Journal: Research

Article Title: Inverse Vaccine Nanomimics: In situ Antigen-Specific Immune Tolerance Induction for Durable Cardiac Repair Post-Myocardial Infarction

doi: 10.34133/research.1464

Figure Lengend Snippet: Phenotypic changes and inflammatory factor secretion of T cells stimulated by cardiac lysate antigens (CLAs). (A) Coculture protocol for dendritic cells (DCs) and T cells. The tolerogenic DCs (tDCs) present antigens to T cells; meanwhile, programmed death ligand 1 binds to programmed cell death protein 1 on T cells and inhibits T cell activation, promoting the phenotypic conversion of T cells to regulatory T cells (Tregs). Furthermore, due to the lack of activation signals such as CD80 and CD86, some T cells exhibit a weak activation state after receiving antigen presentation, and under the regulation of key factors such as interleukin-10 (IL-10) and transforming growth factor-β (TGF-β), they undergo phenotypic transition to Tregs. Under restimulation, tDCs phagocytose antigens and present them to Tregs. Tregs are rapidly activated, express high levels of Forkhead box P3 (FOXP3), and directly inhibit T cell activation, while secreting large amounts of anti-inflammatory factors. (B) Flow cytometry (FC) results for T cell phenotype. (C) Quantification of T cell phenotypes by FC. (D) T cell phenotype immunofluorescence staining results, where blue represents cell nuclei, red represents CD127, and green represents CD4. (E) Expression of interferon-γ (IFN-γ), IL-2, IL-10, and TGF-β after coculture of DCs and T cells. (F) Western blot analysis of the activation status of the mechanistic target of rapamycin (mTOR)-p70S6K-S6 pathway after treatment with Gel@PPM, Rapa, inverse vaccine nanomimics (IVNMs), and Gel@IVNMs under CLA stimulation. (G) The percentage of phosphorylation of mTOR, p70S6K, and S6 was statistically analyzed based on the gray values of the western blot results (compared to the unphosphorylated proteins). Data are presented as means ± SD, with individual points representing biological replicates. Statistical significance is indicated as ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For T cell immunofluorescence labeling, nuclei were labeled with 4′,6-diamidino-2-phenylindole (DAPI) (E-CK-A163, Elabscience, China), CD4 with FITC (E-AB-F1097C, Elabscience, China), and CD127 with Alexa Fluor 555 (16-1271-85, eBioscience, USA).

Techniques: Activation Assay, Immunopeptidomics, Flow Cytometry, Immunofluorescence, Staining, Expressing, Western Blot, Analysis, Phospho-proteomics

Journal: Research

Article Title: Inverse Vaccine Nanomimics: In situ Antigen-Specific Immune Tolerance Induction for Durable Cardiac Repair Post-Myocardial Infarction

doi: 10.34133/research.1464

Figure Lengend Snippet: Changes in dendritic cell phenotype and regulatory T cell responses in cardiac-draining lymph nodes after myocardial infarction (MI). (A) Representative flow cytometry plots showing CD11c + MHC-II + dendritic cells and programmed death ligand 1 (PD-L1) expression in the Control, Gel@PPM, Gel@PP@Rapa, Gel@OVA-IVNMs, and Gel@IVNMs groups at 7 and 56 d after MI. (B) Quantification of the proportion of PD-L1 + cells within the CD11c + MHC-II + dendritic cell population at 7 and 56 d. (C) Quantification of CD4 + CD25 + FOXP3 + regulatory T cells (Tregs) in lymph node-derived cells from the 5 treatment groups after 48 h of culture under unstimulated Control, OVA, or cardiac lysate antigen (CLA) stimulation at 7 and 56 d. (D) Representative flow cytometry plots of CD25 and Forkhead box P3 (FOXP3) expression within CD4 + T cells in the Gel@OVA-IVNMs and Gel@IVNMs groups following Control, OVA, or CLA stimulation at 7 and 56 d. Data are presented as means ± SD, with individual points representing biological replicates. Statistical significance is indicated as ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For T cell immunofluorescence labeling, nuclei were labeled with 4′,6-diamidino-2-phenylindole (DAPI) (E-CK-A163, Elabscience, China), CD4 with FITC (E-AB-F1097C, Elabscience, China), and CD127 with Alexa Fluor 555 (16-1271-85, eBioscience, USA).

Techniques: Flow Cytometry, Expressing, Control, Derivative Assay

Journal: Veterinary Medicine and Science

Article Title: Rare Feline Gingival Extramedullary Plasmacytoma: Clinical Application of Electrochemotherapy for Local Control

doi: 10.1002/vms3.71294

Figure Lengend Snippet: Cytologic, histopathologic, immunohistochemical and flow cytometric findings of the gingival mass and regional lymph nodes. (A) Fine‐needle aspiration cytology of the gingival mass showing a low‐cellularity smear composed of a homogeneous population of round cells. The cells exhibit round to oval, occasionally indented and pleomorphic nuclei, with mild to moderate anisocytosis and anisokaryosis. Some cells contain prominent nucleoli, and occasional multinucleated cells are observed. A single neutrophil is present in the upper right field. (B) Histopathologic examination of the gingival mass demonstrating a densely cellular, poorly demarcated, infiltrative round cell neoplasm with frequent mitotic figures. (C and D) Immunohistochemical staining showing strong and diffuse cytoplasmic immunoreactivity of neoplastic cells for MUM‐1, consistent with extramedullary plasmacytoma. (E) Cytologic evaluation of the submandibular lymph node revealed a predominant population of large, atypical round to ovoid cells with marked nuclear atypia, prominent and multiple nucleoli, binucleation and multinucleation, admixed with small to intermediate‐sized lymphocytes, with one abnormal mitotic figure observed in the field, supporting metastatic involvement of the regional lymph node. (F) Flow cytometric immunophenotyping of the lymph node demonstrating heterogeneous expression of CD5 and CD21, with a predominant population of CD5‐ and CD21‐double‐negative cells, supporting the metastatic involvement of extramedullary plasmacytoma in the regional lymph node. Scale bars: A, E = 10 µm; B, D = 50 µm; C = 200 µm.

