Journal: Materials Today Bio

Article Title: A smart all-in-one strategy based on hybrid bacteria with targeting drug delivery and spatiotemporal drug release to boost the synergistic therapeutic efficacy against TNBC

doi: 10.1016/j.mtbio.2025.101849

Figure Lengend Snippet: Evaluation of antitumor immune responses and biosafety evaluation. (A) FCM analysis and quantitative analysis of (B) CD4 + T cells and (C) CD8 + T cells in tumor tissues. The levels of (D) IL-6, (E) IFN-γ and (F) TNF-α in the serum of mice quantified by ELISA at the end of treatment. (G) Body weight of mice after various treatments (n = 4). (H) H&E staining of the major organs (heart, liver, spleen, lung and kidney) of mice after various treatments (Scale bars: 100 μm). Data are presented as mean ± SD (ns, not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: Cell counting kit-8 (CCK-8), PE Anti-mouse CD3, FITC Anti-mouse CD8a and APC Anti-mouse CD4 were obtained from Elabscience Biotechnology Co., Ltd (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Frontiers in Immunology

Article Title: Immune regulatory adjuvant approach to mitigate subcutaneous immunogenicity of monoclonal antibodies

doi: 10.3389/fimmu.2024.1496169

Figure Lengend Snippet: Impact of OPLS on RTX- and ADM-mediated T-cell activation when administered subcutaneously in mice. CD4 + T cells in the draining lymph nodes (DLN) of mice at the study terminal were stained for the activation marker CD44 and classical immune checkpoints CLTA-4 and PD-1. Supplementary Figure 3 is the flow cytometry gating strategy. Frequency (%) of (A) CD44 ++ and (B) CTLA-4 + PD-1 + cells out of live CD3 + CD4 + T cells in DLN of mice administered RTX + rHuPH20 in vehicle, 25 mM OPLS, or 100 mM OPLS or (C) in DLN of mice administered ADM in vehicle, 25 mM OPLS, or 50 mM OPLS. Dots represent individual mice and bars are mean ± SD. Statistical significance was determined by Student’s unpaired t-test (one-tailed). *p<0.05, **p<0.01.

Article Snippet: Where indicated, splenocytes were surface stained with anti-mouse CD4 APC-Cy7 (GK1.5), CD3 FITC (17A2), LAG-3 APC (C9B7W), CD49 PE (HMA2), and LAP PerCP-Cy5.5 (TW7-16B4) and stained intracellularly with anti-mouse FoxP3 Alexa Fluor 700 (MF-14) using a fixation/permeabilization kit (Tonbo Biosciences).

Techniques: Activation Assay, Staining, Marker, Flow Cytometry, One-tailed Test

Journal: Frontiers in Immunology

Article Title: Immune regulatory adjuvant approach to mitigate subcutaneous immunogenicity of monoclonal antibodies

doi: 10.3389/fimmu.2024.1496169

Figure Lengend Snippet: Co-administration with OPLS promotes differentiation of regulatory T cells in vivo and in vitro . In the ADM study ( Figure 1 ), mouse splenocytes were collected at the terminal endpoint and analyzed for Tr1 phenotype. Supplementary Figure 4 is the flow cytometry gating strategy. (A) Frequency (%) of LAG-3 + CD49b + FoxP3 - CD4 + Tr1 cells in the spleen. Dots represent individual mice and bars are mean ± SD. Statistical significance was determined by Student’s unpaired t-test (one-tailed). *p<0.05. (B, C) Naïve immature SW mouse BMDCs were cultured for 24 h with media, VitD3/Dex, OVA (a model antigen), and OVA + 25 mM or 50 mM OPLS. Splenocytes from SW mice (n=2) subcutaneously immunized with OVA (2 μg/100 μL) once weekly were collected 4 days after the second dose. Isolated CD4 + T cells were co-cultured with treated BMDCs at a ratio of 1:5 BMDC, CD4 + T cell for 72 h. Frequency (%) (B) LAG-3 + CD49 + Tr1 and (C) LAP + of CD4 + FoxP3 - T cells. Error bars are mean ± SD of triplicate wells.

Article Snippet: Where indicated, splenocytes were surface stained with anti-mouse CD4 APC-Cy7 (GK1.5), CD3 FITC (17A2), LAG-3 APC (C9B7W), CD49 PE (HMA2), and LAP PerCP-Cy5.5 (TW7-16B4) and stained intracellularly with anti-mouse FoxP3 Alexa Fluor 700 (MF-14) using a fixation/permeabilization kit (Tonbo Biosciences).

