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Cell Signaling Technology Inc rabbit monoclonal anti cd4
Rabbit Monoclonal Anti Cd4, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+cd4+monoclonal+antibody/CD4+Rabbit+mAb/pm41619263-250-5-16
Average 95 stars, based on 46 article reviews

rabbit monoclonal anti cd4 - by Bioz Stars, 2026-10

95/100 stars

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Article Title: Tertiary lymphoid structure and decreased CD8 + T cell infiltration in minimally invasive adenocarcinoma
Article Snippet: Rabbit anti-CD4 monoclonal antibody (EP204) , CST , Cat#: 48274S.

Blocking Assay:

Article Title: Secreted phospholipase PLA2G12A-driven lysophospholipid signaling via lipolytic modification of extracellular vesicles facilitates pathogenic Th17 differentiation
Article Snippet: Paraffin-embedded sections were deparaffinized and antigen was activated with 20 μg/ml proteinase K (Merk Sigma-Aldrich). .. Endogenous peroxidase was inactivated by treatment with 3% ( w / v ) hydrogen peroxide solution (Fujifilm-Wako) for 5 min, and then blocked with Block Ace (DS Pharma Biomedical) for 1 h. The sections were incubated with anti-PLA2G12A antibody (clone #44) biotinylated with a Biotin-Labeling Kit (Dojindo) and rabbit anti-CD4 monoclonal antibody (D7D2Z; Cell Signaling) diluted at 1:1000 at room temperature for 1 h. After washing, the sections were incubated with APC streptavidin (BioLegend) and Alexa Flour 488-conjugated F(ab’) 2 -goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody (A-11070, Thermo Fisher Scientific) diluted at 1:1000 at room temperature for 1 h. After washing, the sections were sealed with VECTASHIELD mounting medium with 4’,6-diamidino-2-phenylindole (DAPI) (Vector Laboratories). .. RIPA Buffer (Nacalai Tesque) mixed with protease inhibitors (cOmplete Protease Inhibitor Cocktail tablets) and phosphatase inhibitors (PhosSTOP Phosphatase Inhibitor Cocktail tablets) (both from Roche) was added to cell lysates.

Incubation:

Image Search Results

MiR-138 directly targets STAT1 and indirectly regulates antitumor immunosurveillance (A) Tumor growth of syngeneic mice injected with TuD-138 or TuD-NC Neuro2a cells ( n = 10–13 per group). (B) Tumor growth in nude mice ( n = 10 per group). (C) Flow cytometric quantification of tumor-infiltrating CD4 + and CD8 + T cells ( n = 3). (D) Tumor growth in mice depleted of lymphocytes before implantation ( n = 5 per group). (E) STAT1 mRNA expression in tumors measured by RT-qPCR ( n = 3). (F) STAT1 protein expression in tumor lysates by western blotting. (G) Immunohistochemistry of STAT1 in tumor sections (scale bar, 50 μm). (H) (i) Predicted miR-138 binding sites within STAT1 3′UTR and corresponding mutants. (ii) Dual-luciferase assay in Neuro2a-TuD138 or Neuro2a-TuD NC cells transfected with wild-type or mutant STAT1 3′UTR vectors. (I) Flow cytometric analysis of MHC class I expression in cells and tumors. (i) Representative histograms. (ii) Quantified mean fluorescence intensity (MFI). Data are shown as mean. Bars, SEM. Statistical analysis: Student’s t test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Journal: Molecular Therapy Oncology

