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cd8a ly 2 microbeads  (Miltenyi Biotec)


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    Miltenyi Biotec cd8a ly 2 microbeads
    <t>CD8</t> + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.
    Cd8a Ly 2 Microbeads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection"

    Article Title: CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection

    Journal: iScience

    doi: 10.1016/j.isci.2026.116611

    CD8 + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.
    Figure Legend Snippet: CD8 + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.

    Techniques Used: Infection, Single Cell, Flow Cytometry

    CD8 + cytotoxic T cells contribute as central mediators within the CCL signaling network (A) Violin plots show the distribution and density of CCL5 (left) and CCR5 (right) expression across T and NK cell subsets, as well as macrophages, in the colon of naive and chronically infected mice. Wider sections of the violin indicate a higher proportion of cells expressing CCL5 or CCR5 at that level. Data were generated from CellChat-guided analysis of scRNA-seq profiles. (B) Representative flow cytometry plots show CD8 + CCR5 + T cells in the colon of naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD3 + cells. (C) Representative flow cytometry plots show F4/80 + CCR5 + T cells in naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD45 + cells. (D) Violin plot shows the frequency of CD8 + CCR5 + T cells in naive and chronically infected mice. Data are representative of two independent experiments ( n = 4 mice per group). (E) mRNA quantification of Ccl5 and Ccr5 by qRT-PCR in naive and chronically infected mice. The plot shows log 2 fold changes (FC) in gene expression determined by the ΔC t method with normalization to GAPDH. The horizontal line indicates the median value for each group, with individual data points plotted. CD8 + T cells were purified from the lamina propria of naive and chronically infected mice using magnetic bead–based separation prior to RNA extraction. Data are from three independent experiments. (F) Violin plot shows the frequency of F4/80 + CCR5 + T cells in the colon of naive and chronically infected mice. Data are representative of two independent experiments ( n = 3 mice per group). In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗: p < 0.01; ns: not significant.
    Figure Legend Snippet: CD8 + cytotoxic T cells contribute as central mediators within the CCL signaling network (A) Violin plots show the distribution and density of CCL5 (left) and CCR5 (right) expression across T and NK cell subsets, as well as macrophages, in the colon of naive and chronically infected mice. Wider sections of the violin indicate a higher proportion of cells expressing CCL5 or CCR5 at that level. Data were generated from CellChat-guided analysis of scRNA-seq profiles. (B) Representative flow cytometry plots show CD8 + CCR5 + T cells in the colon of naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD3 + cells. (C) Representative flow cytometry plots show F4/80 + CCR5 + T cells in naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD45 + cells. (D) Violin plot shows the frequency of CD8 + CCR5 + T cells in naive and chronically infected mice. Data are representative of two independent experiments ( n = 4 mice per group). (E) mRNA quantification of Ccl5 and Ccr5 by qRT-PCR in naive and chronically infected mice. The plot shows log 2 fold changes (FC) in gene expression determined by the ΔC t method with normalization to GAPDH. The horizontal line indicates the median value for each group, with individual data points plotted. CD8 + T cells were purified from the lamina propria of naive and chronically infected mice using magnetic bead–based separation prior to RNA extraction. Data are from three independent experiments. (F) Violin plot shows the frequency of F4/80 + CCR5 + T cells in the colon of naive and chronically infected mice. Data are representative of two independent experiments ( n = 3 mice per group). In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗: p < 0.01; ns: not significant.

    Techniques Used: Expressing, Infection, Generated, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Purification, RNA Extraction

    CD8 + cytotoxic T cell–macrophage interactions likely shape colonic immune homeostasis during chronic T. cruzi infection (A) Volcano plot showing differentially expressed genes (DEGs) in lamina propria CD8 + cytotoxic T cells from infected mice compared with naive controls. Red dots depict upregulated genes, blue dots indicate downregulated genes, and gray dots represent genes with a non-significant change in expression. Genes with log 2 fold change ≥1.5, and p < 0.05 were considered significant. (B) Top five differentially regulated canonical pathways in lamina propria CD8 + cytotoxic T cells from infected mice, as predicted by ingenuity pathway analysis (IPA). Bars represent −log ( p value); red indicates pathway activation and blue indicates pathway inhibition. (C) Top 10 upregulated genes associated with the IL-4/IL-13 signaling pathway (left) and the pathogen-induced cytokine storm (PICS) pathway (right) in CD8 + cytotoxic T cells. (D) Volcano plot of DEGs in lamina propria macrophages from infected mice compared with naive controls, displayed as in (A). (E) Top five differentially regulated canonical pathways in lamina propria macrophages from infected mice, as predicted by IPA, displayed as in (B). (F) Top 10 upregulated genes associated with the S100 protein family signaling pathway (left) and the neutrophil degranulation pathway (right) in macrophages. In (C and F), bars indicate log 2 fold change between infected and naive mice.
    Figure Legend Snippet: CD8 + cytotoxic T cell–macrophage interactions likely shape colonic immune homeostasis during chronic T. cruzi infection (A) Volcano plot showing differentially expressed genes (DEGs) in lamina propria CD8 + cytotoxic T cells from infected mice compared with naive controls. Red dots depict upregulated genes, blue dots indicate downregulated genes, and gray dots represent genes with a non-significant change in expression. Genes with log 2 fold change ≥1.5, and p < 0.05 were considered significant. (B) Top five differentially regulated canonical pathways in lamina propria CD8 + cytotoxic T cells from infected mice, as predicted by ingenuity pathway analysis (IPA). Bars represent −log ( p value); red indicates pathway activation and blue indicates pathway inhibition. (C) Top 10 upregulated genes associated with the IL-4/IL-13 signaling pathway (left) and the pathogen-induced cytokine storm (PICS) pathway (right) in CD8 + cytotoxic T cells. (D) Volcano plot of DEGs in lamina propria macrophages from infected mice compared with naive controls, displayed as in (A). (E) Top five differentially regulated canonical pathways in lamina propria macrophages from infected mice, as predicted by IPA, displayed as in (B). (F) Top 10 upregulated genes associated with the S100 protein family signaling pathway (left) and the neutrophil degranulation pathway (right) in macrophages. In (C and F), bars indicate log 2 fold change between infected and naive mice.

