Journal: Heliyon

Article Title: Erzhi pills reverse PD-L1-mediated immunosuppression in melanoma microenvironment

doi: 10.1016/j.heliyon.2024.e24988

Figure Lengend Snippet: EZP increased the percentage of CD4 + T cells and enhanced the function of CD8 + T in vivo . (A–D) T cells were analyzed by flow cytometry. (A) The gate strategy for CD4 + and CD8 + T cells. Flow cytometry plots (B) and bar graphs present percentages of CD3 + CD4 + T cells (C) and CD3 + CD8 + T cells (D) in the spleen from each group mice. n = 5. (E–H) IHC staining for the tumor tissues. IHC images (E) and quantifications of CD4 + T cells (F), CD8 + T cells (G) and GZMB + cells (H) in tumor tissues. Scale bar, 100 μm. n = 3. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01 and *** P < 0.001 vs DMSO. IHC, immunohistochemistry; GZMB, granzyme B.

Article Snippet: PerCP/Cyanine5.5 anti-mouse CD3 antibody (E-AB-F1013J), FITC anti-mouse CD8a antibody (E-AB-F1104C) and PE anti-mouse CD4 antibody (E-AB-F1097D) were obtained from Elabscience (Wuhan, China).

Techniques: In Vivo, Flow Cytometry, Immunohistochemistry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: XFab-α4-1BB/CD40L fusion protein activates dendritic cells, improves expansion of antigen-specific T cells, and exhibits antitumour efficacy in multiple solid tumour models

doi: 10.1007/s00262-023-03535-y

Figure Lengend Snippet: XFab-α4-1BB/CD40L fusion protein enhances T-cell activation and proliferation in vitro. CellTrace Violet-labelled human T cells were cocultured with ovalbumin–loaded allogenic dendritic cells (DCs) in the presence of titrated bispecific antibody or control antibodies for 10 days. Proliferation (CellTrace Violet dilution) and CD25 and HLA-DR expression of CD4+ or CD8+ cells were analysed by flow cytometry. The percentage of T-cell proliferation (a) and the percentage of CD25 and HLA-DR expression (b) are shown as mean ± SD (n = technical duplicates). Dotted lines indicate cultures in the absence of treatment. c XFab-α4-1BB/CD40L reinforces the immunological synapse through target crosslinking. Representative fluorescent images of DC-T cocultures were used to co-localise FITC-CD3, PE-XFab-α4-1BB/CD40L, and APC-CD8. The white dashed line represents the interface between the DC and T cells. Images were acquired at 40×magnification. Scale bars: 20 μm

Article Snippet: Other antibodies include H-2 Kb OVA Tetramer-SIINFEKL (TS-5001-1C, MBL International), anti-mouse CD8 FITC (D271-4, MBL International anti-mouse), and anti-human Foxp3 PE (320008, Invitrogen).

Techniques: Activation Assay, In Vitro, Expressing, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: XFab-α4-1BB/CD40L fusion protein activates dendritic cells, improves expansion of antigen-specific T cells, and exhibits antitumour efficacy in multiple solid tumour models

doi: 10.1007/s00262-023-03535-y

Figure Lengend Snippet: Repeated dosing of XFab-α4-1BB/CD40L in OVA-rechallenged mice results in expansion of OVA-specific CD8+ cells and inhibited tumour growth in B16-OVA-bearing mice. Human CD40/4-1BB transgenic mice were immunised with 12.5 mg/kg OVA protein and administered indicated dosage of therapeutic agents or 1.25 mg/kg poly (I:C) intradermally on five occasions. On day 24, 5 × 105 B16-OVA cells were inoculated subcutaneously. Whole blood and tumour tissues were collected for flow cytometry. a Overview of the experimental set-up. b Tumour growth after B16-OVA inoculation. Tumour is displayed until the point where the first mouse in each group approached the ethical tumour volume limit of 2 cm3 and is sacrificed. c Total counts of OVA-specific CTL cells (CD3+ CD8+ CD44+ H-2Kb -OVA tetramer+, left) and total counts of CD8+ effector memory (EM) T cells (CD3+ CD8+ CD44+ CD62L−, right) in blood. d Total counts of OVA-specific CTL cells (left) and total counts of CD8+ EM T cells (right) in tumour tissues on day 50. Data points indicate mean ± SEM. n = 6–7 per group

Article Snippet: Other antibodies include H-2 Kb OVA Tetramer-SIINFEKL (TS-5001-1C, MBL International), anti-mouse CD8 FITC (D271-4, MBL International anti-mouse), and anti-human Foxp3 PE (320008, Invitrogen).