Article Title: MicroRNA-138 regulation promotes the efficacy of oncolytic herpes virus G47Δ for malignant brain tumor therapy

doi: 10.1016/j.omton.2025.201108

Figure Lengend Snippet: MiR-138 directly targets STAT1 and indirectly regulates antitumor immunosurveillance (A) Tumor growth of syngeneic mice injected with TuD-138 or TuD-NC Neuro2a cells ( n = 10–13 per group). (B) Tumor growth in nude mice ( n = 10 per group). (C) Flow cytometric quantification of tumor-infiltrating CD4 + and CD8 + T cells ( n = 3). (D) Tumor growth in mice depleted of lymphocytes before implantation ( n = 5 per group). (E) STAT1 mRNA expression in tumors measured by RT-qPCR ( n = 3). (F) STAT1 protein expression in tumor lysates by western blotting. (G) Immunohistochemistry of STAT1 in tumor sections (scale bar, 50 μm). (H) (i) Predicted miR-138 binding sites within STAT1 3′UTR and corresponding mutants. (ii) Dual-luciferase assay in Neuro2a-TuD138 or Neuro2a-TuD NC cells transfected with wild-type or mutant STAT1 3′UTR vectors. (I) Flow cytometric analysis of MHC class I expression in cells and tumors. (i) Representative histograms. (ii) Quantified mean fluorescence intensity (MFI). Data are shown as mean. Bars, SEM. Statistical analysis: Student’s t test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Anti-CD8 (clone 53–6.72) and anti-CD4 (clone GK1.5) monoclonal antibodies were obtained from BioX Cell (Lebanon, NH, USA).

Techniques: Injection, Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Binding Assay, Luciferase, Transfection, Mutagenesis, Fluorescence

Donor-type CD4 + T cells from livers of No GVHD or cGVHD mice on day 60 after HCT were sorted and subjected to scRNA-Seq ( N = 3). a Uniform manifold approximation and projection (UMAP) plots. Workflow: Created in BioRender. (Kong, W. (2026) https://BioRender.com/6hbegtm ) . b Feature plots show expression of genes related to quiescence, activation, migration, stemness, tissue residency and proinflammation. c Feature plots display T cell progenitor signature. d KEGG pathway of DEGs from each cluster. e Proportions of CD4 + T cell subsets defined by clustering. f Representative panel showed four Tm subsets with TCF1-EGFP ( Tcf7 -EGFP) versus CD69 in the liver of No GVHD and cGVHD mice at day 60 after HCT. g Comparison of protein expression among four CD4 + Tm subsets identified with Ly108 vs CD69, N = 4–6. h %IFN-γ + GM-CSF + cells in each Tm subset after in vitro stimulation with PMA and ionomycin. N = 6. i Percentages and yields of four Tm subsets, N = 5.Data presented as mean ± SEM from two repeated experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, as determined by one-way ANOVA ( g , h ) and two-way ANOVA ( i ).

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: Donor-type CD4 + T cells from livers of No GVHD or cGVHD mice on day 60 after HCT were sorted and subjected to scRNA-Seq ( N = 3). a Uniform manifold approximation and projection (UMAP) plots. Workflow: Created in BioRender. (Kong, W. (2026) https://BioRender.com/6hbegtm ) . b Feature plots show expression of genes related to quiescence, activation, migration, stemness, tissue residency and proinflammation. c Feature plots display T cell progenitor signature. d KEGG pathway of DEGs from each cluster. e Proportions of CD4 + T cell subsets defined by clustering. f Representative panel showed four Tm subsets with TCF1-EGFP ( Tcf7 -EGFP) versus CD69 in the liver of No GVHD and cGVHD mice at day 60 after HCT. g Comparison of protein expression among four CD4 + Tm subsets identified with Ly108 vs CD69, N = 4–6. h %IFN-γ + GM-CSF + cells in each Tm subset after in vitro stimulation with PMA and ionomycin. N = 6. i Percentages and yields of four Tm subsets, N = 5.Data presented as mean ± SEM from two repeated experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, as determined by one-way ANOVA ( g , h ) and two-way ANOVA ( i ).

Article Snippet: cGVHD mice were treated with intraperitoneal (i.p.) anti-mouse CD4 mAbs (Bio X Cell, Clone GK1.5) weekly at 500 μg/mouse from day 29 to 50 after HCT or from day 29 to 99 after HCT.