    Techniques Used: Infection, Expressing, Activation Assay, Inhibition

    Proposed model of the immune response mechanisms to Trypanosoma cruzi in the mouse colon, integrating our findings with prior studies ① Infected smooth muscle cells release trypomastigotes into the surrounding colonic tissue. ② Trypomastigotes are phagocytosed by macrophages. ③ After 18–24 hs post-internalization, amastigotes are released into the cytoplasm, where they replicate and synthesize proteins that are presented on the macrophage surface. ④ CD8 + and CD4 + T cells recognize T. cruzi peptides presented by MHC complexes. ⑤ Upon activation, CD8 + T cells secrete granzyme A and multiple cytokines. ⑥ Granzyme A -containing granules may enter parasite-infected smooth muscle cells through pores formed by perforin-1 (PRF1; secreted by NK cells), contributing to cell death. ⑦ Tumor necrosis factor (TNF) may promote macrophage polarization to an M1 phenotype by inducing the expression of inducible nitric oxide synthase (NOS2). This enzyme subsequently oxidizes arginine to produce nitric oxide, which may contribute to parasite killing. ⑧ Activated CD4 + T cells can differentiate into Th and Tfh cells that produce Interleukin-4 (IL-4) and Interleukin-13 (IL-13) [⑨], which in turn induce M2 macrophage polarization. ⑩ C-C Motif Ligand 5 (CCL5) could further support M2 macrophage polarization via arginase I (ARG1) upregulation, promoting polyamines associated with parasite multiplication and persistence. ⑪ Amastigotes differentiate into trypomastigotes, completing the cycle by infecting naive smooth muscle cells.
    Figure Legend Snippet: Proposed model of the immune response mechanisms to Trypanosoma cruzi in the mouse colon, integrating our findings with prior studies ① Infected smooth muscle cells release trypomastigotes into the surrounding colonic tissue. ② Trypomastigotes are phagocytosed by macrophages. ③ After 18–24 hs post-internalization, amastigotes are released into the cytoplasm, where they replicate and synthesize proteins that are presented on the macrophage surface. ④ CD8 + and CD4 + T cells recognize T. cruzi peptides presented by MHC complexes. ⑤ Upon activation, CD8 + T cells secrete granzyme A and multiple cytokines. ⑥ Granzyme A -containing granules may enter parasite-infected smooth muscle cells through pores formed by perforin-1 (PRF1; secreted by NK cells), contributing to cell death. ⑦ Tumor necrosis factor (TNF) may promote macrophage polarization to an M1 phenotype by inducing the expression of inducible nitric oxide synthase (NOS2). This enzyme subsequently oxidizes arginine to produce nitric oxide, which may contribute to parasite killing. ⑧ Activated CD4 + T cells can differentiate into Th and Tfh cells that produce Interleukin-4 (IL-4) and Interleukin-13 (IL-13) [⑨], which in turn induce M2 macrophage polarization. ⑩ C-C Motif Ligand 5 (CCL5) could further support M2 macrophage polarization via arginase I (ARG1) upregulation, promoting polyamines associated with parasite multiplication and persistence. ⑪ Amastigotes differentiate into trypomastigotes, completing the cycle by infecting naive smooth muscle cells.

    Techniques Used: Infection, Activation Assay, Expressing

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    Article Title: Recent thymic emigrants preferentially undergo memory inflation after persistent infection.
    Article Snippet: .. CD8 + T cells were bead enriched from the spleens by positive selection with CD8a microbeads (Miltenyi). ..

