Journal: Frontiers in Oncology

Article Title: Bifidobacterium infantis-mediated HSV-TK/GCV therapy modulates the tumor microenvironment through site-specific phosphorylation of HIF-1α, mTOR, and PD-L1

doi: 10.3389/fonc.2026.1849164

Figure Lengend Snippet: Immunohistochemical validation in multiple mouse solid tumor models. (A–C) Tumor volume, weight, and growth curves (eight doses). (D) Kaplan-Meier survival curves. (E, F) IHC quantification of phospho-HIF-1α, phospho-PKM2, phospho-mTOR, and phospho-PD-L1 in immune-deficient (nude) mouse xenografts (n=6). (G, H) IHC quantification of the same phosphoproteins plus CD80 and CD86 in immune-competent (C57BL/6J) MC38 syngeneic tumors (n=5). *Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 (one-way ANOVA)*.

Article Snippet: The following is the antibody catalog: Phospho-PKM2-Y105 Rabbit pAb (Abclonal, Wuhan, China, AP0924), phospho-mTOR-Ser2448 (Abmart, Shanghai, China, T56571 ), phospho-HIF1A-Ser641/Ser643 (Affinity, Jiangsu, China, AF0062), CD80 Rabbit pAb (Abclonal, Wuhan, China, A16039), CD86 antibody (Abmart, Shanghai, China, T55238 ), Phospho-PD-L1/CD274-S283 polyclonal antibody (Biorbyt, Cambridge, United Kingdom, orb1722637).

Techniques: Immunohistochemical staining, Biomarker Discovery

Journal: Journal of Nanobiotechnology

Article Title: A multi species autophagy-inducing nano-vaccine platform: uncovering immunoenhancement properties of PCV2 cap nanoparticles

doi: 10.1186/s12951-026-04527-7

Figure Lengend Snippet: Inhibition of autophagy or TNF-α signaling reduced antigen presentation-related molecules. ( A to D ) 3D4 cells or BM-DCs were internalized with Cap NPs or ΔCap (2 µM) for 12 h, and the transcription levels of CD80 and CD86 were detected by qRT-PCR. BM-DCs were internalized with Cap NPs for 12 h, ( E ) Proportion analysis of CD80 + CD86 + double-positive cells in the CD11c + population of BM-DCs treated with Cap NPs. ( F and G ) Proportion analysis of CD80 + or CD86 + positive cells in the CD11c + population of BM-DCs treated with Cap NPs. 3D4 cells or BMDCs were incubated with Cap NPs and then cultured in fresh medium containing CQ for 12 h. The transcription of CD80 ( H and J ) and CD86 ( I and K ) was analyzed by qRT-PCR. 3D4 cells were transfected with si-ATG5 or negative control siRNA (si-NC) for 24 h, and then internalized with Cap NPs (2 µM) for 12 h. qRT-PCR was performed to test the transcription levels of CD80 ( L ) and CD86 ( M ). 3D4 or BMDC cells were incubated with Cap NPs and then cultured in fresh medium containing R7050 for 12 h. The transcription of CD80 ( N and P ) and CD86 ( O and Q ) was analyzed by qRT-PCR. One-way ANOVA was used for data analysis (** P < 0.01)

Article Snippet: To assess the protein expression levels of CD80 and CD86, BM-DCs internalized with Cap NPs were harvested and stained with a mixture of PE anti-mouse CD11c mAb, Alexa Fluor 488-conjugated rabbit anti-mouse CD80 mAb, and APC-conjugated rabbit anti-mouse CD86 mAb (ABclonal Technology, A27691 , A23965 and A25824 ) at the recommended concentrations for 30 min on ice.