Article Snippet: For flow cytometric analysis, antibodies against CD21 (B‐cell marker; clone CA2.1D6, RPE‐conjugated, Bio‐Rad Antibodies, MCA1781R), CD5 (pan–T‐cell marker; clone FE1.1B11, FITC‐conjugated, Bio‐Rad Antibodies, MCA2038F), CD4 (helper T‐cell marker; clone VPG34, FITC‐conjugated; Bio‐Rad Antibodies, MCA1346F) and CD8 (cytotoxic T‐cell marker; clone VPG9, RPE‐conjugated; Bio‐Rad Antibodies, MCA1347R) were used.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: bioRxiv

Article Title: Co-targeting KRAS and BCL-2 induces pyroptosis and antitumor tertiary lymphoid structures

doi: 10.64898/2026.10.12.756217

Figure Lengend Snippet: a, Kaplan–Meier survival curves for overall survival (OS) in a published pan-cancer cohort (left) and an immunotherapy cohort (middle), stratified by KRAS mutation and co-mutation subtypes; progression-free survival (PFS) and treatment response in the single-center cohort (right). b, IPS (up) and ESTIMATE score analysis (down) from TCGA (left) and GSE72094 (right) datasets comparing G12D versus non-G12D tumors within STK11-low/co-mutant KL settings. c, Lung weights of Kras^G12D/+;Stk11^f/f (KDL) versus Kras^G12C/+;Stk11^f/f (KCL) mice. d, Single-cell RNA-seq showing proportions of CD8+ T cells, macrophages, and dendritic cells (DCs) in KDL versus KCL tumors. e, f, GSEA of antigen presentation and interferon response pathways in macrophages (e) and DCs (f) from single-cell data. g, Representative IHC images of CD8+ T cell infiltration (left) and quantitative analysis (right) in KL patients with STK11 mutation/low expression versus other subtypes. Data are shown as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Mice were injected intraperitoneally with monoclonal antibodies (200 μg/mouse) targeting CD4 (clone GK1.5, BioXCell BE0003-1), CD8 (clone 53-6.7, BioXCell BE0003-1), or NK cells (clone PK136, BioXCell BE0036) twice weekly.

Techniques: Mutagenesis, Analysis, Single Cell, RNA Sequencing, Immunopeptidomics, Expressing

Journal: bioRxiv

Article Title: Co-targeting KRAS and BCL-2 induces pyroptosis and antitumor tertiary lymphoid structures

doi: 10.64898/2026.10.12.756217

Figure Lengend Snippet: a, b, Antitumor efficacy of combination treatment in KP-1 and FC1242 subcutaneous xenograft models. n = 8 mice per group. c, Lymphoid immune cell composition in KP1 tumors. n = 6 per group. d, Myeloid immune cell composition in KP1 tumors. n = 6 per group. Statistical significance was determined by one-way ANOVA test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Mice were injected intraperitoneally with monoclonal antibodies (200 μg/mouse) targeting CD4 (clone GK1.5, BioXCell BE0003-1), CD8 (clone 53-6.7, BioXCell BE0003-1), or NK cells (clone PK136, BioXCell BE0036) twice weekly.

Techniques: Xenograft Assay, Immune Cell

Journal: bioRxiv

Article Title: Co-targeting KRAS and BCL-2 induces pyroptosis and antitumor tertiary lymphoid structures

doi: 10.64898/2026.10.12.756217

Figure Lengend Snippet: a, Experimental schematic of the in vivo depletion and adoptive transfer study design. b, Tumor growth curves of mice treated with the combination regimen following depletion of B cells, CD4+ T cells, CD8+ T cells, NK cells, or FcyR blockade. n = 5-8 mice per group. c, Multiplex immunofluorescence staining for C3d, IgG, and CK8/18 on tumor sections from vehicle, single-agent, and combination treatment groups. n = 3 per group. d, Quantification of memory B cells in the spleen and tumor-draining lymph nodes from vehicle and combination treatment groups by flow cytometry. n = 4 per group. e, Quantification of long-lived plasma cells in the bone marrow from vehicle and combination treatment groups by flow cytometry. n = 5 per group. f, Serum levels of tumor-specific antibodies measured by flow cytometry in vehicle and combination treatment groups. n = 3-5 per group. g, (Left) Quantification of plasma cells generated from CD19+ splenocytes stimulated with tumor antigens in vitro. (Right) IgG binding to tumor cells detected by flow cytometry after incubation with culture supernatants from vehicle or combination treatment groups. h, Tumor volume curves of naive recipient mice receiving adoptive transfer of B cells or T cells from vehicle- or combination-treated donor mice, followed by tumor inoculation and half-dose RMC-6236 administration. n = 7 mice per group. i, Rechallenge tumor growth in non-CR and treatment-naive mice. n = 8 mice per group. Statistical significance was determined by one-way ANOVA test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Mice were injected intraperitoneally with monoclonal antibodies (200 μg/mouse) targeting CD4 (clone GK1.5, BioXCell BE0003-1), CD8 (clone 53-6.7, BioXCell BE0003-1), or NK cells (clone PK136, BioXCell BE0036) twice weekly.

Techniques: In Vivo, Adoptive Transfer Assay, Multiplex Assay, Immunofluorescence, Staining, Flow Cytometry, Clinical Proteomics, Generated, In Vitro, Binding Assay, Incubation