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).

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: RNA Sequencing, Comparison

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.

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: Single Cell

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 ).

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

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 ).

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

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) .

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, Staining, Two Tailed Test

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 .

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: Flow Cytometry, Derivative Assay

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Expansion of T cells induced by rDEV-dH5/H7 and vDEV. Four-week-old ducks were inoculated i.m. with two doses of 10 5 TCID 50 of rDEV-dH5/H7 or vDEV at a 3-week interval. The percentages of ( A ) CD3 + T cells, ( B ) CD3 + CD8 + T cells, and ( C ) CD3 + CD4 + T cells among the PBMCs were analyzed by flow cytometry. Data are presented as means and plotted with connecting lines. Each data point represents one sample value. The red triangles represent the time points of inoculation.

Article Snippet: The antibody cocktail included an FITC-conjugated anti-CD3 mAb (CD3-12; Abcam, ab11089) together with a PE-conjugated anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or a PE-conjugated anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479), which were labeled with a PE/R-phycoerythrin Conjugation Kit (Abcam, ab102918), respectively.

Techniques: Flow Cytometry

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Specific CD4 + T-cell responses induced by rDEV-dH5/H7 and vDEV. ( A ) Timeline of inoculation and sample testing. Arrows indicate inoculation, and red circles indicate bleeds, tests, and post-vaccination time points. ( B ) Frequency of specific CD3 + CD4 + T cells expressing IFN-γ + induced by DEV virion. ( C–F ) Frequency of HA-specific CD3 + CD4 + T cells expressing IFN-γ + . PBMCs were stimulated with single inactivated virus antigens: GZ/S4184 (H5N6), LN/SD007 (H5N1), GX/SD098 (H7N9), or a mixture of antigens of these three influenza virus antigens; CD3 + CD4 + IFN-γ + T cells were then quantified. ( G and H ) Uncorrelated control. CD3 + CD4 + IFN-γ + T cells were quantified after PBMCs were stimulated with WSN (H1N1) or La Sota (NDV). Data are presented as means in histograms. Each data point represents one sample value. Statistical significance: * P < 0.05, ** P <0.01, and *** P < 0.001. P values were determined using a two-tailed unpaired Student’s t test. ( I ) Representative gating strategy for HA-specific CD3 + CD4 + T cells expressing IFN-γ + detected at 31 days post-prime vaccination, following stimulation with the influenza virus antigens mixture, RPMI 1640, and PMA plus ionomycin served as negative and positive controls, respectively.

Article Snippet: The antibody cocktail included an FITC-conjugated anti-CD3 mAb (CD3-12; Abcam, ab11089) together with a PE-conjugated anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or a PE-conjugated anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479), which were labeled with a PE/R-phycoerythrin Conjugation Kit (Abcam, ab102918), respectively.

Techniques: Expressing, Virus, Control, Two Tailed Test

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Expansion of T cells induced by rDEV-dH5/H7 and vDEV. Four-week-old ducks were inoculated i.m. with two doses of 10 5 TCID 50 of rDEV-dH5/H7 or vDEV at a 3-week interval. The percentages of ( A ) CD3 + T cells, ( B ) CD3 + CD8 + T cells, and ( C ) CD3 + CD4 + T cells among the PBMCs were analyzed by flow cytometry. Data are presented as means and plotted with connecting lines. Each data point represents one sample value. The red triangles represent the time points of inoculation.

Article Snippet: The PBMCs were plated in 1.5 mL centrifuge tubes (10 6 cells/tube) and fixed with 100 μL of 0.3% paraformaldehyde for 20 min at room temperature and subsequently stained for 30 min at room temperature with either mouse anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or mouse anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479) diluted in PBS containing 0.05% Tween-20.

Techniques: Flow Cytometry

Journal: Journal of Virology

Article Title: Recombinant duck enteritis virus harboring the hemagglutinin genes of influenza virus rapidly induces specific cellular immunity in ducks

doi: 10.1128/jvi.02014-25

Figure Lengend Snippet: Specific CD4 + T-cell responses induced by rDEV-dH5/H7 and vDEV. ( A ) Timeline of inoculation and sample testing. Arrows indicate inoculation, and red circles indicate bleeds, tests, and post-vaccination time points. ( B ) Frequency of specific CD3 + CD4 + T cells expressing IFN-γ + induced by DEV virion. ( C–F ) Frequency of HA-specific CD3 + CD4 + T cells expressing IFN-γ + . PBMCs were stimulated with single inactivated virus antigens: GZ/S4184 (H5N6), LN/SD007 (H5N1), GX/SD098 (H7N9), or a mixture of antigens of these three influenza virus antigens; CD3 + CD4 + IFN-γ + T cells were then quantified. ( G and H ) Uncorrelated control. CD3 + CD4 + IFN-γ + T cells were quantified after PBMCs were stimulated with WSN (H1N1) or La Sota (NDV). Data are presented as means in histograms. Each data point represents one sample value. Statistical significance: * P < 0.05, ** P <0.01, and *** P < 0.001. P values were determined using a two-tailed unpaired Student’s t test. ( I ) Representative gating strategy for HA-specific CD3 + CD4 + T cells expressing IFN-γ + detected at 31 days post-prime vaccination, following stimulation with the influenza virus antigens mixture, RPMI 1640, and PMA plus ionomycin served as negative and positive controls, respectively.