Techniques: Expressing, Activation Assay, Migration, Comparison, In Vitro

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: Pooled donor-type CD4 + T cells from the lung at day 60 after HCT were sorted and processed for bulk combined RNA-Seq and ATAC-Seq. N = 3. a PCA plot of the four Tm subsets. b Heatmap of sample distances showing the gene profile similarity between the four Tm subsets. c Heatmap shows all DEGs in the four Tm subsets with clusters of Pearson correlation. d Bubble graph displays the top five enriched KEGG pathways for each cluster from ( c ). e Volcano plot shows top twenty upregulated or downregulated DEGs annotated in the comparison between Tsm and Trmp and between Tint and Trm cells. f Duplicate tSNE maps show the chromatin accessibility profiles of the four Tm subsets. g Volcano plots comparing specific peaks of transcription factor motif enrichment of Tm subsets: Tsm vs Trmp, Trmp vs Trm, Tsm vs Tint and Trm vs Tint. h ATAC-seq tracks at the loci of Slamf6 (Ly108), Tcf7, Cxcr5, Bhlhe40, Tbx21 (T-bet) and Klrk1 (NKG2D).

Techniques: RNA Sequencing, Comparison

a Single-cell trajectory of the liver CD4 + T cell subsets with pseudotime analysis. b UMAP plot displaying clonal TRB counts of donor-type CD4 + T cells from the liver. c Scatter plot comparing TRB clonotype overlap and expansion. d Clonality index values were analyzed and compared among CD4 + T cell subsets from the liver of cGVHD. e UMAP plot shows cluster distribution for six representatives TCR clonotypes. f Bar plots show numbers of twelve expanded TCR clonotypes in different Tm clusters (Black) and total numbers of cells with each clonotype (Blue). g Heatmap shows differentially expressed genes among the top 20 expanded T clonotypes.

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: a Single-cell trajectory of the liver CD4 + T cell subsets with pseudotime analysis. b UMAP plot displaying clonal TRB counts of donor-type CD4 + T cells from the liver. c Scatter plot comparing TRB clonotype overlap and expansion. d Clonality index values were analyzed and compared among CD4 + T cell subsets from the liver of cGVHD. e UMAP plot shows cluster distribution for six representatives TCR clonotypes. f Bar plots show numbers of twelve expanded TCR clonotypes in different Tm clusters (Black) and total numbers of cells with each clonotype (Blue). g Heatmap shows differentially expressed genes among the top 20 expanded T clonotypes.

Techniques: Single Cell

a Tsm, Trmp and Trm subsets were sorted from the liver and lung of cGVHD mice at day 30 after HCT and adoptively transferred into cGVHD recipients at day 14 after HCT. Adoptively transferred CD45.1 + CD4 + Tm subsets were analyzed 14 days later, as shown in the diagram. b Recovery of each injected Tm subset in the liver and lung 14 days after adoptive transfer, N = 4–8. c Percentages of Tsm, Trmp and Trm among cells derived from each injected Tm subsets are shown and calculated, N = 4–5. d %Tox positive T cells among injected CD45.1 + Tm subsets, N = 4 per group. e CD4 + Tm subsets were sorted from the liver and lung of primary cGVHD mice at day 30 after HCT and adoptively transferred into secondary cGVHD recipients established with Rag1 −/− -BM plus 0.1 M CD8 + T cells at day 15 after HCT, %baseline body weight and clinical GVHD score are shown, N = 8. f Representative Masion’ Trichrome panel (magnification, x100, N = 4) and pathology score are displayed. g Yields of Tsm, Trmp and Trm cells in the liver and lung derived from injected CD45.1 + Tm subsets, N = 5 or 6 per group. h %baseline bodyweight and %survival among cGVHD mice treated with anti-CD4 mAbs or control IgG weekly from day 29 to 50 after HCT. N = 7 or 10 per group. i Representative Masion’ Trichrome panel (magnification, x200) and pathology score on day 60 of IgG control and aCD4 mAbs group, ~day75 in the cGVHD recurrence group after HCT, N = 9, 4 or 5 per group. j Yields of Tm subsets in the liver and lungs of cGVHD recipients treated in ( h ). k %baseline bodyweight and %survival among cGVHD mice treated with aCD4 mAbs or control IgG weekly from day 29 to 99 after HCT. N = 13 or 14. Data presented as mean ± SEM from at least 2 replicates. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by one-way ANOVA ( b , d , f ), unpaired two-tailed t-test ( e , h , k ), log-rank test comparison of survival ( h , k ) and two-way ANOVA ( g , i , j ).