Techniques: Inhibition, Immunopeptidomics, Quantitative RT-PCR, Incubation, Cell Culture, Transfection, Negative Control

Journal: Journal of Nanobiotechnology

Article Title: A multi species autophagy-inducing nano-vaccine platform: uncovering immunoenhancement properties of PCV2 cap nanoparticles

doi: 10.1186/s12951-026-04527-7

Figure Lengend Snippet: Cap peptide (193-213aa) activated autophagy and TNF-α related immune responses. ( A ) The schematic diagram of recombinant plasmids construction for full-length EGFP-Cap and its truncated forms. ( B ) 3D4 cells were first transfected with different EGFP-Cap plasmids or control plasmid (EGFP) for 24 h and then treated with rapamycin for 12 h. The transcription levels of TNF-α were detected by qRT-PCR. ( C ) The schematic diagram of mi3-Cap fragment recombinant plasmids construction. ( D ) Analysis of purified mi3, mi3-Cap fragment (mC-X) and Cap proteins in SDS-PAGE. ( E ) The self-assembly of mC-7-8, mC-8-9, and mC-9-10 NPs was presented by TEM. ( F ) 3D4 cells were internalized with different mi3 NPs for 12 h, and then cells were lysed. Western blotting was performed to evaluate the protein levels of p62, LC3 and His. β-actin was used as the loading control. ( G ) Ratios of LC3-II to β-actin of the panel F. ( H ) Ratios of p62 to β-actin of the panel F. ( I ) Stimulatory effects on multiple immune markers of mC-7-8, mC-8-9 and mC-9-10 NPs. qRT-PCR was used to detect the relative expression of TNF-α, IL-8, CD80, CD86, MHC-II, CXCL10, CCL20 and CSF1. Statistical significance was analyzed by one-way ANOVA (* P < 0.05; ** P < 0.01; ns P > 0.05)

Article Snippet: To assess the protein expression levels of CD80 and CD86, BM-DCs internalized with Cap NPs were harvested and stained with a mixture of PE anti-mouse CD11c mAb, Alexa Fluor 488-conjugated rabbit anti-mouse CD80 mAb, and APC-conjugated rabbit anti-mouse CD86 mAb (ABclonal Technology, A27691 , A23965 and A25824 ) at the recommended concentrations for 30 min on ice.

Techniques: Recombinant, Transfection, Control, Plasmid Preparation, Quantitative RT-PCR, Purification, SDS Page, Western Blot, Expressing

Journal: Journal of Nanobiotechnology

Article Title: A multi species autophagy-inducing nano-vaccine platform: uncovering immunoenhancement properties of PCV2 cap nanoparticles

doi: 10.1186/s12951-026-04527-7

Figure Lengend Snippet: A schematic illustration of PCV2 Cap NPs exploiting the autophagy-TNF-α pathway to enhance immunogenicity and their expanded application in the development of nano-vaccines. ( A ) The PCV2 Cap protein could self-assemble into icosahedral Cap NPs, enabling internalization into cells, while the mutant ΔCap was unable to complete NP assembly, impeding cell internalization. Subsequently, internalized Cap NPs induced the CaMKKβ-AMPK-mTOR pathway and elevated intracellular ROS levels, leading to autophagy activation. The increased level of autophagy facilitated the main processing and degradation of Cap NPs, while autophagy-deficient Cap NPs (ΔA-Cap NP) and autophagy-ubiquitin double deficient Cap NPs (ΔAU-Cap NP) decelerated their self-degradation. Moreover, activated autophagy elevated the immunoregulatory signal TNF-α and antigen presentation-related molecules CD80, CD86, and MHC-II. The autophagy-TNF-α signaling induced by Cap NPs promoted humoral and cellular immunity in mice. ( B ) In this study, a functional Cap peptide (193-213aa) was identified based on the immune enhancing mechanisms of Cap NPs. Compared with mi3 NPs, the chimeric mi3-Cap peptide NPs could trigger the autophagy-TNF-α pathway and stimulate the expression of a series of immune activation markers, suggesting that the functional peptide may serve as a novel immune adjuvant. Furthermore, Cap NPs were also found to internalize and activate the autophagy-TNF-α pathway in cells of multiple species, making them candidates for a broad-spectrum antigen delivery nano-vaccine platform. These findings hold promise for developing novel vaccines with higher immunogenicity in the future