    Article Title: CCL24 recruits CCR3 + TAMs to promote immunosuppression via YAP1 activation and serves as a therapeutic target for Gracillin in colorectal cancer
    Article Snippet: .. Thymus from C57BL/6J mice (6 weeks old, Anburui Biotechnologies, Fujian, China) was mechanically dissociated to prepare a single-cell suspension, and CD8 + T cells were purified by positive selection using CD8a microbeads (Miltenyi Biotec). .. The sorted CD8 + T cells were activated with CD3/CD28 agonist antibodies (2 μg/mL, BioLegend) for 48 h. Cell function was assessed by intracellular cytokine staining and flow cytometry to evaluate TNFA and GZMB expression.

    FACS:

    Article Title: Histone lactylation-driven feedback loop modulates cholesterol-linked immunosuppression in pancreatic cancer
    Article Snippet: .. Using CD8a microbeads and magnetic cell sorting (Miltenyi Biotec, Germany), CD8+ T cells were extracted from the spleens of OT-1 mice in preparation for OT-1 CD8+ T Cell adoptive transfer. ..

    Article Title: Histone lactylation-driven feedback loop modulates cholesterol-linked immunosuppression in pancreatic cancer
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    Adoptive Transfer Assay:

    Article Title: Histone lactylation-driven feedback loop modulates cholesterol-linked immunosuppression in pancreatic cancer
    Article Snippet: .. Using CD8a microbeads and magnetic cell sorting (Miltenyi Biotec, Germany), CD8+ T cells were extracted from the spleens of OT-1 mice in preparation for OT-1 CD8+ T Cell adoptive transfer. ..

    Single Cell:

    Article Title: CCL24 recruits CCR3 + TAMs to promote immunosuppression via YAP1 activation and serves as a therapeutic target for Gracillin in colorectal cancer
    Article Snippet: .. Thymus from C57BL/6J mice (6 weeks old, Anburui Biotechnologies, Fujian, China) was mechanically dissociated to prepare a single-cell suspension, and CD8 + T cells were purified by positive selection using CD8a microbeads (Miltenyi Biotec). .. The sorted CD8 + T cells were activated with CD3/CD28 agonist antibodies (2 μg/mL, BioLegend) for 48 h. Cell function was assessed by intracellular cytokine staining and flow cytometry to evaluate TNFA and GZMB expression.

    Article Title: LDLR + Monocytic Myeloid-derived Suppressor Cells Attenuate Allograft Rejection in Liver Transplantation
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    Suspension:

    Article Title: CCL24 recruits CCR3 + TAMs to promote immunosuppression via YAP1 activation and serves as a therapeutic target for Gracillin in colorectal cancer
    Article Snippet: .. Thymus from C57BL/6J mice (6 weeks old, Anburui Biotechnologies, Fujian, China) was mechanically dissociated to prepare a single-cell suspension, and CD8 + T cells were purified by positive selection using CD8a microbeads (Miltenyi Biotec). .. The sorted CD8 + T cells were activated with CD3/CD28 agonist antibodies (2 μg/mL, BioLegend) for 48 h. Cell function was assessed by intracellular cytokine staining and flow cytometry to evaluate TNFA and GZMB expression.

    Purification:

    Article Title: CCL24 recruits CCR3 + TAMs to promote immunosuppression via YAP1 activation and serves as a therapeutic target for Gracillin in colorectal cancer
    Article Snippet: .. Thymus from C57BL/6J mice (6 weeks old, Anburui Biotechnologies, Fujian, China) was mechanically dissociated to prepare a single-cell suspension, and CD8 + T cells were purified by positive selection using CD8a microbeads (Miltenyi Biotec). .. The sorted CD8 + T cells were activated with CD3/CD28 agonist antibodies (2 μg/mL, BioLegend) for 48 h. Cell function was assessed by intracellular cytokine staining and flow cytometry to evaluate TNFA and GZMB expression.

    Cell Isolation:

    Article Title: Histone lactylation-driven feedback loop modulates cholesterol-linked immunosuppression in pancreatic cancer
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    Article Title: Inhibition of Galectin-3 Expression Could Preserve the Immunoregulatory Function of Mesenchymal Stem Cells During Cell Passaging
    Article Snippet: .. CD4+/CD8+ T cell isolation or depletion was performed using CD4 and CD8a MicroBeads and the AutoMACS system (Miltenyi Biotec, Germany) according to the manufacturer’s protocol. .. Briefly, 10 μL of CD4 MicroBeads and 10 μL of CD8a MicroBeads were added per 1 × 107 cells, and the mixture was incubated for 15 minutes at 4 °C in the dark.

