Journal: Signal Transduction and Targeted Therapy

Article Title: Oncolytic adenovirus delivery of neoantigens sensitizes low-mutation tumors to anti-PD-1 therapy and prevents metastasis

doi: 10.1038/s41392-025-02511-5

Figure Lengend Snippet: Neoantigen identification and delivery via oncolytic adenovirus vectors. a Schematic representation of neoantigen identification combining WES and MS. This figure was created via resources from BioRender ( www.biorender.com ). b Design of the neoantigen delivery system. The expression cassette of neoantigen cDNA sequences corresponding to the 29-mer long peptides covering the mutated epitopes was inserted into the E3 region of the survivin promoter (SVP)-regulated oncolytic adenovirus vector Ad5SVPF11 to generate a novel OAV, AdSVP-NAg. C57BL/6 mice inoculated with B16F10 melanoma, MC38 colon carcinoma, or Hep53.4 hepatocellular carcinoma cells were treated with PBS, vector (Ad5SVPF11), SLP, vector combined with SLP, or AdSVP-NAg. Tumor growth curves ( c ) ( n = 6 mice per group), representative images ( d ) of the IFN-γ ELISpot of neoantigen-specific T cells from the spleen in each group and immunofluorescence images ( e ) of tumor tissues. The data are shown as the means ± SDs ( c ) and are representative of two independent experiments ( c – e ). Significance was calculated by two-way ANOVA ( c ). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The formulation of the SLP vaccine was adapted from previous studies, with each dose containing 20 μg of each 29-mer neoantigen peptide in Supplementary Table and 50 μg of poly I:C (MedChemExpress, USA), dissolved in 200 μL of a 10% DMSO/90% PBS solution.

Techniques: Expressing, Plasmid Preparation, Enzyme-linked Immunospot, Immunofluorescence

Journal: Vaccines

Article Title: Immunogenicity of Theileria parva p67C Antigen Delivered via Adjuvanted CoPoP Liposomes in Cattle and Mice

doi: 10.3390/vaccines14050459

Figure Lengend Snippet: p67C-CoPoP liposome nanoparticle characterisation. ( A ) Schematic representation of the generation of CoPoP liposomes with p67C antigen, including the schematic illustrations of the chemical structures of the key adjuvanting molecules. ( B ) Binding capacity of the p67C antigen to CoPoP liposomes after 3 h incubation in the dark, analysed using HisPur™ Ni-NTA Magnetic Beads and assessed by SDS-PAGE. Formulations included: CA (CoPoP only), CQ (CoPoP/QS-21), CP (CoPoP/PHAD), and CPQ (CoPoP/PHAD/QS-21), compared to soluble p67C (PBS). Non-liposome-bound antigen is shown in lanes “B”, while liposome-bound is shown in lanes “S”. The uncropped blots are shown in . ( C ) Binding capacity of fluorophore-labelled p67C-DY490 to CoPoP liposomes (CA, CQ, CP, and CPQ) was measured and compared with p67C bound to AS01, and ( D ) size and ( E ) zeta potential of p67C bound to CoPoP liposomes were measured by DLS.

Article Snippet: Peptide epitope specificity of the immunised bovine sera was evaluated using ten overlapping biotinylated 15-mer peptides to p67C (Mimotopes Pty, Victoria, Australia [ ]), as previously described [ ].

Techniques: Liposomes, Binding Assay, Incubation, Magnetic Beads, SDS Page, Zeta Potential Analyzer

Journal: Vaccines

Article Title: Immunogenicity of Theileria parva p67C Antigen Delivered via Adjuvanted CoPoP Liposomes in Cattle and Mice

doi: 10.3390/vaccines14050459

Figure Lengend Snippet: IgG responses in both cattle and mice immunised with p67C using different CoPoP liposomes. Serum samples from cattle immunised with ( A ) p67C-CA, ( B ) p67C-CP, ( C ) p67C-CQ and ( D ) p67C-CPQ were evaluated using a p67C quantitative ELISA. Kinetics of individual animals are presented in panels ( A – D ), and the mean and standard error of the groups are presented in panel ( E ). Antigen inoculation times are indicated with black arrows in the x-axis. ( F ) Semi-quantification of antibody titres at day 42 in mice immunised with p67C-CPQ or p67C-Alum is presented as the mean of the last positive dilution and standard error per group. Significance is represented by a “*” for p < 0.05.

Article Snippet: Peptide epitope specificity of the immunised bovine sera was evaluated using ten overlapping biotinylated 15-mer peptides to p67C (Mimotopes Pty, Victoria, Australia [ ]), as previously described [ ].