Article Snippet: To assess the protein expression levels of CD80 and CD86, BM-DCs internalized with Cap NPs were harvested and stained with a mixture of PE anti-mouse CD11c mAb, Alexa Fluor 488-conjugated rabbit anti-mouse CD80 mAb, and APC-conjugated rabbit anti-mouse CD86 mAb (ABclonal Technology, A27691 , A23965 and A25824 ) at the recommended concentrations for 30 min on ice.

Techniques: Immunopeptidomics, Vaccines, Mutagenesis, Activation Assay, Ubiquitin Proteomics, Functional Assay, Expressing, Adjuvant

Journal: Cancer Science

Article Title: Triple‐Mutated HSV ‐1 Expressing Soluble B7 ‐1 Plus CTLA ‐4 Blockade Suppresses Lymph Node Metastasis in Tongue Cancer

doi: 10.1111/cas.70410

Figure Lengend Snippet: Genome structures of T‐01 and T‐mB7‐1. Both T‐01 and T‐mB7‐1 are based on the G47Δ backbone, which contains triple deletions in both copies of the γ34.5 gene, as well as in the ICP6 and α47 genes. T‐mB7‐1 additionally carries a cDNA encoding the extracellular domain (ECD) of murine B7‐1 fused to the Fc region of human IgG1, inserted into the ICP6 locus under the control of the immediate‐early CMV promoter. T‐01 serves as a control virus, containing an empty cassette inserted at the same locus. Abbreviations: TR L , terminal repeat long; U L , unique long; IR L , inverted repeat long; IR S , inverted repeat short; U S , unique short; TR S , terminal repeat short; ICP6, infected cell protein 6; lacZ, Escherichia coli LacZ; pA, poly A; p‐CMV, immediate‐early cytomegalovirus promoter.

Article Snippet: Vero, SCCVII, and SAS cells were seeded in six‐well plates at a density of 2.0 × 10 5 cells per well and incubated for 18 h. The cells were then infected with viruses at a multiplicity of infection of 1.0 for 1 h. After overnight incubation at 34.5°C, the concentration of soluble B7‐1 in the culture supernatant was determined using a mouse CD80 ELISA kit (RayBiotech, GA, USA).

Techniques: Control, Virus, Infection

Journal: Cancer Science

Article Title: Triple‐Mutated HSV ‐1 Expressing Soluble B7 ‐1 Plus CTLA ‐4 Blockade Suppresses Lymph Node Metastasis in Tongue Cancer

doi: 10.1111/cas.70410

Figure Lengend Snippet: Expression of soluble B7‐1 and cytopathic activity of T‐mB7‐1 in human and murine squamous cell carcinoma cell lines. (a) Concentrations of soluble B7‐1 secreted into the culture supernatant following infection of Vero, SAS (human OSCC), and SCCVII (murine OSCC) cells with T‐mB7‐1 or T‐01, as determined by ELISA. (b, c) Cytopathic effects of T‐mB7‐1 and T‐01 in the murine SCC cell lines SCCVII (b) and KLN205‐MUC1 (c). Cell viability is expressed as a percentage relative to mock‐infected controls. Error bars indicate standard deviation (SD). MOI, multiplicity of infection.

Article Snippet: Vero, SCCVII, and SAS cells were seeded in six‐well plates at a density of 2.0 × 10 5 cells per well and incubated for 18 h. The cells were then infected with viruses at a multiplicity of infection of 1.0 for 1 h. After overnight incubation at 34.5°C, the concentration of soluble B7‐1 in the culture supernatant was determined using a mouse CD80 ELISA kit (RayBiotech, GA, USA).

Techniques: Expressing, Activity Assay, Infection, Enzyme-linked Immunosorbent Assay, Standard Deviation