Techniques: In Vivo, In Vitro, Flow Cytometry, One-tailed Test, Cell Culture, Isolation

Journal: Frontiers in Immunology

Article Title: Immune regulatory adjuvant approach to mitigate subcutaneous immunogenicity of monoclonal antibodies

doi: 10.3389/fimmu.2024.1496169

Figure Lengend Snippet: OPLS does not impact immunocompetence of NHP and mice. CD-1 mice received daily SC doses of vehicle (n=6), 20 mg/kg CYP (n=3), or OPLS (n=6/group) at 18-450 mM for 28 days. NHP (rhesus macaques) (n=3) received 21 daily SC doses of 54 mM OPLS. (A, B) Frequencies of lymphocyte populations—CD19 + B cells, CD3 + T cells, CD3 + CD4 + T cells, and CD3 + CD8 + T cells—in (A) spleens of mice on day 28 and (B) peripheral blood of NHP on day 0 (baseline), 8, 18, 22, and 35. Supplementary Figure 5 is the gating strategy. (C) Anti-KLH IgM and (D) IgG titers (log 2 ) in plasma of mice on day 28 following a single IV dose of KLH (2 mg) on day 15. (E) Anti-KLH IgM and IgG titers (log 2 ) in plasma of NHP on day 18 and 22, respectively, following a single IM dose of KLH (10 mg) on day 8. Each dot represents an individual animal and all bars are mean ± SD. (F-K) Microscopic images of H&E-stained spleens from mice treated with (F) vehicle or (G) 18 mM, (H) 45 mM, (I) 90 mM, (J) 225 mM, and (K) 450 mM OPLS. Statistical significance was determined by one-way ANOVA with Dunnett’s multiple comparisons test. ****p<0.0001.

Article Snippet: Where indicated, splenocytes were surface stained with anti-mouse CD4 APC-Cy7 (GK1.5), CD3 FITC (17A2), LAG-3 APC (C9B7W), CD49 PE (HMA2), and LAP PerCP-Cy5.5 (TW7-16B4) and stained intracellularly with anti-mouse FoxP3 Alexa Fluor 700 (MF-14) using a fixation/permeabilization kit (Tonbo Biosciences).

Techniques: Clinical Proteomics, Staining

Journal: Cell reports

Article Title: Temporal dynamics of immune cell transcriptomics in brain metastasis progression influenced by gut microbiome dysbiosis

doi: 10.1016/j.celrep.2025.115356

Figure Lengend Snippet:

Article Snippet: Flow: rat anti-CD4-APC-Cy7 (RM4–5) , Tonbo , Cat#25–0042-U025; RRID: N/A.

Techniques: In Vivo, Recombinant, Blocking Assay, Red Blood Cell Lysis, Multiplex Assay, Sequencing, Software, Amplification, Fluorescence

Journal: Current Microbiology

Article Title: Innate and Adaptive Immune Responses Induced by Aspergillus fumigatus Conidia and Hyphae

doi: 10.1007/s00284-022-03102-1

Figure Lengend Snippet: T cell expression of human PBMCs after stimulation by A. fumigatus conidia and hyphae. a CD4/CD8 ratio, b Th1, c Th2, d Th17. Each geometric symbol represents a sponsor ( n =4). Live Conidia LC, Live Hyphal fragments LH, Heat-inactivated Conidia HIC, Heat-inactivated Hyphal fragments HIH

Article Snippet: Next, cells were collected and washed with PBS, followed by extracellular staining with anti-human CD3 PE-Cy7, CD4 APC-Cy7 and CD8 PE-Cy5.5 for 15 min. After that, PBMCs were centrifuged and resuspended in Fixation/permeabilisation reagents (Biogems, USA) for 30 min to perform the subsequent intracellular cytokine staining with anti-human IL-4 R-PE, IL-17a APC and IFN-γ Alexa FlourTM 700 (all antibodies: BioLegend, USA).

Techniques: Expressing

Journal: Asian Journal of Pharmaceutical Sciences

Article Title: Lignin-assisted construction of sub-10 nm supramolecular self-assembly for photothermal immunotherapy and potentiating anti-PD-1 therapy against primary and distant breast tumors

doi: 10.1016/j.ajps.2022.07.002

Figure Lengend Snippet: In vivo immune responses assessment. (A) Flow cytometry assay of CD80 + or CD86 + gated on CD11c + in the TDLNs of mice. (B) Flow cytometry assay illustrated the percent of CD8a + T cells and CD4 + T cells gated on CD3 + T cells in splenocytes of mice. (C-D) Quantitative percentages of CD80 + or CD86 + expressions gated on CD11c + according to (A). (E) Quantitative percentages of CD8a + T cells and CD4 + T cells in splenocytes according to (B). (F-H) Quantitative analysis of the immune-associated cytokines in serum, as determined by ELISA (IL-2, TNF-α and IFN-γ). ( n = 3, mean ± SD, * P < 0.05, ** P < 0.01 and ## P < 0.0001).

Article Snippet: Fluorochrome-labeled anti-mouse monoclonal antibodies (CD11c-allophycocyanin (APC), CD80-fluorescein isothiocyanate (FITC), CD86-phycoerythrin (PE), CD3-FITC, CD8a-PE, and CD4-APC) were bought from Proteintech.

Techniques: In Vivo, Flow Cytometry, Enzyme-linked Immunosorbent Assay