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: a Tsm, Trmp and Trm subsets were sorted from the liver and lung of cGVHD mice at day 30 after HCT and adoptively transferred into cGVHD recipients at day 14 after HCT. Adoptively transferred CD45.1 + CD4 + Tm subsets were analyzed 14 days later, as shown in the diagram. b Recovery of each injected Tm subset in the liver and lung 14 days after adoptive transfer, N = 4–8. c Percentages of Tsm, Trmp and Trm among cells derived from each injected Tm subsets are shown and calculated, N = 4–5. d %Tox positive T cells among injected CD45.1 + Tm subsets, N = 4 per group. e CD4 + Tm subsets were sorted from the liver and lung of primary cGVHD mice at day 30 after HCT and adoptively transferred into secondary cGVHD recipients established with Rag1 −/− -BM plus 0.1 M CD8 + T cells at day 15 after HCT, %baseline body weight and clinical GVHD score are shown, N = 8. f Representative Masion’ Trichrome panel (magnification, x100, N = 4) and pathology score are displayed. g Yields of Tsm, Trmp and Trm cells in the liver and lung derived from injected CD45.1 + Tm subsets, N = 5 or 6 per group. h %baseline bodyweight and %survival among cGVHD mice treated with anti-CD4 mAbs or control IgG weekly from day 29 to 50 after HCT. N = 7 or 10 per group. i Representative Masion’ Trichrome panel (magnification, x200) and pathology score on day 60 of IgG control and aCD4 mAbs group, ~day75 in the cGVHD recurrence group after HCT, N = 9, 4 or 5 per group. j Yields of Tm subsets in the liver and lungs of cGVHD recipients treated in ( h ). k %baseline bodyweight and %survival among cGVHD mice treated with aCD4 mAbs or control IgG weekly from day 29 to 99 after HCT. N = 13 or 14. Data presented as mean ± SEM from at least 2 replicates. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by one-way ANOVA ( b , d , f ), unpaired two-tailed t-test ( e , h , k ), log-rank test comparison of survival ( h , k ) and two-way ANOVA ( g , i , j ).

Techniques: Injection, Adoptive Transfer Assay, Derivative Assay, Control, Two Tailed Test, Comparison

a Donor derived CD11b + CD11c + APCs gated from H2Kb + TCRβ − cells from the lung of No GVHD and cGVHD mice were examined at day 60 after HCT, expression of MHCII from APCs was also measured, N = 4. b , c cGVHD mice were established with CD45.2 + WT or MHCII −/− CD45.2 + BALB/c recipients given C57BL/6 CD45.2 + WT-BM or MHCII −/− - BM plus CD45.1 + WT-T cells. Four Tm subsets among injected donor-type (CD45.1 + ) CD4 + T cells from the lung ( b ) and liver ( c ) were examined on days 7 ( N = 3 or 4 per group) and 60 after HCT ( N = 6, 4 or 6 per group). d Proinflammatory IFN-γ and GM-CSF production among Trm cells was measured on day 60 after HCT ( N = 6 per group). e BALB/c recipients were established with WT-T plus WT or IFNγR −/− TCD-BM. %survival and %baseline bodyweight. N = 10–15. f Representative Masion’ trichrome staining panel and pathology scores are shown (magnification, x200, N = 4). g Representative contour plots, percentages and yields of the four Tm subsets were analyzed on day 60 after HCT, N = 4 ~ 6. h Proinflammatory IFN-γ and GM-CSF production among CD4 + Tm cells, N = 4. i Percentages of CD11b + CD11c + APCs were also compared, and MHCII expression was analyzed, N = 4 ~ 6. Data presented as mean ± SEM from at least 2 replicates. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by two-way ANOVA ( a , b , f , g ), one-way ANOVA ( d ), log-rank test comparison of survival ( e ) and unpaired two-tailed t-test ( e , h , i ).

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: a Donor derived CD11b + CD11c + APCs gated from H2Kb + TCRβ − cells from the lung of No GVHD and cGVHD mice were examined at day 60 after HCT, expression of MHCII from APCs was also measured, N = 4. b , c cGVHD mice were established with CD45.2 + WT or MHCII −/− CD45.2 + BALB/c recipients given C57BL/6 CD45.2 + WT-BM or MHCII −/− - BM plus CD45.1 + WT-T cells. Four Tm subsets among injected donor-type (CD45.1 + ) CD4 + T cells from the lung ( b ) and liver ( c ) were examined on days 7 ( N = 3 or 4 per group) and 60 after HCT ( N = 6, 4 or 6 per group). d Proinflammatory IFN-γ and GM-CSF production among Trm cells was measured on day 60 after HCT ( N = 6 per group). e BALB/c recipients were established with WT-T plus WT or IFNγR −/− TCD-BM. %survival and %baseline bodyweight. N = 10–15. f Representative Masion’ trichrome staining panel and pathology scores are shown (magnification, x200, N = 4). g Representative contour plots, percentages and yields of the four Tm subsets were analyzed on day 60 after HCT, N = 4 ~ 6. h Proinflammatory IFN-γ and GM-CSF production among CD4 + Tm cells, N = 4. i Percentages of CD11b + CD11c + APCs were also compared, and MHCII expression was analyzed, N = 4 ~ 6. Data presented as mean ± SEM from at least 2 replicates. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by two-way ANOVA ( a , b , f , g ), one-way ANOVA ( d ), log-rank test comparison of survival ( e ) and unpaired two-tailed t-test ( e , h , i ).

Techniques: Derivative Assay, Expressing, Injection, Staining, Comparison, Two Tailed Test

a Representative patterns, percentages and numbers of TCF1 + T-bet lo Tsm and TCF1 lo T-bet hi Teff cells from the PBL of allo-HCT patients with No-GVHD and active cGVHD are shown and compared. N = 7. b Representative histograms show protein expression with ( c ) heatmap displaying relative expression among Tm subsets. d Tertiary lymphoid-like structures are shown with CD3, CD45RA, CD68, CD20, Collagen I and CD11c. e Merged images show staining of CD4, CD3, CD45RA, Foxp3 and DNA and f staining of IFN-γ, GM-CSF and CD4, and percentages of IFN-γ and GM-CSF producing CD4 + Tm cells and Treg cells were measured. g Merged image shows staining of CD69, Ly108 and CD4 in the liver of patients with active cGVHD. h Representative images of CCR7 − CD69 + CD4 + CD3 + T cells of liver sections from cGVHD patients. i Representative images of TCF1 and CD69 expression among CD4 + CD45RA − cells of liver sections from cGVHD patients, N = 4. Data represented as means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by two-way ANOVA ( a ), one-way ANOVA ( g,i ) and unpaired two-tailed t-test (h) .

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: a Representative patterns, percentages and numbers of TCF1 + T-bet lo Tsm and TCF1 lo T-bet hi Teff cells from the PBL of allo-HCT patients with No-GVHD and active cGVHD are shown and compared. N = 7. b Representative histograms show protein expression with ( c ) heatmap displaying relative expression among Tm subsets. d Tertiary lymphoid-like structures are shown with CD3, CD45RA, CD68, CD20, Collagen I and CD11c. e Merged images show staining of CD4, CD3, CD45RA, Foxp3 and DNA and f staining of IFN-γ, GM-CSF and CD4, and percentages of IFN-γ and GM-CSF producing CD4 + Tm cells and Treg cells were measured. g Merged image shows staining of CD69, Ly108 and CD4 in the liver of patients with active cGVHD. h Representative images of CCR7 − CD69 + CD4 + CD3 + T cells of liver sections from cGVHD patients. i Representative images of TCF1 and CD69 expression among CD4 + CD45RA − cells of liver sections from cGVHD patients, N = 4. Data represented as means ± SEMs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, as determined by two-way ANOVA ( a ), one-way ANOVA ( g,i ) and unpaired two-tailed t-test (h) .

Techniques: Expressing, Staining, Two Tailed Test

Based on distinct transcriptional and epigenomic features and flow cytometry of Ly108 versus CD69, donor graft CD4 + T-derived memory T cells (Tm) in the chronic GVHD target tissues can be divided into four subsets: Ly108 + CD69 − Tsm, Ly108 + CD69 + Trmp, Ly108 − CD69 + Trm, and Ly108 − CD69 − Tint. Trm are the critical effectors that mediate cGVHD, and Tsm are required for maintaining the Tm pool. Tsm differentiation into Trmp and then to Trm in a MHCII/TCR-dependent manner and produces biased clonal expansion of Trm cells with strong proinflammatory characteristics. TCF1/BCL6 and BHLHE40 differentially regulate the stemness and differentiation of Tsm cells. Solid lines with arrow mean strong indication and dashed lines with arrow means potential indication. Created in BioRender. Kong, W. (2026) https://BioRender.com/2extxe5 .

Journal: Nature Communications

Article Title: Stem-like memory-T maintenance and differentiation into tissue-resident T cells sustain chronic graft-versus-host disease in mice

doi: 10.1038/s41467-026-69975-z

Figure Lengend Snippet: Based on distinct transcriptional and epigenomic features and flow cytometry of Ly108 versus CD69, donor graft CD4 + T-derived memory T cells (Tm) in the chronic GVHD target tissues can be divided into four subsets: Ly108 + CD69 − Tsm, Ly108 + CD69 + Trmp, Ly108 − CD69 + Trm, and Ly108 − CD69 − Tint. Trm are the critical effectors that mediate cGVHD, and Tsm are required for maintaining the Tm pool. Tsm differentiation into Trmp and then to Trm in a MHCII/TCR-dependent manner and produces biased clonal expansion of Trm cells with strong proinflammatory characteristics. TCF1/BCL6 and BHLHE40 differentially regulate the stemness and differentiation of Tsm cells. Solid lines with arrow mean strong indication and dashed lines with arrow means potential indication. Created in BioRender. Kong, W. (2026) https://BioRender.com/2extxe5 .

Techniques: Flow Cytometry, Derivative Assay

Representative immunofluorescence micrographs and quantitative analysis of CD4 + T−cell infiltration in tumor tissue.** Images captured at 200× magnification; scale bar = 50 µm. Data represent **mean ± SEM of % positive area per animal** (n = 6 mice/group). Fold change relative to PCG displayed on bars. *P < 0.05 vs PCG; one−way ANOVA.*.

Journal: Frontiers in Oncology

Article Title: Immuno−oncological effects of aerobic exercise combined with anti−PD−L1 antibody blockade in a murine breast cancer model

doi: 10.3389/fonc.2026.1671244

Figure Lengend Snippet: Representative immunofluorescence micrographs and quantitative analysis of CD4 + T−cell infiltration in tumor tissue.** Images captured at 200× magnification; scale bar = 50 µm. Data represent **mean ± SEM of % positive area per animal** (n = 6 mice/group). Fold change relative to PCG displayed on bars. *P < 0.05 vs PCG; one−way ANOVA.*.