Article Snippet: The PBMCs were plated in 1.5 mL centrifuge tubes (10 6 cells/tube) and fixed with 100 μL of 0.3% paraformaldehyde for 20 min at room temperature and subsequently stained for 30 min at room temperature with either mouse anti-duck CD4 mAb (Du CD4-2; Bio-Rad, MCA2478) or mouse anti-duck CD8 mAb (Du CD8-1; Bio-Rad, MCA2479) diluted in PBS containing 0.05% Tween-20.

Techniques: Expressing, Virus, Control, Two Tailed Test

Journal: Nature Communications

Article Title: Circadian fluctuation of soluble CD26 dictates the impact of the timing of cord blood transplantation on acute graft-versus-host disease

doi: 10.1038/s41467-026-68958-4

Figure Lengend Snippet: a Experimental design. Three doses of Sitagliptin (30 mg/kg, DPPIV inhibitor) or an equal volume of PBS were intraperitoneally given to ZT5 mice before human cord blood-derived mononuclear cells (CBMCs) infusions. b Representative images of PBS-treated (top) and Sitagliptin-treated (bottom) ZT5 recipients showing phenotypic differences on day 21 after CBT. c – e aGVHD scores ( c , P = 0.003), survival rates ( d , P = 0.030) and body weights ( e , P = 0.624) of recipients posttransplantation ( n = 8 mice for the control group; n = 5 for the Sitagliptin group). f , g Histopathologic analysis of liver, lung, spleen and colon sections on day 21 after CBT. Representative H&E-stained images ( f ) and quantification results ( g ) ( n = 24-30 images from 4-6 mice per group) Two-sided unpaired t-tests were used for comparisons. The exact P values were: P < 0.001 for liver, lung, and spleen; P = 0.850 for colon. h – k The infiltration of CD4 + T cells and CD8 + T cells in liver, lung, spleen and colon sections on day 21 after transplantation. Representative immunohistochemistry images and quantification results of staining with anti-human CD4 ( h , i ) or CD8 ( j , k ) polyclonal antibodies ( n = 3 mice per group; 24 fields of view were analyzed per organ). For CD4⁺ T cell infiltration, statistical comparisons yielded P < 0.001 for liver and lung, P = 0.002 for spleen, and P = 0.285 for colon; For CD8⁺ T cell infiltration, statistical comparisons yielded P < 0.001 for liver, lung and spleen, and P > 0.999 for colon. AOD, average optical density. Scale bars, 100 μm. The data are presented as the means ± SEMs and were analyzed by unpaired t tests followed by Bonferroni-Dunn correction ( g , i and k ), two-way ANOVA followed by the Bonferroni post hoc correction ( c and e ), and the log-rank test ( d ). Source data are provided as a Source Data file.

Article Snippet: Immunostaining was carried out via the following antibodies: anti-human CD4 (ab133616, Abcam, dilution 1:500), anti-human CD8 (ZA-0508, ZSGB-BIO, dilution 1:100), anti-mouse CD4 (25229S, CST, dilution 1:100), and anti-mouse CD8 (A23305PM, ABclonal, dilution 1:300).

Techniques: Derivative Assay, Control, Staining, Transplantation Assay, Immunohistochemistry

Journal: Nature Communications

Article Title: Circadian fluctuation of soluble CD26 dictates the impact of the timing of cord blood transplantation on acute graft-versus-host disease

doi: 10.1038/s41467-026-68958-4

Figure Lengend Snippet: PBMCs from healthy donors were cultured with PHA-L (control group), PHA-L plus CD26/DPPIV (sCD26 group), or PHA-L plus Sitagliptin (DPPIV inhibitor, DPPIVi group) for 72 h. a Experimental design. PBMCs from healthy donors were cultured with PHA-L (control group), PHA-L plus CD26/DPPIV (sCD26 group), or PHA-L plus Sitagliptin (DPPIV inhibitor, DPPIVi group) for 72 h. b , c Representative plots (left) and quantified percentages (right) of CD86 + ( b ) and CD80 + ( c ) CD14 + cells after 72 h of PBMCs coculture in different groups (pooled data from 8 healthy donors across four independent experiments). d – i Representative plots (left) and quantified percentages (right) of Ki-67 + , IFNγ + , and CD38 + cells among CD4 + T cells ( d , f , h ) and CD8 + T cells ( e , g , i ) after 72 h of PBMCs coculture in different groups (pooled data from 6 healthy donors across three independent experiments).The data are presented as the means ± SEMs and were analyzed by one-way ANOVA with the Bonferroni correction for multiple comparisons ( b – i ). All P values are two-sided and reported as exact values unless <0.001. Source data are provided as a Source Data file.

Article Snippet: Immunostaining was carried out via the following antibodies: anti-human CD4 (ab133616, Abcam, dilution 1:500), anti-human CD8 (ZA-0508, ZSGB-BIO, dilution 1:100), anti-mouse CD4 (25229S, CST, dilution 1:100), and anti-mouse CD8 (A23305PM, ABclonal, dilution 1:300).

Techniques: Cell Culture, Control

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

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.

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:

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.

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:

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.

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:

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.

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: