Journal: Drug Delivery

Article Title: Lipid nanoparticles with aptamers enable targeted mRNA delivery to CD4⁺ T cells

doi: 10.1080/10717544.2026.2637266

Figure Lengend Snippet: (A) Schematic representation of aptamer–lipid nanoparticle (LNP) formulation using a microfluidic device, followed by thiol–aptamer conjugation. (B) Hydrodynamic size and (C) polydispersity index (PDI) of MC3- and SM102-based LNPs, either non-targeting or Apt62-modified. (D) Schematic illustrating molecular beacon hybridization assay for quantifying aptamer density on LNPs. (E) Thermal melt curves of Apt62 molecular beacons at concentrations ranging from 0 to 800 nM. (F) Biolayer interferometry (BLI) sensograms of Apt62 binding to recombinant human CD4 protein. (G) BLI sensograms of Apt62-functionalized LNPs binding to recombinant human CD4 protein. Data represent mean±SD ( n = 3 for B–C).

Article Snippet: Fetal Bovine Serum (FBS, Cat. No. A31605-01), LIVE/DEADTM Fixable Far Red Dead Cell Stain Kit (Cat. No. L10120 ), anti-human CD3-PE (Cat. No. 12-0036-42) monoclonal antibody, human CD4 monoclonal antibody (Cat. No. 69-0049-42), mouse CD4 monoclonal antibody (Cat. No. 14-0042-82), d-Luciferin (Cat. No. L2916), antibiotic-antimycotic (Cat. No. 15240062), Tris-EDTA buffer (TE, 20X, Cat. No. P7589), NaCl (Cat. No. AM9759), and MgCl 2 (Cat. No. AM9530G), were purchased from ThermoFisher Scientific (Waltham, MA).

Techniques: Formulation, Conjugation Assay, Modification, Hybridization, Binding Assay, Recombinant

Journal: Drug Delivery

Article Title: Lipid nanoparticles with aptamers enable targeted mRNA delivery to CD4⁺ T cells

doi: 10.1080/10717544.2026.2637266

Figure Lengend Snippet: (A) eGFP expression in SUP-T1 (CD4⁺) and HSB2 (CD4⁻) cells following treatment with mRNA-loaded LNPs formulated with MC3 or SM102, either non-targeting, Apt62-modified (25:1 or 100:1), or anti-human CD4 mAb-conjugated. (B) Representative flow cytometry histograms showing eGFP signal in live, singlet-gated in SUP-T1 (CD4⁺) and HSB2 (CD4⁻) cells that were untreated (non-transfected) or treated with SM102-Apt62:100 or SM102 anti-human CD4 mAb-modified LNPs. (C) Quantification of mean fluorescence intensity (MFI) of SUP-T1 (CD4⁺) and HSB2 (CD4⁻) populations corresponding to panel (B). Data represent mean±SD ( n = 4). Statistical significance was determined using two-way ANOVA followed by Tukey's multiple comparisons test; **** p < 0.0001, *** p < 0.001, ** p < 0.01, ####p < 0.0001 versus corresponding CD4⁺ population.

Article Snippet: Fetal Bovine Serum (FBS, Cat. No. A31605-01), LIVE/DEADTM Fixable Far Red Dead Cell Stain Kit (Cat. No. L10120 ), anti-human CD3-PE (Cat. No. 12-0036-42) monoclonal antibody, human CD4 monoclonal antibody (Cat. No. 69-0049-42), mouse CD4 monoclonal antibody (Cat. No. 14-0042-82), d-Luciferin (Cat. No. L2916), antibiotic-antimycotic (Cat. No. 15240062), Tris-EDTA buffer (TE, 20X, Cat. No. P7589), NaCl (Cat. No. AM9759), and MgCl 2 (Cat. No. AM9530G), were purchased from ThermoFisher Scientific (Waltham, MA).

Techniques: Expressing, Modification, Flow Cytometry, Transfection, Fluorescence

Journal: Drug Delivery

Article Title: Lipid nanoparticles with aptamers enable targeted mRNA delivery to CD4⁺ T cells

doi: 10.1080/10717544.2026.2637266

Figure Lengend Snippet: (A) Representative ex vivo IVIS images of major organs collected 6 h post-injection from mice treated with saline, MC3 or SM102 LNPs (non-targeting or Apt62:75), or SM102 anti-mouse/human CD4 mAb-conjugated LNPs. (B) Quantification of normalized luminescence in the liver. (C) Quantification of normalized luminescence in spleen. (D) Percentage of spleen-to-liver luminescence ratio. Data represent mean±SD ( n = 4). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test; **** p < 0.0001, *** p < 0.001, ** p < 0.01.

Article Snippet: Fetal Bovine Serum (FBS, Cat. No. A31605-01), LIVE/DEADTM Fixable Far Red Dead Cell Stain Kit (Cat. No. L10120 ), anti-human CD3-PE (Cat. No. 12-0036-42) monoclonal antibody, human CD4 monoclonal antibody (Cat. No. 69-0049-42), mouse CD4 monoclonal antibody (Cat. No. 14-0042-82), d-Luciferin (Cat. No. L2916), antibiotic-antimycotic (Cat. No. 15240062), Tris-EDTA buffer (TE, 20X, Cat. No. P7589), NaCl (Cat. No. AM9759), and MgCl 2 (Cat. No. AM9530G), were purchased from ThermoFisher Scientific (Waltham, MA).

Techniques: Ex Vivo, Injection, Saline

Journal: Drug Delivery

Article Title: Lipid nanoparticles with aptamers enable targeted mRNA delivery to CD4⁺ T cells

doi: 10.1080/10717544.2026.2637266

Figure Lengend Snippet: Assessment of organ indices and systemic cytokine markers 6 h postinjection. (A) Liver-to-body weight ratio. (B) Spleen-to-body weight ratio. (C) Serum IL-6 levels. (D) Serum TNF- α levels. (E) Serum ALT levels. The mice were treated with saline, MC3 or SM102 LNPs (nontargeting or Apt62:75), or SM102 anti-mouse CD4 mAb-conjugated LNPs. Data represent the mean±SD ( n = 4). Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01.

Article Snippet: Fetal Bovine Serum (FBS, Cat. No. A31605-01), LIVE/DEADTM Fixable Far Red Dead Cell Stain Kit (Cat. No. L10120 ), anti-human CD3-PE (Cat. No. 12-0036-42) monoclonal antibody, human CD4 monoclonal antibody (Cat. No. 69-0049-42), mouse CD4 monoclonal antibody (Cat. No. 14-0042-82), d-Luciferin (Cat. No. L2916), antibiotic-antimycotic (Cat. No. 15240062), Tris-EDTA buffer (TE, 20X, Cat. No. P7589), NaCl (Cat. No. AM9759), and MgCl 2 (Cat. No. AM9530G), were purchased from ThermoFisher Scientific (Waltham, MA).

Techniques: Saline

Journal: Drug Delivery

Article Title: Lipid nanoparticles with aptamers enable targeted mRNA delivery to CD4⁺ T cells

doi: 10.1080/10717544.2026.2637266

Figure Lengend Snippet: eGFP expression in SUP-T1 (CD4⁺) and HSB2 (CD4⁻) cells following transfection with SM102 LNPs functionalized with AutoNA-predicted aptamers (Apt62, Apt54, Apt72, Apt121) at a 100:1 ratio. Data represent the mean±SD ( n = 4). Statistical significance was determined using two-way ANOVA followed by Tukey’s multiple comparisons test; **** p < 0.0001.

Article Snippet: Fetal Bovine Serum (FBS, Cat. No. A31605-01), LIVE/DEADTM Fixable Far Red Dead Cell Stain Kit (Cat. No. L10120 ), anti-human CD3-PE (Cat. No. 12-0036-42) monoclonal antibody, human CD4 monoclonal antibody (Cat. No. 69-0049-42), mouse CD4 monoclonal antibody (Cat. No. 14-0042-82), d-Luciferin (Cat. No. L2916), antibiotic-antimycotic (Cat. No. 15240062), Tris-EDTA buffer (TE, 20X, Cat. No. P7589), NaCl (Cat. No. AM9759), and MgCl 2 (Cat. No. AM9530G), were purchased from ThermoFisher Scientific (Waltham, MA).

Techniques: Expressing, Transfection

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: Frontiers in Immunology

Article Title: Peripheral blood T-cell subsets combined with EGRIS score predict the need for mechanical ventilation in Guillain–Barré syndrome

doi: 10.3389/fimmu.2026.1747416

Figure Lengend Snippet: Association of peripheral blood lymphocyte counts with the need for mechanical ventilation in patients with Guillain–Barré syndrome (GBS). (A) ROC analysis of CD8 + T cell and CD4 + T cell counts and the predictive value of Erasmus GBS Respiratory Insufficiency Score (EGRIS) for mechanical ventilation in GBS. P < 0.05 was considered significant. (B) Multivariate logistic regression analysis of gender, age, CD8 + T cell counts and CD4 + T cell counts and EGRIS in GBS patients requiring mechanical ventilation. P < 0.05 was considered significant.

Article Snippet: Sections were incubated with mouse anti-CD4 monoclonal antibody (Protein tech) overnight at 4°C and then with FITC-conjugated goat anti-mouse IgG (H + L) (Boster Biological Technology) for 1 hour at room temperature.

Techniques:

Journal: Frontiers in Immunology

Article Title: Peripheral blood T-cell subsets combined with EGRIS score predict the need for mechanical ventilation in Guillain–Barré syndrome

doi: 10.3389/fimmu.2026.1747416

Figure Lengend Snippet: CD4 + T cell expression in the sciatic nerve of experimental autoimmune neuritis (EAN) rats, n = 4. (A) Immunofluorescence staining of rat sciatic nerve tissue. (original magnification, 40×). Green fluorescence indicates CD4 + T cells, blue fluorescence indicates DAPI, and the cells outlined in white and green represent CD4 + T cells. (B) Preparation of EAN rats, neurological function monitoring, and tissue collection timeline. (C) Neurological function score curve of EAN rats within 30 days after immunization. (D) Cell counts are expressed as mean ± standard deviation (SD). One-way ANOVA was used to assess the number of CD4 + T cells in the sciatic nerves of EAN rats at different disease stages, followed by Tukey’s post hoc test for multiple group comparisons. A p-value < 0.05 was considered statistically significant. Statistical differences between groups are indicated by asterisks: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Sections were incubated with mouse anti-CD4 monoclonal antibody (Protein tech) overnight at 4°C and then with FITC-conjugated goat anti-mouse IgG (H + L) (Boster Biological Technology) for 1 hour at room temperature.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Standard Deviation

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:

Journal: NPJ Vaccines

Article Title: Lipid nanoparticle encapsulated TLR3 agonist adjuvant elicits potent T cell immunity against cancer and viruses

doi: 10.1038/s41541-025-01349-w

Figure Lengend Snippet: a C57BL/6 mice were immunized intramuscularly with 10 μg of GP33 and 10 μg of GP66 formulated with 10 μg of NVT, NVT/DOTAP-LNP, NVT/SM-LNP, or equivalent amounts of empty DOTAP-LNP or empty SM-LNP on days 7, 14, and 21. On day 28, GP33-specific CD8 + T cell responses and GP66-specific CD4 + T cell responses in PBMCs were measured by flow cytometry. n = 5. b C57BL/6 mice were immunized intramuscularly with 10 µg of GP33 formulated with 1, 3, or 10 µg of NVT/LNP, and GP33-specific CD8 + T cell responses in PBMCs were measured by flow cytometry. n = 5. c Body weight of C57BL/6 mice was monitored from day 0 to day 4 following the first intramuscular immunization with GP33 formulated with 1, 3, or 10 µg of NVT/LNP as described in ( b ). Body weight is expressed as a percentage of the baseline weight on day 0. Data are presented as median (min–max). n = 5. d C57BL/6 mice were immunized intramuscularly with 1, 3, 10, or 30 µg of GP33 formulated with 10 µg of NVT/LNP, and GP33-specific CD8 + T cell responses in PBMCs were measured by flow cytometry. n = 5. The data are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparisons test. * P < 0.05; **** P < 0.0001.

Article Snippet: For T cell depletion, InVivo MAb anti-CD4 monoclonal antibody (clone GK1.5, BE0003-1, BioXCell, Lebanon, NH, USA) and InVivo MAb anti-CD8α monoclonal antibody (clone 2.43, BE0061, BioXCell) were administered intraperitoneally.

Techniques: Flow Cytometry

Journal: NPJ Vaccines

Article Title: Lipid nanoparticle encapsulated TLR3 agonist adjuvant elicits potent T cell immunity against cancer and viruses

doi: 10.1038/s41541-025-01349-w

Figure Lengend Snippet: Mice were intramuscularly vaccinated with 10 μg of NVT/LNP-formulated peptide vaccines (10 μg of peptide antigen) on days 7, 14, and 21 in tumor models. Tumor progression and survival were monitored over time. a C57BL/6 mice were subcutaneously injected with 5 × 10 5 TC-1-Luc cells and treated with NVT/LNP containing or lacking the E7 peptide. On day 24, E7-specific CD8 + T cell responses were assessed from isolated PBMCs. n = 4, 5, 6. This experiment was independently repeated three times, and data from one representative experiment are shown. b In the cervical cancer model, mice were intravaginally challenged with 5 × 10 5 TC-1-Luc cells and immunized with E7 peptide-formulated NVT/LNP, n = 6, or PBS control, n = 5. Representative bioluminescence images of mice were captured using a VISQUE® Smart-LF imaging system. c For the lung metastasis model, mice were intravenously injected with 5 × 10 5 TC-1-Luc cells and vaccinated as in ( b ). E7-specific CD8 + T cell responses were analyzed from surviving mice on day 83, followed by tumor rechallenge on day 90 via tail vein injection. n = 6 (1 st challenge); 10, 4 (2 nd challenge). Representative bioluminescence images of mice were captured using a VISQUE® Smart-LF imaging system. This experiment was independently repeated twice, and data from one representative experiment are shown. d Tumor-bearing mice were treated with NVT/LNP vaccines intramuscularly, and anti-CD4 or anti-CD8 antibodies (200 µg) were administered intraperitoneally twice weekly from day 7, for a total of six doses. n = 6. e C57BL/6 mice were subcutaneously inoculated with 5 × 10 5 B16F10-OVA cells and vaccinated with OVA CD8 peptide-formulated NVT/LNP. Tumor burden at day 18 is indicated by a red dotted line, and OVA-specific CD8 + T cells were quantified on day 22 by tetramer staining of PBMCs. Representative images of mice were captured using a digital camera prior to tumor measuring on day 18. n = 5. This experiment was independently repeated three times, and data from one representative experiment are shown. f Following intravenous injection of 5 × 10 5 B16F10-OVA cells, mice were vaccinated with OVA peptide-formulated NVT/LNP. Lung tumor burden was determined on day 30 by counting tumor nodules after sacrifice. Representative images of lung were captured using a digital camera prior to nodule counting. n = 4, 5. This experiment was independently repeated twice, and data from one representative experiment are shown. g C57BL/6 mice were infected with LCMV and immunized intramuscularly with GP33 peptide-formulated NVT/LNP at 14-, 21-, and 28-days post-infection. Viral titers in the blood were measured on day 40. n = 5. Data are shown as mean ± SEM for tumor growth, and mean ± SD for T cell responses and viral titers. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test. ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s. not significant.