    Suppression Assay:

    Article Title: Histone lactylation-driven feedback loop modulates cholesterol-linked immunosuppression in pancreatic cancer
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    <t>CD8</t> + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.
    Pbs, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd8a microbeads kit  (Miltenyi Biotec)
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    Single-cell RNA sequencing (scRNA-seq) analysis of the tumor microenvironment (TME) in TNBC patients. (A) UMAP visualization of major cell populations within TNBC tumors. (B) Comparative analysis of immune cell frequencies in the TME of TNBC patients with and without chemotherapy. (C) UMAP visualization of macrophages within TNBC tumors. (D) Changes in macrophage subtype frequencies in the TME of TNBC patients with and without chemotherapy. (E) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by CD163 high <t>CD8A</t> low and CD163 low CD8A high expression ( P = 0.016). (F) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by ARG1 high CD8A low and ARG1 low CD8A high expression ( P = 0.049). (G) Representative immunohistochemical (IHC) staining of CD8 and CD163 in tumor tissue microarrays from TNBC patients. Scale bar = 100 μm. (H) Correlation analysis of CD8 and CD163 expression in chemotherapy-treated versus untreated TNBC patients (Patient 18,383: untreated; Patient 15,840: treated). (I) Two major clinical chemotherapies in the 4T1 murine triple negative breast cancer model time schedule. (J) Tumor volume analysis (K), Tumor weight (L), CD8/CD45, and (M) M1/M2 in TME. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.
    Cd8a Microbeads Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cd8a (ly-2) microbeads, mouse  (Miltenyi Biotec)
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    Single-cell RNA sequencing (scRNA-seq) analysis of the tumor microenvironment (TME) in TNBC patients. (A) UMAP visualization of major cell populations within TNBC tumors. (B) Comparative analysis of immune cell frequencies in the TME of TNBC patients with and without chemotherapy. (C) UMAP visualization of macrophages within TNBC tumors. (D) Changes in macrophage subtype frequencies in the TME of TNBC patients with and without chemotherapy. (E) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by CD163 high <t>CD8A</t> low and CD163 low CD8A high expression ( P = 0.016). (F) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by ARG1 high CD8A low and ARG1 low CD8A high expression ( P = 0.049). (G) Representative immunohistochemical (IHC) staining of CD8 and CD163 in tumor tissue microarrays from TNBC patients. Scale bar = 100 μm. (H) Correlation analysis of CD8 and CD163 expression in chemotherapy-treated versus untreated TNBC patients (Patient 18,383: untreated; Patient 15,840: treated). (I) Two major clinical chemotherapies in the 4T1 murine triple negative breast cancer model time schedule. (J) Tumor volume analysis (K), Tumor weight (L), CD8/CD45, and (M) M1/M2 in TME. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.
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    mouse cd8 ly 2 microbeads kit  (Miltenyi Biotec)
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    Single-cell RNA sequencing (scRNA-seq) analysis of the tumor microenvironment (TME) in TNBC patients. (A) UMAP visualization of major cell populations within TNBC tumors. (B) Comparative analysis of immune cell frequencies in the TME of TNBC patients with and without chemotherapy. (C) UMAP visualization of macrophages within TNBC tumors. (D) Changes in macrophage subtype frequencies in the TME of TNBC patients with and without chemotherapy. (E) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by CD163 high <t>CD8A</t> low and CD163 low CD8A high expression ( P = 0.016). (F) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by ARG1 high CD8A low and ARG1 low CD8A high expression ( P = 0.049). (G) Representative immunohistochemical (IHC) staining of CD8 and CD163 in tumor tissue microarrays from TNBC patients. Scale bar = 100 μm. (H) Correlation analysis of CD8 and CD163 expression in chemotherapy-treated versus untreated TNBC patients (Patient 18,383: untreated; Patient 15,840: treated). (I) Two major clinical chemotherapies in the 4T1 murine triple negative breast cancer model time schedule. (J) Tumor volume analysis (K), Tumor weight (L), CD8/CD45, and (M) M1/M2 in TME. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.
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    Image Search Results


    CD8 + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.

    Journal: iScience

    Article Title: CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection

    doi: 10.1016/j.isci.2026.116611

    Figure Lengend Snippet: CD8 + T cytotoxic cells are the predominant subset within the T and NK cell compartment in the colon of chronically infected mice (A) Uniform manifold approximation and projection (UMAP) plot showing nine T and NK cell subclusters. Each point represents a single cell. (B) Bar graph depicts the relative percentages of each cell subtype in naive and infected mice. (C) Representative flow cytometry plots of CD8 + T cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + cells. (D) Violin plot shows the percentage of CD8 + T cells in naive and infected mice. (E) Representative flow cytometry plots of T follicular helper cells in naive (left) and infected mice (right). Cells were gated on live, singlets, CD3 + CD4 + cells. (F) Violin plot shows the percentage of PD-1 + CXCR4 + T follicular helper cells in naive and chronically infected mice. Data are representative of two independent experiments with a total of 6 (in D) or 4 (in F) mice per group. In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗∗: p < 0.001; ns: not significant.

    Article Snippet: CD8a (Ly-2) Microbeads , Miltenyi Biotec , Cat-130-117-044.

    Techniques: Infection, Single Cell, Flow Cytometry

    CD8 + cytotoxic T cells contribute as central mediators within the CCL signaling network (A) Violin plots show the distribution and density of CCL5 (left) and CCR5 (right) expression across T and NK cell subsets, as well as macrophages, in the colon of naive and chronically infected mice. Wider sections of the violin indicate a higher proportion of cells expressing CCL5 or CCR5 at that level. Data were generated from CellChat-guided analysis of scRNA-seq profiles. (B) Representative flow cytometry plots show CD8 + CCR5 + T cells in the colon of naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD3 + cells. (C) Representative flow cytometry plots show F4/80 + CCR5 + T cells in naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD45 + cells. (D) Violin plot shows the frequency of CD8 + CCR5 + T cells in naive and chronically infected mice. Data are representative of two independent experiments ( n = 4 mice per group). (E) mRNA quantification of Ccl5 and Ccr5 by qRT-PCR in naive and chronically infected mice. The plot shows log 2 fold changes (FC) in gene expression determined by the ΔC t method with normalization to GAPDH. The horizontal line indicates the median value for each group, with individual data points plotted. CD8 + T cells were purified from the lamina propria of naive and chronically infected mice using magnetic bead–based separation prior to RNA extraction. Data are from three independent experiments. (F) Violin plot shows the frequency of F4/80 + CCR5 + T cells in the colon of naive and chronically infected mice. Data are representative of two independent experiments ( n = 3 mice per group). In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗: p < 0.01; ns: not significant.

    Journal: iScience

    Article Title: CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection

    doi: 10.1016/j.isci.2026.116611

    Figure Lengend Snippet: CD8 + cytotoxic T cells contribute as central mediators within the CCL signaling network (A) Violin plots show the distribution and density of CCL5 (left) and CCR5 (right) expression across T and NK cell subsets, as well as macrophages, in the colon of naive and chronically infected mice. Wider sections of the violin indicate a higher proportion of cells expressing CCL5 or CCR5 at that level. Data were generated from CellChat-guided analysis of scRNA-seq profiles. (B) Representative flow cytometry plots show CD8 + CCR5 + T cells in the colon of naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD3 + cells. (C) Representative flow cytometry plots show F4/80 + CCR5 + T cells in naive (upper panel) and chronically infected mice (bottom panel). Cells were gated on live, singlets, and CD45 + cells. (D) Violin plot shows the frequency of CD8 + CCR5 + T cells in naive and chronically infected mice. Data are representative of two independent experiments ( n = 4 mice per group). (E) mRNA quantification of Ccl5 and Ccr5 by qRT-PCR in naive and chronically infected mice. The plot shows log 2 fold changes (FC) in gene expression determined by the ΔC t method with normalization to GAPDH. The horizontal line indicates the median value for each group, with individual data points plotted. CD8 + T cells were purified from the lamina propria of naive and chronically infected mice using magnetic bead–based separation prior to RNA extraction. Data are from three independent experiments. (F) Violin plot shows the frequency of F4/80 + CCR5 + T cells in the colon of naive and chronically infected mice. Data are representative of two independent experiments ( n = 3 mice per group). In (D and F), violin plots show the distribution of values for each group. Individual points represent biological replicates. Large dashed horizontal lines indicate the median. Statistical significance was determined by an unpaired t test. ∗∗: p < 0.01; ns: not significant.

    Article Snippet: CD8a (Ly-2) Microbeads , Miltenyi Biotec , Cat-130-117-044.

    Techniques: Expressing, Infection, Generated, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Purification, RNA Extraction

    CD8 + cytotoxic T cell–macrophage interactions likely shape colonic immune homeostasis during chronic T. cruzi infection (A) Volcano plot showing differentially expressed genes (DEGs) in lamina propria CD8 + cytotoxic T cells from infected mice compared with naive controls. Red dots depict upregulated genes, blue dots indicate downregulated genes, and gray dots represent genes with a non-significant change in expression. Genes with log 2 fold change ≥1.5, and p < 0.05 were considered significant. (B) Top five differentially regulated canonical pathways in lamina propria CD8 + cytotoxic T cells from infected mice, as predicted by ingenuity pathway analysis (IPA). Bars represent −log ( p value); red indicates pathway activation and blue indicates pathway inhibition. (C) Top 10 upregulated genes associated with the IL-4/IL-13 signaling pathway (left) and the pathogen-induced cytokine storm (PICS) pathway (right) in CD8 + cytotoxic T cells. (D) Volcano plot of DEGs in lamina propria macrophages from infected mice compared with naive controls, displayed as in (A). (E) Top five differentially regulated canonical pathways in lamina propria macrophages from infected mice, as predicted by IPA, displayed as in (B). (F) Top 10 upregulated genes associated with the S100 protein family signaling pathway (left) and the neutrophil degranulation pathway (right) in macrophages. In (C and F), bars indicate log 2 fold change between infected and naive mice.

    Journal: iScience

    Article Title: CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection

    doi: 10.1016/j.isci.2026.116611

    Figure Lengend Snippet: CD8 + cytotoxic T cell–macrophage interactions likely shape colonic immune homeostasis during chronic T. cruzi infection (A) Volcano plot showing differentially expressed genes (DEGs) in lamina propria CD8 + cytotoxic T cells from infected mice compared with naive controls. Red dots depict upregulated genes, blue dots indicate downregulated genes, and gray dots represent genes with a non-significant change in expression. Genes with log 2 fold change ≥1.5, and p < 0.05 were considered significant. (B) Top five differentially regulated canonical pathways in lamina propria CD8 + cytotoxic T cells from infected mice, as predicted by ingenuity pathway analysis (IPA). Bars represent −log ( p value); red indicates pathway activation and blue indicates pathway inhibition. (C) Top 10 upregulated genes associated with the IL-4/IL-13 signaling pathway (left) and the pathogen-induced cytokine storm (PICS) pathway (right) in CD8 + cytotoxic T cells. (D) Volcano plot of DEGs in lamina propria macrophages from infected mice compared with naive controls, displayed as in (A). (E) Top five differentially regulated canonical pathways in lamina propria macrophages from infected mice, as predicted by IPA, displayed as in (B). (F) Top 10 upregulated genes associated with the S100 protein family signaling pathway (left) and the neutrophil degranulation pathway (right) in macrophages. In (C and F), bars indicate log 2 fold change between infected and naive mice.

    Article Snippet: CD8a (Ly-2) Microbeads , Miltenyi Biotec , Cat-130-117-044.

    Techniques: Infection, Expressing, Activation Assay, Inhibition

    Proposed model of the immune response mechanisms to Trypanosoma cruzi in the mouse colon, integrating our findings with prior studies ① Infected smooth muscle cells release trypomastigotes into the surrounding colonic tissue. ② Trypomastigotes are phagocytosed by macrophages. ③ After 18–24 hs post-internalization, amastigotes are released into the cytoplasm, where they replicate and synthesize proteins that are presented on the macrophage surface. ④ CD8 + and CD4 + T cells recognize T. cruzi peptides presented by MHC complexes. ⑤ Upon activation, CD8 + T cells secrete granzyme A and multiple cytokines. ⑥ Granzyme A -containing granules may enter parasite-infected smooth muscle cells through pores formed by perforin-1 (PRF1; secreted by NK cells), contributing to cell death. ⑦ Tumor necrosis factor (TNF) may promote macrophage polarization to an M1 phenotype by inducing the expression of inducible nitric oxide synthase (NOS2). This enzyme subsequently oxidizes arginine to produce nitric oxide, which may contribute to parasite killing. ⑧ Activated CD4 + T cells can differentiate into Th and Tfh cells that produce Interleukin-4 (IL-4) and Interleukin-13 (IL-13) [⑨], which in turn induce M2 macrophage polarization. ⑩ C-C Motif Ligand 5 (CCL5) could further support M2 macrophage polarization via arginase I (ARG1) upregulation, promoting polyamines associated with parasite multiplication and persistence. ⑪ Amastigotes differentiate into trypomastigotes, completing the cycle by infecting naive smooth muscle cells.

    Journal: iScience

    Article Title: CCL signaling drives T Cell–Macrophage crosstalk in the mouse colon during chronic Trypanosoma cruzi infection

    doi: 10.1016/j.isci.2026.116611

    Figure Lengend Snippet: Proposed model of the immune response mechanisms to Trypanosoma cruzi in the mouse colon, integrating our findings with prior studies ① Infected smooth muscle cells release trypomastigotes into the surrounding colonic tissue. ② Trypomastigotes are phagocytosed by macrophages. ③ After 18–24 hs post-internalization, amastigotes are released into the cytoplasm, where they replicate and synthesize proteins that are presented on the macrophage surface. ④ CD8 + and CD4 + T cells recognize T. cruzi peptides presented by MHC complexes. ⑤ Upon activation, CD8 + T cells secrete granzyme A and multiple cytokines. ⑥ Granzyme A -containing granules may enter parasite-infected smooth muscle cells through pores formed by perforin-1 (PRF1; secreted by NK cells), contributing to cell death. ⑦ Tumor necrosis factor (TNF) may promote macrophage polarization to an M1 phenotype by inducing the expression of inducible nitric oxide synthase (NOS2). This enzyme subsequently oxidizes arginine to produce nitric oxide, which may contribute to parasite killing. ⑧ Activated CD4 + T cells can differentiate into Th and Tfh cells that produce Interleukin-4 (IL-4) and Interleukin-13 (IL-13) [⑨], which in turn induce M2 macrophage polarization. ⑩ C-C Motif Ligand 5 (CCL5) could further support M2 macrophage polarization via arginase I (ARG1) upregulation, promoting polyamines associated with parasite multiplication and persistence. ⑪ Amastigotes differentiate into trypomastigotes, completing the cycle by infecting naive smooth muscle cells.