Article Snippet: Antibodies used in this study were as follows: InVivoMAb anti−mouse PD−L1 (B7−H1; clone 10F.9G2TM, Bio X Cell, USA; catalog no. BE0101), rabbit anti−mouse CD4 monoclonal antibody (clone D7D2Z, Cell Signaling Technology, USA; catalog no. 25229; UniProt ID: P06332 ), rabbit anti−mouse CD8α monoclonal antibody (clone D4W2Z, Cell Signaling Technology, USA; catalog no. 98941; UniProt ID: P01731 ), and FITC−conjugated goat anti−rabbit IgG (H+L) secondary antibody (Elabscience, USA; catalog no. E−AB−1014), RPMI−1640 medium (Gibco, Thermo Fisher Scientific, USA; catalog no. 11875−093), fetal bovine serum (FBS; Gibco, USA; catalog no. 10082−147), penicillin–streptomycin (Gibco, USA; catalog no. 15140−122), trypsin−EDTA (Gibco, USA; catalog no. 25200−056) and phosphate−buffered saline (PBS; Gibco, USA; catalog no. 10010−023).

Techniques: Immunofluorescence

Quantitative analysis of CD4 + immune cell infiltration within tumor tissue. Data represent mean ± SEM of % CD4 + −positive area (n = 6 mice/group), normalized to PCG (fold change shown on bars). One−way ANOVA; P < 0.05.*. * The significance of the EIA with the PCG ** The significance of the EIE+AE group with the PCG.

Journal: Frontiers in Oncology

Article Title: Immuno−oncological effects of aerobic exercise combined with anti−PD−L1 antibody blockade in a murine breast cancer model

doi: 10.3389/fonc.2026.1671244

Figure Lengend Snippet: Quantitative analysis of CD4 + immune cell infiltration within tumor tissue. Data represent mean ± SEM of % CD4 + −positive area (n = 6 mice/group), normalized to PCG (fold change shown on bars). One−way ANOVA; P < 0.05.*. * The significance of the EIA with the PCG ** The significance of the EIE+AE group with the PCG.

Techniques:

Ratio of CD8 + /CD4 + T−cell infiltration within tumor tissue. Values represent mean ± SEM (n = 6 mice/group) based on percentage of positive area quantified by ImageJ 1.53a. Statistical analysis was performed using one−way ANOVA (P < 0.05). Higher ratio values indicate predominance of cytotoxic CD8 + T−cells relative to helper CD4 + T−cells.

Journal: Frontiers in Oncology

Article Title: Immuno−oncological effects of aerobic exercise combined with anti−PD−L1 antibody blockade in a murine breast cancer model

doi: 10.3389/fonc.2026.1671244

Figure Lengend Snippet: Ratio of CD8 + /CD4 + T−cell infiltration within tumor tissue. Values represent mean ± SEM (n = 6 mice/group) based on percentage of positive area quantified by ImageJ 1.53a. Statistical analysis was performed using one−way ANOVA (P < 0.05). Higher ratio values indicate predominance of cytotoxic CD8 + T−cells relative to helper CD4 + T−cells.

Techniques:

CD4+ and CD8+ in tumor tissue changes in between the EIE, EIA and EIE+A groups compared to the EIC.

Journal: Frontiers in Oncology

Article Title: Immuno−oncological effects of aerobic exercise combined with anti−PD−L1 antibody blockade in a murine breast cancer model

doi: 10.3389/fonc.2026.1671244

Figure Lengend Snippet: CD4+ and CD8+ in tumor tissue changes in between the EIE, EIA and EIE+A groups compared to the EIC.

Techniques:

Quantitative analysis of CD4 + cell infiltration in tumor tissue. Data expressed as mean ± SEM % positive area (n = 6 mice/group) normalized to PCG. Fold change vs PCG shown on bars. P < 0.05 vs PCG by one−way ANOVA.