Techniques: Transgenic Assay, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: XFab-α4-1BB/CD40L fusion protein activates dendritic cells, improves expansion of antigen-specific T cells, and exhibits antitumour efficacy in multiple solid tumour models

doi: 10.1007/s00262-023-03535-y

Figure Lengend Snippet: XFab-α4-1BB/CD40L inhibits tumour growth and increases intratumoural T-cell accumulation and activation in MC38-bearing mice. a Randomised human CD40/4-1BB transgenic mice were subcutaneously implanted with MC38 cells at 2.0 × 105 per mouse. When tumours were approximately 50–90 mm3, the mice were intratumourally administered designated antibodies or vehicle at indicated time points. Tumour burden was quantitated at various time points after treatment. Data points indicate mean ± SEM (n = 8 animals per group). Statistical analysis was performed by two-way repeated measure ANOVA with Tukey’s multiple comparison test. b Kaplan–Meier survival curves of MC38-bearing mice. Statistical significance was analysed by the log-rank test. c Digested tumour tissues at the end point were analysed by flow cytometry gating on living CD4+ or CD8+ cells. Shown is mean ± SD. Tumour-infiltrating CD4+ and CD8+ T cells were analysed for memory cell subpopulation (d) and 4-1BB and PD-1 expression (e) by flow cytometry. Effector memory (EM) and central memory (CM) T cells were defined as CD44+CD62L− and CD44+CD62L+ T cells, respectively. Data points indicate mean ± SD (n = 8 animals per group). Statistical significance in (c–e) was calculated using unpaired one-way ANOVA with Tukey’s multiple comparison test

Article Snippet: Other antibodies include H-2 Kb OVA Tetramer-SIINFEKL (TS-5001-1C, MBL International), anti-mouse CD8 FITC (D271-4, MBL International anti-mouse), and anti-human Foxp3 PE (320008, Invitrogen).

Techniques: Activation Assay, Transgenic Assay, Comparison, Flow Cytometry, Expressing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: XFab-α4-1BB/CD40L fusion protein activates dendritic cells, improves expansion of antigen-specific T cells, and exhibits antitumour efficacy in multiple solid tumour models

doi: 10.1007/s00262-023-03535-y

Figure Lengend Snippet: Effect of XFab-α4-1BB/CD40L in combination with gemcitabine on the inhibition of tumour growth in Panc02 cell-bearing mice. a Human CD40 × 4-1BB transgenic mice were subcutaneously implanted with Panc02 cells at 5.0 × 106 per mouse. When tumours were approximately 50–90 mm3, the mice were administered 30 mg/kg gemcitabine intravenously only or in combination with 50 nmol/kg XFab-α4-1BB/CD40L intratumourally at the indicated time points. Statistical analysis was performed by two-way repeated measure ANOVA with Tukey’s multiple comparison test. b Digested tumour tissues at the end point were analysed by flow cytometry gating on Treg and CD4 cells. c Tumour-infiltrating CD4+ and CD8+ T cells were analysed for 4-1BB and PD-1 expression by flow cytometry. Data points in (b, c) indicate mean ± SD (n = 6–8 animals per group), and statistical significance was calculated using unpaired one-way ANOVA with Tukey’s multiple comparison test

Article Snippet: Other antibodies include H-2 Kb OVA Tetramer-SIINFEKL (TS-5001-1C, MBL International), anti-mouse CD8 FITC (D271-4, MBL International anti-mouse), and anti-human Foxp3 PE (320008, Invitrogen).

Techniques: Inhibition, Transgenic Assay, Comparison, Flow Cytometry, Expressing