Techniques: Liposomes, Enzyme-linked Immunosorbent Assay

Journal: Vaccines

Article Title: Immunogenicity of Theileria parva p67C Antigen Delivered via Adjuvanted CoPoP Liposomes in Cattle and Mice

doi: 10.3390/vaccines14050459

Figure Lengend Snippet: Cellular response specific to p67C by means of the INFγ-ELISpot assay. ( A ) CD4 + T-cell, ( B ) CD8 + T-cell and ( C ) PBMC responses to 15-mer and 25-mer based on the p67C sequence and p67C protein used as stimuli. Irrelevant peptides and proteins based on the sequence of p67N (N-terminal portion of p67) were used as negative controls. Cells were stimulated for 20 h at 37 °C. Results are presented as the group mean ± standard deviation (SD) of the fold change in the number of spot-forming units compared to the cells stimulated with media only (background). Dotted red lines indicate the cut-off from which the response can be considered different from the background (fold change of 2).

Article Snippet: Peptide epitope specificity of the immunised bovine sera was evaluated using ten overlapping biotinylated 15-mer peptides to p67C (Mimotopes Pty, Victoria, Australia [ ]), as previously described [ ].

Techniques: Enzyme-linked Immunospot, Sequencing, Standard Deviation

Journal: Vaccines

Article Title: Immunogenicity of Theileria parva p67C Antigen Delivered via Adjuvanted CoPoP Liposomes in Cattle and Mice

doi: 10.3390/vaccines14050459

Figure Lengend Snippet: IgG reactivity to p67C 15-mer overlapping peptides. Day 42 sera from cattle immunised with p67C-CoPoP liposomes: Group 3 (p67C-CQ) and Group 4 (p67C-CPQ) were analysed by means of an ELISA assay. Results are presented as a heat map of the relative reactivity compared to the sum of responses against all peptides for each animal.

Article Snippet: Peptide epitope specificity of the immunised bovine sera was evaluated using ten overlapping biotinylated 15-mer peptides to p67C (Mimotopes Pty, Victoria, Australia [ ]), as previously described [ ].

Techniques: Liposomes, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Comprehensive antigen profiling predicts post-surgical neuropathic pain in women treated for breast cancer

doi: 10.1038/s41598-026-41637-6

Figure Lengend Snippet: Characterization of antibody epitope profiles from the serum of patients with CPSNP ( n = 27) and non-CPSNP ( n = 30) by MVA. ( a ) Clinical study design. Samples from 57 patients with breast cancer (BC) diagnosis were analyzed. Each patient had given two samples: one before (timepoint 1, T1) and another after (timepoint 2, T2) surgery. The period between T1 and T2 varied between 4 and 9 years. The control group (CTRL) included serum samples derived from T1 and T2 timepoints from six patients from the same cohort without ICBN injury and without pain. ( b ) Age distribution of the 63 patients at T1 and T2. ( c ) Antibody epitope profiling with MVA. MVA workflow comprised different sequential steps: (i) immunocapture of IgG-phage complexes from a sample using the phage library (random 12-mer peptide M13 phage library); (ii) high-throughput Illumina HiSeq2500 short DNA sequencing of bar-coded phage DNA; (iii) bioinformatical data analysis resulting in antibody epitope identification. The sequenced DNA reads obtained were quality-checked, translated to peptide sequences, and sample- and cohort-specific epitopes from peptide sets were developed by the SPEXS2 pattern search algorithm. Schematic presentation was created with biorender.com ( d ) Highly individual immunoreactive epitope profiles shared similar features in paired sera (T1 and T2) samples, as observed from MVA. The 5000 most IgG-bound (abundant) peptide values (read counts) from each sample were taken into immunoprofile similarity analysis. For all sample pairs, the normalized scalar products of peptide count vectors were calculated for the cosine similarity index (CSI, R package lsa). The CSI values depicted are 0.70 and above (color-coded scale on the right), showing highly correlated immune profiles. Based on CSI analysis, the data of one CPSNP, one non-CPSNP and two CTRL samples were removed from quantitative analyses (Table ). Group sizes: Paired samples n = 59, Unpaired sample pairs = 6844. Sample pairs were combined from timepoint samples of cohort patients: CPSNP n = 26, non-CPSNP n = 29 and CTRL n = 4 (Table ). BC: breast cancer; CPSNP: chronic post-surgical neuropathic pain; CSI: cosine similarity index; CTRL: control; MVA: mimotope variance analysis; NGS: next generation sequencing; NP: neuropathic pain; T1: timepoint 1; T2: timepoint 2.

Article Snippet: In brief, plasma samples ( n = 126, 2 μl of plasma per analysis) were incubated with Escherichia coli M13 phage library displaying random 12-mer peptides (complexity 1 × 10 9 peptides; NEB, E8111L).

Techniques: Biomarker Discovery, Control, Derivative Assay, High Throughput Screening Assay, DNA Sequencing, Next-Generation Sequencing