosis of CD8 + T cells treated with TCA in vitro. E Flow cytometry was used to evaluate proliferation and the expression of cytotoxic cytokines and chemokine receptors. F Expression of the mRNAs encoding cytotoxic cytokines and chemokine receptors in CD8 + T cells cultured in vitro was normalized to that of b-actin. The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns not significant. TCA taurocholic acid, CD control diet, WT wild type, MACS magnetic-activated cell sorting.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: Flow Cytometry, Isolation, In Vitro, Expressing, Cell Culture, Control, FACS

Journal: NPJ Biofilms and Microbiomes

Article Title: Deconjugating taurocholic acid with Bifidobacterium to mitigate obesity-driven cancer progression by restoring CD8 + T-cell infiltration

doi: 10.1038/s41522-025-00809-4

Figure Lengend Snippet: A Volcano plot comparing the mRNA profiles analysed with RNA-seq of CD8 + T cells isolated from the Hepa1-6 tumours of the TCA-treated and control mice. B Heatmaps illustrating the relative expression of signature genes in CD8 + T cells derived from the TCA-treated and control mice. C KEGG pathway enrichment analysis and D Analysis of GO- BP (biological process) terms of downregulated genes in the TCA-treated CD8 + T cells. The top 20 identified pathways are presented. E CD8 + T cells isolated from naïve WT mice and treated with different doses of TCA were analysed for the phosphorylation of ERK, P38, JNK, and P65 by immunoblotting. Column plots depict the relative band intensity ratios of phosphorylated/total proteins, and the relative expression was normalized to that of RPS-18. F – H Effects of treatment with TCA with or without C16-PAF on the phosphorylation of ERK ( F ), proliferation, and expression of cytotoxic cytokines and chemokine receptors at the transcript and protein levels in CD8 + T cells, as determined by flow cytometry ( G ) and qPCR ( H ). The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns not significant. TCA taurocholic acid, KEGG Kyoto Encyclopedia of Genes and Genomes, GO-BP Gene Ontology biological process terms, WT wild type, q-PCR quantitative real-time PCR.

Article Snippet: For the depletion of CD8 + T cells, we injected mice with an anti-CD8 depletion antibody (Bio X Cell, BE0061, 2.43) or an isotype control antibody (Bio X Cell, BE0090, LTF-2) at a dose of 250 μg per mouse intraperitoneally starting on day -3 of tumour cell inoculation and every 3 days thereafter until the end of the study.

Techniques: RNA Sequencing, Isolation, Control, Expressing, Derivative Assay, Phospho-proteomics, Western Blot, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Redox biology

Article Title: Radiation-induced upregulation of itaconate in macrophages promotes the radioresistance of non-small cell lung cancer by stabilizing NRF2 protein and suppressing immune response.

doi: 10.1016/j.redox.2025.103711

Figure Lengend Snippet: Fig. 5. Knockout of Acod1 in macrophages further enhance the activation of tumor immune microenvironment by radiotherapy (A) Gating strategy for detection of NK, CD4+ and CD8+ T cells by flow cytometry. (B–G) Flow cytometry analysis of CD45+ immune cells (B), CD3+ (C), CD4+ (D), CD8+ (E) T cells, the ratio of CD4+/CD8+ T cells (F), and NK cells (G) in the TME of LLC subcutaneous tumor model in mice treated with radiation (8 Gy x 3) in Acod1f/f Lyz2cre− (HO−) and Acod1f/f Lyz2cre+ (HO+) mice (n = 6). (H) Representative immunofluorescence staining of CD8 (green) in the TME at the end of experiment in Acod1f/f Lyz2cre−(HO−) and Acod1f/f Lyz2cre+ (HO+) mice. (I–J) Flow cytometry analysis of CD107a+ (I), and IFN-γ+ CD8+ T cells (J) in the TME of LLC subcutaneous tumor model described above (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001; ns, not statistically significant.

Article Snippet: CD8+ T cell depletion Anti-mouse CD8 monoclonal antibody (clone 2.43) (#A2102, Selleck, China) was intraperitoneally injected at a dose of 200 μg per mouse, and 150 μg per mouse was injected 3 days later.

Techniques: Knock-Out, Activation Assay, Flow Cytometry, Immunofluorescence, Staining

Journal: Redox biology

Article Title: Radiation-induced upregulation of itaconate in macrophages promotes the radioresistance of non-small cell lung cancer by stabilizing NRF2 protein and suppressing immune response.

doi: 10.1016/j.redox.2025.103711

Figure Lengend Snippet: Fig. 6. The antitumor effect of Acod1 knockout in macrophages combined with radiotherapy partially depends on CD8+ T cells (A) CD8+ T cells clearance efficiency in mouse spleen detected by flow cytometry (n = 3). (B) CD8+ T cells clearance efficiency in mouse peripheral blood detected by flow cytometry (n = 3). (C) Tumor growth curves of LLC subcutaneous tumor in Acod1f/f Lyz2cre−(HO−) and Acod1f/f Lyz2cre+ (HO+) mice treated with 8 Gy x 3 radiotherapy and IgG or neutralizing CD8 antibody (α-CD8) (n = 6). (D–E) Tumor weight (D) and tumor images (E) on day 16 after treatment in different groups (n = 6). (F) The schematic diagram depicting that macrophages up-regulate the expression of Acod1 through activating NF-κB pathway after radiotherapy, thereby regulating the radiosensitivity of tumor cells and immune cell infiltration. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not statistically significant.

Article Snippet: CD8+ T cell depletion Anti-mouse CD8 monoclonal antibody (clone 2.43) (#A2102, Selleck, China) was intraperitoneally injected at a dose of 200 μg per mouse, and 150 μg per mouse was injected 3 days later.

Techniques: Knock-Out, Flow Cytometry, Expressing