    Article Snippet: CD8a (Ly-2) Microbeads , Miltenyi Biotec , Cat-130-117-044.

    Techniques: Infection, Activation Assay, Expressing

    Single-cell RNA sequencing (scRNA-seq) analysis of the tumor microenvironment (TME) in TNBC patients. (A) UMAP visualization of major cell populations within TNBC tumors. (B) Comparative analysis of immune cell frequencies in the TME of TNBC patients with and without chemotherapy. (C) UMAP visualization of macrophages within TNBC tumors. (D) Changes in macrophage subtype frequencies in the TME of TNBC patients with and without chemotherapy. (E) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by CD163 high CD8A low and CD163 low CD8A high expression ( P = 0.016). (F) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by ARG1 high CD8A low and ARG1 low CD8A high expression ( P = 0.049). (G) Representative immunohistochemical (IHC) staining of CD8 and CD163 in tumor tissue microarrays from TNBC patients. Scale bar = 100 μm. (H) Correlation analysis of CD8 and CD163 expression in chemotherapy-treated versus untreated TNBC patients (Patient 18,383: untreated; Patient 15,840: treated). (I) Two major clinical chemotherapies in the 4T1 murine triple negative breast cancer model time schedule. (J) Tumor volume analysis (K), Tumor weight (L), CD8/CD45, and (M) M1/M2 in TME. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: Astragalus-derived nano-agonist potentiates chemotherapy by reducing tumor-suppressive macrophages

    doi: 10.1016/j.apsb.2026.03.051

    Figure Lengend Snippet: Single-cell RNA sequencing (scRNA-seq) analysis of the tumor microenvironment (TME) in TNBC patients. (A) UMAP visualization of major cell populations within TNBC tumors. (B) Comparative analysis of immune cell frequencies in the TME of TNBC patients with and without chemotherapy. (C) UMAP visualization of macrophages within TNBC tumors. (D) Changes in macrophage subtype frequencies in the TME of TNBC patients with and without chemotherapy. (E) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by CD163 high CD8A low and CD163 low CD8A high expression ( P = 0.016). (F) Kaplan–Meier survival analysis of TNBC patients from the TCGA database stratified by ARG1 high CD8A low and ARG1 low CD8A high expression ( P = 0.049). (G) Representative immunohistochemical (IHC) staining of CD8 and CD163 in tumor tissue microarrays from TNBC patients. Scale bar = 100 μm. (H) Correlation analysis of CD8 and CD163 expression in chemotherapy-treated versus untreated TNBC patients (Patient 18,383: untreated; Patient 15,840: treated). (I) Two major clinical chemotherapies in the 4T1 murine triple negative breast cancer model time schedule. (J) Tumor volume analysis (K), Tumor weight (L), CD8/CD45, and (M) M1/M2 in TME. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

    Article Snippet: CD8 + T lymphocytes were extracted from the spleen of BALB/c mice with the CD8a microbeads kit (Miltenyi, 130-117-044, Bergisch Gladbach, Germany).

    Techniques: Single Cell, RNA Sequencing, Expressing, Immunohistochemical staining, Immunohistochemistry

    Analysis of T lymphocyte phenotypes and macrophage subtypes in TME. (A) Proportion of CD45 cells in live cells ( n = 6 per group). (B) Ratio of macrophages to CD45 + T cells ( n = 5–6 per group). (C) Proportion of CD86 high CD206 low macrophages in TAMs ( n = 6 per group). (D) Proportion of CD86 low CD206 high macrophages among TAMs ( n = 6 per group). (E) Ratio of M1-like to M2-like macrophages ( n = 4 per group). (F) Proportion of CD3 lymphocytes in CD45 cells ( n = 5 per group). (G) Ratio of CD8 + /CD3 + T lymphocytes ( n = 5–7 per group). (H) Ratio of CD8 + to Treg ( n = 5–6 per group). (I) Flow cytometry analysis of PD-1 + CD8 + T, (J) PD-1 + CD4 + T cells, and (K) TIM3 + CD4 + T in the tumor ( n = 4–7 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: Astragalus-derived nano-agonist potentiates chemotherapy by reducing tumor-suppressive macrophages

    doi: 10.1016/j.apsb.2026.03.051

    Figure Lengend Snippet: Analysis of T lymphocyte phenotypes and macrophage subtypes in TME. (A) Proportion of CD45 cells in live cells ( n = 6 per group). (B) Ratio of macrophages to CD45 + T cells ( n = 5–6 per group). (C) Proportion of CD86 high CD206 low macrophages in TAMs ( n = 6 per group). (D) Proportion of CD86 low CD206 high macrophages among TAMs ( n = 6 per group). (E) Ratio of M1-like to M2-like macrophages ( n = 4 per group). (F) Proportion of CD3 lymphocytes in CD45 cells ( n = 5 per group). (G) Ratio of CD8 + /CD3 + T lymphocytes ( n = 5–7 per group). (H) Ratio of CD8 + to Treg ( n = 5–6 per group). (I) Flow cytometry analysis of PD-1 + CD8 + T, (J) PD-1 + CD4 + T cells, and (K) TIM3 + CD4 + T in the tumor ( n = 4–7 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA.

    Article Snippet: CD8 + T lymphocytes were extracted from the spleen of BALB/c mice with the CD8a microbeads kit (Miltenyi, 130-117-044, Bergisch Gladbach, Germany).

    Techniques: Flow Cytometry

    Analysis of T lymphocyte-related cytotoxic functions in the TME of 4T1 triple-negative breast cancer (TNBC) bearing mice treated with Vehicle, ADNPs, Cisplatin, or ADNPs-Cis. Proportion of (A) IFN- γ + CD8 + , (B) Granzyme B + CD8 + , (C) TNF- α + CD8 + ( n = 4–5 for each group). (D) Ki67 + CD8 + ( n = 5–6 for each group) in CD8 + T Cells. (E) HE staining analysis of CD8 expression in the tumor and IOD analysis ( n = 3, Scale bar = 50 μm). (F) IFN- γ + CD4 + ( n = 4–5 for each group). (G) Granzyme B + CD4 + ( n = 5 for each group). (H) TNF- α + CD4 + ( n = 5–6 for each group) among CD4 + T cells. (I) Schematic representation of the co-culture system for macrophages and CD8 + T lymphocytes. (J) ADNPs treated M2-like BMDMs reversed the proliferation of CD8 + T cells and (K) the statistics. ( n = 4 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: Astragalus-derived nano-agonist potentiates chemotherapy by reducing tumor-suppressive macrophages

    doi: 10.1016/j.apsb.2026.03.051

    Figure Lengend Snippet: Analysis of T lymphocyte-related cytotoxic functions in the TME of 4T1 triple-negative breast cancer (TNBC) bearing mice treated with Vehicle, ADNPs, Cisplatin, or ADNPs-Cis. Proportion of (A) IFN- γ + CD8 + , (B) Granzyme B + CD8 + , (C) TNF- α + CD8 + ( n = 4–5 for each group). (D) Ki67 + CD8 + ( n = 5–6 for each group) in CD8 + T Cells. (E) HE staining analysis of CD8 expression in the tumor and IOD analysis ( n = 3, Scale bar = 50 μm). (F) IFN- γ + CD4 + ( n = 4–5 for each group). (G) Granzyme B + CD4 + ( n = 5 for each group). (H) TNF- α + CD4 + ( n = 5–6 for each group) among CD4 + T cells. (I) Schematic representation of the co-culture system for macrophages and CD8 + T lymphocytes. (J) ADNPs treated M2-like BMDMs reversed the proliferation of CD8 + T cells and (K) the statistics. ( n = 4 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA.

    Article Snippet: CD8 + T lymphocytes were extracted from the spleen of BALB/c mice with the CD8a microbeads kit (Miltenyi, 130-117-044, Bergisch Gladbach, Germany).

    Techniques: Staining, Expressing, Co-Culture Assay

    ADNPs reprogram M2-like macrophages into an M1-like phenotype. (A) RT-PCR analysis of IL-6 , tnfα , cxcl9 , inos , tlr2 , and cd8 6 mRNA expression in M2-like macrophages treated with ADNPs for 6 and 12 h in vitro ( n = 3 per group). (B) ROS level in supernatants of BMDMs was detected by FACS in vitro . (C) Mean fluorescence intensity analysis of CD206, CD80, and CD86 of BMDMs in vitro ( n = 3 per group). (D) Concentration of TNF- α , IL-6, IL-10, and IL-8 in the supernatant of BMDMs treated with ADNPs in vitro ( n = 3–6 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA or t test.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: Astragalus-derived nano-agonist potentiates chemotherapy by reducing tumor-suppressive macrophages

    doi: 10.1016/j.apsb.2026.03.051

    Figure Lengend Snippet: ADNPs reprogram M2-like macrophages into an M1-like phenotype. (A) RT-PCR analysis of IL-6 , tnfα , cxcl9 , inos , tlr2 , and cd8 6 mRNA expression in M2-like macrophages treated with ADNPs for 6 and 12 h in vitro ( n = 3 per group). (B) ROS level in supernatants of BMDMs was detected by FACS in vitro . (C) Mean fluorescence intensity analysis of CD206, CD80, and CD86 of BMDMs in vitro ( n = 3 per group). (D) Concentration of TNF- α , IL-6, IL-10, and IL-8 in the supernatant of BMDMs treated with ADNPs in vitro ( n = 3–6 per group). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Data are presented as mean ± SEM and analyzed by one-way ANOVA or t test.

    Article Snippet: CD8 + T lymphocytes were extracted from the spleen of BALB/c mice with the CD8a microbeads kit (Miltenyi, 130-117-044, Bergisch Gladbach, Germany).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, In Vitro, Fluorescence, Concentration Assay