Journal: Frontiers in Oncology

Article Title: Immuno−oncological effects of aerobic exercise combined with anti−PD−L1 antibody blockade in a murine breast cancer model

doi: 10.3389/fonc.2026.1671244

Figure Lengend Snippet: Quantitative analysis of CD4 + cell infiltration in tumor tissue. Data expressed as mean ± SEM % positive area (n = 6 mice/group) normalized to PCG. Fold change vs PCG shown on bars. P < 0.05 vs PCG by one−way ANOVA.

Techniques:

a , Representative flow cytometry plots of OVA-tetramer+ CD8+ T cells from Day 12 tdLN of mice pre-treated with or without anti-CD4 depleting antibodies, and with control IgG or anti-PD-L1/PD-L2 according to Fig. experimental scheme. b, c , Mean fluorescence intensity of TCF-1 ( b ) and SLAMF6 ( c ) of OVA-tetramer+ TCF-1+ T SL normalized to naïve (CD44- TCF-1+ PD-1-) CD8+ T cells. d, e , Quantification of OVA-tetramer+ SLAMF6+ T SL from Day 12 tdLN ( d ) and re-scaled for each group (non-depleted and CD4-depleted) to show the relative extent of SLAMF6+ T SL reduction after checkpoint treatment ( e ). f, g , Standardized log transformed imputed TCR affinity index ( f ) and mean imputed affinity index ( g ) of each treatment group, normalized to non-CD4-depleted, IgG-treated SLAMF6+ T SL from the same experimental cohort. Each data point represents the mean subset value from each animal. h , Tumour volume measured at the experimental end point at Day 12. Data from 3 independent experiments (n = 9 for non-depleted IgG and aPD-L1/2; n = 6 for CD4-depleted IgG; n = 10 for CD4-depleted aPD-L1/2). Error bars: means ± s.d. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; unpaired two-tailed t -test in ( e ), one-way ANOVA with Tukey’s multiple comparisons in all others.

Journal: Nature

Article Title: Inhibitory PD-1 axis maintains high-avidity stem-like CD8 + T cells

doi: 10.1038/s41586-025-09440-x

Figure Lengend Snippet: a , Representative flow cytometry plots of OVA-tetramer+ CD8+ T cells from Day 12 tdLN of mice pre-treated with or without anti-CD4 depleting antibodies, and with control IgG or anti-PD-L1/PD-L2 according to Fig. experimental scheme. b, c , Mean fluorescence intensity of TCF-1 ( b ) and SLAMF6 ( c ) of OVA-tetramer+ TCF-1+ T SL normalized to naïve (CD44- TCF-1+ PD-1-) CD8+ T cells. d, e , Quantification of OVA-tetramer+ SLAMF6+ T SL from Day 12 tdLN ( d ) and re-scaled for each group (non-depleted and CD4-depleted) to show the relative extent of SLAMF6+ T SL reduction after checkpoint treatment ( e ). f, g , Standardized log transformed imputed TCR affinity index ( f ) and mean imputed affinity index ( g ) of each treatment group, normalized to non-CD4-depleted, IgG-treated SLAMF6+ T SL from the same experimental cohort. Each data point represents the mean subset value from each animal. h , Tumour volume measured at the experimental end point at Day 12. Data from 3 independent experiments (n = 9 for non-depleted IgG and aPD-L1/2; n = 6 for CD4-depleted IgG; n = 10 for CD4-depleted aPD-L1/2). Error bars: means ± s.d. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; unpaired two-tailed t -test in ( e ), one-way ANOVA with Tukey’s multiple comparisons in all others.

Article Snippet: For depletion of CD4 + T cells, mice were intraperitoneally injected with 100 μg anti-mouse CD4 monoclonal antibody (GK1.5, BioXCell) 3 days and 1 day before tumour induction.

Techniques: Flow Cytometry, Control, Fluorescence, Transformation Assay, Two Tailed Test

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