Article Snippet: For T cell depletion, InVivo MAb anti-CD4 monoclonal antibody (clone GK1.5, BE0003-1, BioXCell, Lebanon, NH, USA) and InVivo MAb anti-CD8α monoclonal antibody (clone 2.43, BE0061, BioXCell) were administered intraperitoneally.

Techniques: Vaccines, Injection, Isolation, Control, Imaging, Staining, Infection

Journal: NPJ Vaccines

Article Title: Lipid nanoparticle encapsulated TLR3 agonist adjuvant elicits potent T cell immunity against cancer and viruses

doi: 10.1038/s41541-025-01349-w

Figure Lengend Snippet: a C57BL/6 mice were immunized intramuscularly on days 7, 14, and 21 with 10 μg each of GP33 and GP66 antigens, formulated with 10 μg of CpG, NVT, poly(I:C), GM-CSF, or with 10 μg of CpG, poly(I:C), or GM-CSF emulsified in IFA at a 1:1 (v/v) ratio with the antigens. Additional groups were administered 10 μg each of GP33 and GP66, formulated with 10 μg of CpG/LNP or NVT/LNP. On day 28, GP33-specific CD8 + T cell responses and GP66-specific CD4 + T cell responses in PBMCs were measured by flow cytometry. n = 5. b C57BL/6 mice were subcutaneously inoculated with 5 × 10 5 B16F10-OVA cells, and each adjuvant was administered intramuscularly in combination with the OVA CD8 peptide on days 7, 14, and 21 post-tumor implantations, vaccinated as in ( a ). Tumor growth was monitored over time. n = 5. c C57BL/6 mice were subcutaneously inoculated with 5 × 10 5 B16F10-OVA cells and vaccinated intramuscularly with OVA mRNA/SM-LNP or OVA peptide/NVT/SM-LNP vaccines on days 9, 16, and 23. For OVA mRNA antigen, doses of 1 or 5 µg were used, and for OVA peptide antigen, doses of 10 or 50 µg were applied. Tumor growth was monitored over time. On day 26, OVA-specific CD8 + T cell responses were measured from PBMCs. n = 5. d C57BL/6 mice were immunized intramuscularly with 10 μg each of TRP2 antigens, formulated with 10 μg of NVT/LNP or 10 μg of poly(I:C) emulsified in IFA at a 1:1 (v/v) ratio with the antigens on days 7, 14, and 21. Additional groups were administered 10 μg of CRM197-conjugated TRP2 (CRM-TRP2), formulated with 10 μg of poly(I:C) emulsified in IFA at a 1:1 (v/v) ratio with the antigens. On day 28, TRP2-specific CD8 + T cell responses in isolated PBMCs were measured by flow cytometry. n = 5. e C57BL/6 mice were subcutaneously inoculated with 5 × 10⁵ TC-1-Luc cells on day 0. Mice were then vaccinated with E7 peptide-NVT/LNP vaccines (1 µg peptide + 1 µg NVT/LNP) either intramuscularly or intratumorally on days 7, 14, and 21 post-tumor inoculation. Tumor growth was monitored over time. On day 21, peripheral blood mononuclear cells (PBMCs) were collected to measure E7-specific CD8⁺ T cell responses. n = 4. Data are shown as mean ± SEM for tumor growth, and mean ± SD for T cell responses and viral titers. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s., not significant.

Article Snippet: For T cell depletion, InVivo MAb anti-CD4 monoclonal antibody (clone GK1.5, BE0003-1, BioXCell, Lebanon, NH, USA) and InVivo MAb anti-CD8α monoclonal antibody (clone 2.43, BE0061, BioXCell) were administered intraperitoneally.

Techniques: Flow Cytometry, Adjuvant, Vaccines, Isolation